Network Pharmacology Interpretation of Fuzheng–Jiedu Decoction Against Colorectal Cancer

Hindawi Evidence-Based Complementary and Alternative Medicine Volume 2021, Article ID 4652492, 16 pages https://doi.org/10.1155/2021/4652492

Research Article Network Pharmacology Interpretation of Fuzheng–Jiedu Decoction against Colorectal Cancer

Hongshuo Shi ,1 Sisheng Tian,2 and Hu Tian 3

1College of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, Shandong, China 2School of Management, Shandong University of Traditional Chinese Medicine, Jinan, Shandong, China 3College of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, Shandong, China

Correspondence should be addressed to Hu Tian; [email protected]

Received 7 April 2020; Revised 3 January 2021; Accepted 21 January 2021; Published 20 February 2021

Academic Editor: George B. Lenon

Copyright © 2021 Hongshuo Shi et al. ,is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Introduction. Traditional Chinese medicine (TCM) believes that the pathogenic factors of colorectal cancer (CRC) are “deficiency, dampness, stasis, and toxin,” and Fuzheng–Jiedu Decoction (FJD) can resist these factors. In this study, we want to find out the potential targets and pathways of FJD in the treatment of CRC and also explain from a scientific point of view that FJD multidrug combination can resist “deficiency, dampness, stasis, and toxin.” Methods. We get the composition of FJD from the TCMSP database and get its potential target. We also get the potential target of colorectal cancer according to the OMIM Database, TTD Database, GeneCards Database, CTD Database, DrugBank Database, and DisGeNET Database. Subsequently, PPI analysis, KEGG pathways analysis, and GO biological processes analysis were carried out for the target of FJD in the therapy of colorectal cancer. In addition, we have also built a relevant network diagram. Results. In this study, we identified four core compounds of FJD in the therapy of colorectal cancer, including quercetin, kaempferol, beta-sitosterol, and stigmasterol. At the same time, we also obtained 30 core targets, including STAT3, INS, TP53, VEGFA, AKT1, TNF, IL6, JUN, EGF, CASP3, MAPK3, MAPK1, MAPK8, SRC, IGF1, CCND1, ESR1, EGFR, PTEN, MTOR, FOS, PTGS2, CXCL8, HRAS, CDH1, BCL2L1, FN1, MMP9, ERBB2, and JAK2. FJD treatment of colorectal cancer mainly involves 112 KEGG pathways, including FoxO (hsa04068) signaling pathway, PI3K-Akt (hsa04151) signaling pathway, HIF-1 (hsa04066) signaling pathway, T cell receptor (hsa04660) signaling pathway, and ErbB (hsa04012) signaling pathway. At the same time, 330 GO biological processes were summarized, including cell proliferation, cell apoptosis, angiogenesis, inflammation, and immune. Conclusions. In this study, we found that FJD can regulate cell proliferation, apoptosis, inflammation and immunity, and angiogenesis through PI3K-Akt signaling pathway to play an anti-CRC effect.

1. Introduction while 15 to 25 percent of CRC patients will be found with liver metastasis after radical surgery [3, 4]. At present, the Colorectal cancer (CRC), as the third most common cancer main treatment methods for CRC include radiotherapy and in the world, accounts for the fourth highest cancer mortality chemotherapy, surgery, targeting, and immunotherapy rate in the world [1]. It is a common malignant tumor of the [5, 6]. Because of its limitations, these treatments present the digestive tract. Most patients’ colorectal cancer is caused by whole body toxicity, drug resistance, low selectivity, low their old age and unhealthy lifestyle, and only a few of them efficacy, and other adverse reactions [7]. ,erefore, we want are caused by genetic factors. Other causes of colorectal to find a more effective and less toxic method against CRC cancer may include chronic inflammation of the gut, [8]. overweight, smoking, excessive intake of processed foods, Herbal medicine and traditional medical practice have and inadequate exercise [2]. In the diagnosis, due to the lack been reported as a world upsurge by the World Health of effective clinical screening biomarkers, early symptoms of Organization’s global Atlas of traditional, complementary, CRC are difficult to find. 15 to 25 percent of colorectal cancer and alternative medicine [9]. Traditional Chinese medicine patients have had liver metastasis at the time of detection, (TCM) has been used by the Chinese for thousands of years. 2 Evidence-Based Complementary and Alternative Medicine

It is a part of Oriental traditional medicine. It not only has 2. Materials and Methods the characteristics of good curative effect and small side effects but also has the overall and systematic philosophy 2.1. Collection of Compounds in FJD. ,e compounds of RS, [10]. Chinese herbal medicine treatment can control tumor BX, and TFL in FJD were extracted from the TCM systems growth, improve immune function, regulate tumor micro- pharmacology (TCMSP) database. TCMSP database is one environment, and play a significant role in preventing cancer of the most comprehensive databases of Chinese herbal relapse and transfer. medicine, which is used to find the relationship among ,ere are four main factors in TCM Syndromes of co- herbs, natural compounds, target protein, and diseases [20]. lorectal cancer: “deficiency, dampness, stasis, and toxin” [11]. According to the contemporary Chinese medicine masters, 2.2. Screening of the Active Ingredients in FJD. Although FJD the treatment of tumors should be combined with tonifying is composed of many natural compounds, only a few of them deficiency and resisting evil [12]. In the early stage, we an- have pharmacological effects. ,erefore, it is necessary to alyzed the literature and data mining of the experience of five screen the pharmacokinetics of each compound of Chinese batches of national famous experts in traditional Chinese herbal medicine [21]. Based on the previously reported medicine (500 in each batch, 3000 in total) and three sessions model, we screened various compounds in FJD according to of Chinese Medicine Masters (30 students each, a total of 90 the pharmacokinetic absorption (A), distribution (D), people) in the treatment of CRC. At the same time, we found metabolism (M), and excretion (E) parameters. Based on the the combination of the Fuzheng–Jiedu Decoction (FJD). FJD previous literature and the information in the relevant consists of Panax ginseng C.A. Mey (“RenShen” in Chinese, Chinese herbal medicine database, we selected natural RS), Pinellia ternata (“BanXia” in Chinese, BX), and Smilacis compounds with oral bioavailability (OB) greater or equal to Glabrae Rhizoma (“TuFuLing” in Chinese, TFL). Not only 30 percent and drug sensitivity (DL) greater or equal to 0.18 does the drug combination have a good antitumor effect, but for further analysis [22] (Supplementary file 1). also it can improve the symptoms of CRC such as asthenia and abdominal distension [13]. Based on the theory of TCM, the compatibility of FJD can not only treat “deficiency, 2.3. Get the Target Protein of the Selected Compounds. All the dampness, stasis, and toxin” but also have mild efficacy and active ingredients were input into the TCM systems phar- strong synergistic effect. RS can treat deficiency and improve macology (TCMSP) database to get the known targets. ,e immunity and antitumor, and BX and TFL can treat toxin, 2D structure of the compounds obtained from PubChem stasis, and dampness. Ginsenosides (TGCG) in RS guide was imported into the SwissTargetPrediction database, and a generation cycle retardant and apoptosis in HT-29 cells [14]. threshold (probability >0.6) was set to obtain more credible BX can inhibit the hyperplasia of tumor cells, cut down the targets for each compound. expression of tumor proteins, induce apoptosis of tumor cells, and act on multiple organ tumor cells [15]. Various com- 2.4. Search Targets of CRC. We collected the CRC targets pounds isolated from TFL have been proved to hold up the from six resources: (1) Online Mendelian Inheritance in growth and hyperplasia of CRC cells in varying degrees [16]. Man database (OMIM); (2) ,erapeutic Target Database Although the pharmacological research of RS, BX, TFL, and (TTD); (3) GeneCards Human Gene database (score ≥ 30); their monomers has made some achievements, the traditional (4) Comparative Toxicogenomics Database (CTD); (5) pharmacological evaluation method is difficult to analyze the DrugBank Database; and (6) DisGeNET Database complex mechanism of FJD synergistic treatment of CRC. (Score_gda ≥0.3). ,e official names of the genes (FJD and ,erefore, this study through the network pharmacology CRC) were obtained as UniProt, and the species was selected elaborated its mechanism of action, hoping to provide the as “Homo sapiens.” ,en, the names of the targets were theoretical basis for the compatibility of TCM and provide uniformly converted into gene and UniProt ID [23]. Finally, more methods for the treatment of CRC. we map the target genes of FJD active constituent and CRC Network pharmacology is an effective new strategy in- related target genes, and screen the coincident target genes as tegrating bioinformatics, chemical informatics, network the related targets of FJD treatment for colorectal cancer. biology, traditional pharmacology, and network analysis [17]. Network pharmacology, according to the model of “gene-target-disease,” systematically discusses the relation- 2.5. PPI Network of Target Protein Interaction. ,e target ship between medicine and diseases, which accords with the gene of FJD for CRC was input into the STRING database, holistic view of TCM theory [18]. In order to reveal the and its amplification and prediction were carried out. ,e natural active compounds and potential mechanisms of network diagram of the target protein interaction in vivo was traditional Chinese medicine prescriptions from a system- obtained. We first amplified the target protein of FJD against atic and holistic perspective, network pharmacology pro- CRC with the STRING database and then input it into vides a new method for them [19]. ,e goal of this study was Cytoscape software. We use the “analysis network” tool in to identify the natural compounds of FJD by network Cytoscape to get the protein interaction network to obtain pharmacology and to explore the key target of FJD in the the relevant parameters. Based on the four parameters of treatment of colorectal cancer, so as to comprehend its “Degree,” “BetweennessCentrality,” “ClosenessCentrality,” potential mechanism of action. Our plan is the expression in and “Stress,” we do topology analysis on the PPI network to Figure 1. get hub nodes [24]. Evidence-Based Complementary and Alternative Medicine 3

RS BX TFL OB + DL Colorectal screening cancer

H O O H O O H

O Target ﬁshing H OO H

Target ﬁshing

Common targets collecting

PPI network

KEGG and GO enrichment analysis

Figure 1: Flow chart of this network pharmacology research.

2.6. Enrichment Analysis of GO and KEGG Pathway. GO bioinformatics network platform) database for GO bio- biological process analysis and KEGG pathway analysis are logical process analysis and KEGG pathway analysis and eﬀective methods to explain the role of target genes [25]. identiﬁed GO biological process with P less than or equal We input related hub target genes of FJD treatment CRC to 0.01 and KEGG pathway with P less than or equal to into DAVID 6.8 (high throughput functional labeling 0.01. 4 Evidence-Based Complementary and Alternative Medicine

2.7. Construction of a Network of Herbs, Nature Compounds, 50.41% Targets, and Pathways. On the basis of the compounds, 43.59% 43.07% 43.59% targets, and pathways of FJD in the treatment of colorectal cancer, combined with the functions of Cytoscape 3.7.1 29.70% software, a network model of herbs, ingredients, targets, and 26.83% diseases was constructed. According to this network model, 19.23% 10.57% we made a scientific analysis of the relationship between 10.89% herbs, ingredients, targets, and diseases. We constructed (1) “compound-target” network diagram; (2) “herbal-compound-target” network diagram; (3) PPI network diagram of OB DL OB + DL FJD potential targets for CRC; and (4) “pathway-target” C.A.Mey network diagram. Nodes in the network diagram are linked Panax ginseng Pinellia ternata by edges, and these nodes can represent herbs, compounds, Smilacis Glabrae Rhizoma targets, and KEGG pathways. Figure 2: Column chart of composition screening. 3. Results nodes. ,e obtained targets are divided into two parts, in- 3.1. Preparation by the Screening of the Natural Active In- cluding 468 targets (58 public targets and 410 other human gredients in FJD. According to the TCMSP database, there protein targets), which may represent FJD in in vivo response are 202, 123, and 78 compounds in FJD, respectively. ,ese to CRC. ,ere are nodes 468 and 12359 edges in total. ,en, compounds were screened by OB greater than or equal to 30 the topological characteristics of PPI are analyzed. We use the percent and DL greater than or equal to 0.18. After filtering “analysis network” tool in Cytoscape to get the protein in- and deduplicating values, 46 potential compounds were teraction network to obtain the relevant parameters, and included. ,ere are 22, 13, and 15 compounds in RS, BX, and based on the four parameters of “Degree,” “Betweenness- TFL, respectively. ,e composition screening diagram is Centrality,” “ClosenessCentrality,” and “Stress,” the index shown in Figure 2. Among the 46 selected compounds, above the median value is selected as the key index [26] stigmasterol and beta-sitosterol were found in RS, BX, and (Figure 5) and the core nodes of FJD acting on CRC are TFL. obtained. ,e threshold value of the first screening is “De- gree” ≥37.5, “ClosenessCentrality” ≥0.481692, “Between- 3.2. Study on the Targetof Effective Compounds of FJD. In this nessCentrality” ≥0.000684, and “Stress” ≥4828. ,en, through process, we screened 46 active compounds according to the topological analysis data screening again, 85 hub targets chemical similarity and got 212 related targets, including RS are obtained. ,e criteria for screening were “Degree” ≥62.5, 128, BX 91, and TFL 188. As shown in Figure 3, we have “ClosenessCentrality” ≥0.602967, “BetweennessCentrality” established a “compound-target” network diagram com- ≥0.001387, and “Stress” ≥1172. We got 85 hub nodes and posed of 252 nodes and 842 edges. We use the “analysis established the related network diagram. ,e network dia- network” tool in Cytoscape to get the “Degree” parameter of gram (Figure 6) was composed of 85 nodes and 2698 edges. the network, and we discover that the top four compounds By analyzing the diagram, the top 30 targets are STAT3, INS, of degree are quercetin (degree � 146), kaempferol TP53, VEGFA, AKT1, TNF, IL6, JUN, EGF, CASP3, MAPK3, (degree � 75), beta-sitosterol (degree � 51), and stigmasterol MAPK1, MAPK8, SRC, IGF1, CCND1, ESR1, EGFR, PTEN, (degree � 45) (Table 1). As shown in Figure 4, we also built a MTOR, FOS, PTGS2, CXCL8, HRAS, CDH1, BCL2L1, FN1, “herbal-compound-target” network diagram composed of MMP9, ERBB2, and JAK2. ,ese target genes (Table 2) may 256 nodes and 889 edges. ,rough this diagram, we can be the core targets of FJD in the treatment of CRC. easily observe the relationship between herbs, ingredients, and targets and reveal the potential pharmacological effects of FJD. ,e conclusion of the network graph is consistent 3.4. Enrichment Analysis of KEGG Pathway and GO Biological with the multitarget effect of Chinese herbal medicine and Process. After enrichment analysis of 85 hub targets by the synergistic effect of multidrug compatibility. DAVID v6.8, we got 112 KEGG pathways and 330 GO biological processes according to P < 0.01.

3.3. PPI Network Analysis. ,rough the method of multi- source database integration, the CRC target data in OMIM, 3.4.1. KEGG Pathway Analysis. According to the P value, we TTD, GeneCards, CTD, DrugBank, and DisGeNET data- selected the ﬁrst 20 KEGG (Figure 7) pathways for analysis. bases are integrated. A total of 1018 CRC targets were ob- KEGG pathways, mainly FoxO (hsa04068) signaling pathway, tained according to the screening and removal of duplicate PI3K-Akt (hsa04151) signaling pathway, HIF-1 (hsa04066) values. ,e intersection of FJD and CRC related targets was signaling pathway, T cell receptor (hsa04660) signaling collected and 58 common targets were obtained as the re- pathway, and ErbB (hsa04012) signaling pathway. At the lated targets of FJD in CRC. same time, we have constructed the network diagram between We input 58 target genes of FJD for colorectal cancer into the ﬁrst 20 KEGG pathways and 78 the targets. ,e network the STRING database and got the PPI network diagram of 468 diagram (Figure 8) is composed of 98 nodes and 522 edges. Evidence-Based Complementary and Alternative Medicine 5

CAMK2B ALK AKT1 NEK6PLA2G1B CXCR1 CA5A NEK2 BACE1 MET AXL CSNK2A1 NUAK1 CA9 EGLN1 CYCS KDM4E AKR1C2 AHR XDH CA14 VEGFA ALOX15 AKR1C1 TP53 ALOX12 PKN1 AKR1C4 CDK1 GRK6 CA6 MPO HTR2A HTR2C ADRA1B PNP CA13 ACHE PTPN1 PLK1 AKR1A1 PIK3CG OPRD1 ADRB2 NOX4

CA3 CHRNA2 RXRA ADRA1D TYR GPR35

MMP3 GABRA5 CHRM4 SLC6A4 FLT3 MAPT

MMP13 ADRA2C malkangunin OPRM1 TOP2A PDE3A alexandrin_qt Chrysanthemaxanthin ALOX5

PTK2 Ginsenoside-Rh4_qt ginsenoside rh2 MYLK GABRA2 F7 ESR2 CA7

SRC Panaxadiol Diop APEX1 MAP2 HSD17B2 HTR3A DPP4 CA1 ODC1 JUN Deoxyharringtonine Aposiopolamine ABCC1 Fumarine suchilactone PYGL NOS3 MAPK14 ACACA

PON1 kaempferol Inermin HSD17B1 DAPK1 arachidonate Dianthramine MGAM PTGS2 GSK3B BCL2 CA12 ADORA2A baicalein Frutinone A IL6

CHRM5 CDK2 gondoic acid sitosterol PIK3R1 NR1H3 ESRRA MAPK1

DRD4 NPC1L1 F10 coniferin Girinimbin CHEK1 ABCB1 CTSD 10,13-eicosadienoic diosgenin AURKB CCL2 NCOA2 CYP1B1 PPARG RXRB beta-sitosterol quercetin EGFR beta-D-Ribofuranoside, PLAT CTRB1 Neoastilbin ABCG2 xanthine-9 SCN5A PRSS1 CYP19A1 GJA1 IGHG1 ADORA1 12,13-epoxy-9-hydroxynonadeca-7,10-dienoic Stigmasterol 4,7-Dihydroxy-5-methoxyl-6-methyl-8-formyl-ﬂavan IGF1R astilbin THBD ADRB1 acid PIM1 ADH1C CA4 AVPR2 Cavidine cis-Dihydroquercetin NQO1 naringenin AR CCNA2 PRKACA (3S,6S)-3-(benzyl)-6-(4-hydroxybenzyl)piperazine-2,5-quinone Enhydrin GLO1 KCNMA1 PTGER3

CHRNA7 SYK IL1B ESR1 Baicalin isoengelitin PDE10A SOD1

IFNG MAOA MMP9 COL1A1 CHRM1 24-Ethylcholest-4-en-3-one taxifolin F3 Cycloartenol (-)-taxifolin CALM MAOB (2R,3R)-2-(3,5-dihydroxyphenyl)-3,5,7-trihydroxychroman-4-one MMP2 EGF KCNH2 NR3C2

CACNA1S LTA4H PTPRS POR F2 PGR PDE4D LACTB INSR SULT1E1 CHRM3 NR3C1 HSP90 PLAU AKR1C3 TOP1 DRD1 SRD5A1 TOP2 GABRA1 PTGS1 ADRA2A TNF COL3A1 NOS2 SLC6A2 CA2 KDR AKR1B1 MMP1 RB1 PPARD SLC6A3 SELE HSPA5 CHRM2 VCAM1 PPARA GABRA3 CYP3A4 ACPP ADRA1A CYP1A2 MAPK8 FABP4 IL2 PPP3CA FASN GSTM2 CAT GSTM1 GOT1 HMOX1 GSTP1 CBR1 SHBG HMGCR Figure 3: ,e “compound-target” network diagram of Fuzheng–Jiedu Decoction in the treatment.

Table 1: Compound information sheet. Molecule ID Molecule name Structure OB (%) DL Herb

H O O H O

H Quercetin MOL000098 O O 46.43 0.28 TFL

H O

O H 6 Evidence-Based Complementary and Alternative Medicine

Table 1: Continued. Molecule ID Molecule name Structure OB (%) DL Herb

H OO H O

Kaempferol MOL000422 H 41.88 0.24 RS O O

O H

Beta-sitosterol MOL000358 H 36.91 0.75 RS, BX, and TFL H

HH H O

H Stigmasterol MOL000449 H 43.83 0.76 RS, BX, and TFL H

HH H O

H O

H Diosgenin MOL000546 H 80.88 0.81 TFL O HH

O H

H O H H HO

Ginsenoside rh2 MOL005344 H 36.32 0.56 RS H

O O HO O O H H HO Evidence-Based Complementary and Alternative Medicine 7

Table 1: Continued. Molecule ID Molecule name Structure OB (%) DL Herb

H O O

O H

Baicalein MOL002714 O O 33.52 0.21 BX H

malkangunin alexandrin_qt Chrysanthemaxanthin

Ginsenoside-Rh4_qt ginsenoside rh2

Panaxadiol Diop

Deoxyharringtonine Aposiopolamine

Fumarine suchilactone

kaempferol Inermin arachidonate Dianthramine

AKR1C2 FABP4 MYLK IL2 DRD4 GABRA5 IGF1R PTK2 ESRRA

SYK ADRA1A MMP9 FLT3 ADRB1 CYP1A2 CACNA1S CBR1 TOP2 AURKB LTA4H EGLN1 AKR1C1 AXL CAMK2B baicalein Frutinone A FASN HSD17B2 F3 GABRA2 AKR1C3 ADORA1 gondoic acid sitosterol COL1A1 PTPRS EGF BACE1 MGAM EGFR TOP2A CTSD PTGER3

OPRD1 GSTM1 CAT SELE MPO MMP2 RXRB MET KCNMA1 DAPK1 PPARD AKR1C4 CTRB1 PPP3CA MAPK8

PLAU ALOX5 INSR MAPK1 HSD17B1 MAOB CA3 PLA2G1B PLK1 CA9 PDE4D CA13 CYCS SHBG ADRA2C coniferin Girinimbin RS TFL BX KDR CHRM5 KDM4E MMP1 PON1 GLO1 10,13-eicosadienoic diosgenin AKT1 IL6 NEK2 PLAT PKN1 HTR2C HMGCR CA5A ACACA

CYP19A1 NOX4 HMOX1 CYP3A4 GABRA3 GSTM2 ALK RB1 ADH1C MAPT GRK6 NEK6 PYGL AVPR2 PDE10A

VCAM1 ABCB1 IL1B ABCG2 ADORA2A MMP13 TYR GPR35 THBD NUAK1 NQO1 CALM CHRNA2 beta-sitosterol quercetin beta-D-Ribofuranoside, xanthine-9 Neoastilbin CXCR1 CA6 MAP2 HSPA5 CCL2 PNP GOT1 POR TOP1

AKR1A1 CA1 CSNK2A1 SRD5A1 GJA1 SULT1E1 PIK3R1 PPARA ACPP

12,13-epoxy-9-hydroxynonadeca-7,10-dienoic SRC ODC1 ADRA2A CA14 APEX1 MMP3 COL3A1 Stigmasterol 4,7-Dihydroxy-5-methoxyl-6-methyl-8-formyl-ﬂavan astilbin acid

Cavidine cis-Dihydroquercetin

(3S,6S)-3-(benzyl)-6-(4-hydroxybenzyl)piperazine-2,5-quinone naringenin Enhydrin

Baicalin isoengelitin

24-Ethylcholest-4-en-3-one taxifolin

Cycloartenol (-)-taxifolin (2R,3R)-2-(3,5-dihydroxyphenyl)-3,5,7-trihydroxychroman-4-one

NCOA2 NR1H3 PRKACA NOS3 ALOX12 NR3C1 ESR2 TNF

SLC6A3 MAPK14 PTGS1 SCN5A PDE3A F10 PIM1 AR

ABCC1 ESR1 PGR F7 CHRM1 GSK3B AHR CA2

CCNA2 AKR1B1 CDK1 NOS2 DPP4 BCL2 OPRM1 CA12

NPC1L1 KCNH2 MAOA CHRM3 CA7 JUN ADRA1B LACTB

CDK2 SLC6A2 HSP90 IGHG1 PTPN1 RXRA HTR2A CYP1B1

TP53 CHEK1 PPARG CHRM2 HTR3A PTGS2 CHRNA7 ADRB2

SOD1 GABRA1 SLC6A4 DRD1 VEGFA ACHE IFNG PRSS1

CA4 PIK3CG ALOX15 ADRA1D F2 CHRM4 XDH GSTP1

NR3C2 Figure 4: ,e “herbal-compound-target” network diagram of Fuzheng–Jiedu Decoction in the treatment of colorectal cancer. Green represents herbals. Pink represents the compounds. Blue represents the targets.

MCAT E2F1 ACSS1 CDKN2A APAF1 CTTN TIMP3 ABCB1 STAT5B CAT F10 SLCO1B1HLCS GSTO1 SLC22A8UQCRBPTGES3C19orf48 MMP9 AKR1C2 CALR INSR AKR1D1SRD5A1 MT-CYBSLC2A9 RAD51 CDC25A CDH1 SRD5A2OATHSD17B7HSD3B2PLAUGSTT2BNPAS2 NR0B2 ERBB3 PCOLCEACSS2 SLC22A12 IGF2 CDK6 RHOA DHDH GSTO2 VKORC1UQCRFS1NAT1 CDC25CPPP2CA AKT1 PTGIS CYP3A5NAT2ABCC2 SHC1 COL5A1HSD17B6COX5A NQO1 GNAI2MDM2 NR3C1 CASP8 MAPK9 EPHX1GSTM1FURIN ATMPTPN1ESR2 MDM2 CDK4 COL5A2AGMATF9HSD3B1AKR1C1CYP1A2SERPINC1COMTMRPS7COX5BSPNS2 P4HB TSC2FOXO3 AOC1 GCLMHSD17B1 CYC1 CDC42CD44CDK2 CYP2E1 UBE2I CASP9 PGRMC1 CYP4A11COL6A1MT-CO1F7 ACACASULT2A1 ABCE1 DC≥37.5 KLK3 PIK3R1 SRCMET DC≥62.5 CREBBP PTEN ACAN F5 DNAJC10FABP1GSTA4 AR GGCX AKR1C3CYP7A1CYP2E1LUMNR1I2CYP1A1DNAJB11COX4I1 TMOD2 PIK3CB GRB2 RELA COL3A1STX4 GSTM2PRSS1 NRP2GPX1CD36 ALOX5 IL2 TFPISAT1 RIPK1 ABCB4 GNB1 PLAUR NOS3RELA KRAS IL2 LTB4R2 CYP1B1MMP13DCNCTNND1ABCC1 BC≥0.000684 PECAM1 KDR IGF1R BC≥0.001387 MTOR COL4A1BSGHSD17B2HSP90B1MMP10NR1I3FABP4VTNSERPINE2NR3C2 STAT4 COL18A1MAPK9 IRS1HSP90AA1 STAT3 NQO1 PPARGC1A RHOACDKN1A PTPN11NCOA1 IFNG MMP7 NR0B2AHR E2F4 STAT5AMTORHIF1A SRC TNF PTGESCOL6A3F3CYP3A4CALRPPIGIKBKGKDM1AABCG2TRADD CASP8PTENHRASCTNNB1 CHEK1 ERBB2 PRDX6PDIA3PTGS1C3 P4HB F2 E2F1DNMT1E2F2 CC≥0.481692 C3 CSF2IL1B PIK3R3CHEK2 CC≥0.602967 ADIPOQ KRAS TNFRSF1BGUSB ODC1TGFANFE2L2TIMP2ESR2ADIPOQRXRA NCOR1 CDH1 TP53 FN1 RHOC CATMED1MMP1HGFFASNATF3ABCB1CANXPMAIP1AGRN PGF EP300CYCS PPARG CEBPBGSK3B IGF1 PTGS2 FIGF P2RY4COL6A2TBXAS1MMP3 SERPINE1COL18A1MPOERBB3LEPCREBBPCYCSCOL1A1 MMP8CATTIMP1IGF1CCND1MAPK8AKT1 ADIPOQ IL1R2 ALOX12GGHCXCL8GSTP1ELANEGNAI2TGFB1 ATF4CEBPANCOA3FKBP5HSPA5TOPBP1CLOCKADK ESR1DUSP1PPIG GNG2 CDKN1A SRM NCOA2MMP9MTORNCOA1E2F3RB1 FKBP4TRAF2 Stress≥4828 GRB2 NCOR1 Stress≥1172 PIK3CA ESR1 NOS3 PTGES2KNG1HMOX1PECAM1CCND1 NRP1 TAF1 VEGFA ATF4 MAPK3 RNASE3P2RY12AZU1ALOX5 IGFBP3PLGARCD44CDK2MDM2 FYNFASN IL6MAPK3 NR3C1MMP2 IL1B VEGFA FOXO3TP53 COL1A2 KEAP1STAT4TIMP3 MAPK9 KHDRBS1ARNTLPRKAG1 FN1PTGS2 CCND1 HGF MAP3K7 IL2RBP2RY14ALOX15PTGDSMMP2 IRS1PPARGTNFRSF1AEP300CDKN1ATAB2HSP90AB1ALK EGFR INSCREBBP HSP90AA1SERPINE1 MMP8IL13IGF2IL18CRP IGF1FLT1NR3C1RAD51 CASP2PIK3CBDDX54 STAT1 MAPK1TNF CDK4 GFAP IL4 CTRCSPINK1 IFNGCXCL1JAK2FOSVEGFAIL6EGFHSP90AA1AKT1PTPN11TP53 CHEK1TFDP1BCL2EXO1 TOP2B STAT3JUN LEPERBB2 MED1 OAZ3 NOS2TIMP1PGFFN1JUNSTAT3ATM BIDMCM4MAP1B MMP1IGFBP3 EP300 TGFB1 CYCSPTPN11 IL10 CTNNB1MAPK3ESR1MAPK8EGFRCHEK2 BAD ARMMP9HMOX1KIT CDKN2ACASP3 INS P2RY13IL4 TNFERBB2CASP8SUMO1APAF1CDK4RHOBNEFL PPARG EGF HBEGF PLAURKDRHRASRELAIKBKB KNG1 PLGSERPINE1GSK3BIL4 CYP3A4RB1 HIF1A VEGFCMBPBPI GNG2GRB2 HIF1ACASP3BCL2L1STAT5BCDC42BCL2L11UBE2IBECN1 CDC45 HGF HSP90AB1 IL2RA STAT1 BCL2L1 IL6 JAK3CTSGINS PTENCEBPBMAPK10KRASSRCPXNCDKN2AGSK3BPRKAA1PELP1 TFF1 BCL2L1JAK2 PIK3CA AHR JAK2 IL5IL2RANOS3RHOAGATA3METFOXO3CDK6 BAK1 LPL EGF IL10 OAZ2 IL1BADCY5IL2 PIK3CAKITCDC25C MDM4YES1TDP1 CXCL8 CEBPB KRT1IL1ACRKPIK3R1ARG1IFNGR1SOCS3CSF2IGF1RPTPN1ERN1PLIN1 NCOA3 FOS CASP9TIMP2CYP1A1DNMT1CALM1 IRS1 GABBR1PTGS2 GNB1STAT5ACASP1STAT1SERPINB2CASP9NFATC3PIAS1CALM1MAPK1 COL1A1HSPA5MAPK10MMP3TGFB1SOCS3FLT1 SUMO1 CRP EGFRMAPK1KDR MMP2 IL3 OAZ1RNASE2ARG2 TSC2 CDC25A TP63IL2RGTOP2A CASP1 ARG1 PIK3R1 PRTN3FOXP3KRT2CDH1 RETNKLK3SHC1PIK3CDPDPK1IL6RPIK3R3SOCS1IRS2 CASP6ATR CLSPN IL10 RXRASTAT5BNR1I2 GSTP1 FOS CDC42 FYN INSR PPP2CATOP1AKAP9NPM1NCOA4 CXCL1 NOS2BECN1PPARGC1AMPO CRP GATA3CYP1B1 STAT5A IGF1R HMOX1 FOSB PRKAA2FOXA1PPFIA3PKN1CMA1 MAPK8IP1PRKAR2B ELANEPXN RETN MAPK8 NFATC2 CDH5CRYZPPFIA2FRS3 RPA1BRSK2 RBM28 HSP90B1 CD44 CDK2 SH3KBP1SELP CALM2TNFRSF8NTRK3FOSL2IFNGR2IL1RAPGFAPCTTN PROM1IL6ST SOCS1 LEPHRAS CASP3 CALM3DUSP1DUSP6RICTOR BCAR1 IFNG CTNNB1 JUN PEA15 SYP EML4 TP53BP2 PLCG1DUSP16 NFE2L2 NCOA2 TOP1 CXCL8 PPFIBP1 MARK2 BTBD1 KEAP1 BCL2L11 PECAM1 NESPPFIBP2ALOX15B ACLY LIPE DUSP7DUSP8PTPRS EREG ABCG2 TIMP1 ATF6 KRT10 BRSK1 TNFRSF1AIL13 IL5 RPS6KA1ADORA1 PTPN7 LRRC4B F3 CEBPA PTPRR IL18 INPPL1RAC1 PRKAB1PPFIA4 IL1R1 BTBD2 KIT CBL CSF2 ATM LRFN4MAP2 PPFIA1 F2 IRF1 ZAK

468 nodes and 12359 edges 184 nodes and 6331 edges 85 nodes and 2698 edges Figure 5: ,e process of topological screening for the PPI network. 8 Evidence-Based Complementary and Alternative Medicine

CXCL8 SRC IGF1 MAPK3 ESR1

ERBB2 EGF

MAPK8 EGFR

RELA FOXO3 IL6 HSP90AA1 IL10 MMP9

PECAM1 CDKN2A GSK3B LEP MMP2 MTOR KDR CDH1 MDM2 IL1B CD44 STAT1 PPARG ATM IL2 IFNG SERPINE1 AKT1 NR3C1 IRS1 PTGS2 PIK3CA TIMP1 HIF1A CSF2 NOS3 CAT CDKN1A CREBBP STAT5B ADIPOQ GRB2 CRP IL4 INS KIT CASP3 CEBPB MAPK9 HGF CASP9 CDK4 TGFB1 CYCS PIK3R1 KRAS IGF1R HMOX1 FN1 CTNNB1 JAK2 CASP8 STAT5A RHOA CDK2 PTPN11 AR JUN TP53 EP300 CDC42

TNF PTEN

STAT3 MAPK1

CCND1 HRAS FOS BCL2L1 VEGFA

Figure 6: Construction of protein-protein interaction network expressed by hub target protein. 85 nodes represent 85 proteins, and 2698 edges represent the interaction between 2698 pairs of proteins. ,e node size and color represent the degree, and the data come from the string.

3.4.2. Gene Ontology Analysis. According to the P value, we 4. Discussion selected the first 25 GO biological processes (Figure 9) for analysis, and the top 10 are (GO: 0008284) positive As the third most common cancer, colorectal cancer patients regulation of cell proliferation, (GO: 0043066) negative are less likely to survive for more than five years, still less regulation of apoptotic process, (GO: 0045944) positive than 10% [25, 27]. Chinese herbal medicine has achieved an regulation of transcription from RNA polymerase II ideal curative effect in the treatment of tumors. However, it promoter, (GO: 0010628) positive regulation of gene is difficult to explain the complex mechanism of the tra- expression, (GO: 0042493) response to drug, (GO: ditional method. ,erefore, the integration of network 0045893) positive regulation of transcription, (GO: pharmacology based on big data bioinformatics into the 0045429) positive regulation of nitric oxide biosynthetic molecular mechanism of Chinese herbal medicine in the process, (GO: 0048015) phosphatidylinositol-mediated treatment of diseases is of great significance. signaling, (GO: 0001934) positive regulation of protein According to the network diagram of “compound-tar- phosphorylation, and (GO: 0038128) ERBB2 signaling get,” we got 4 core compounds, namely, quercetin, pathway. kaempferol, baicalein, and beta-sitosterol. Quercetin is a Evidence-Based Complementary and Alternative Medicine 9

Table 2: Information on 30 core targets. Gene Description UniProt Degree Source STAT3 Signal transducer and activator of transcription 3 P40763 84 Predicted INS Insulin P01308 84 Predicted TP53 Cellular tumor antigen p53 P04637 84 Validated VEGFA Vascular endothelial growth factor A P15692 83 Validated AKT1 RAC-alpha serine/threonine-protein kinase P31749 83 Predicted TNF Tumor necrosis factor P01375 82 Validated IL6 Interleukin-6 P05231 82 Validated JUN Transcription factor AP-1 P05412 81 Validated EGF Pro-epidermal growth factor P01133 81 Validated CASP3 Caspase-3 P42574 80 Predicted MAPK3 Mitogen-activated protein kinase 3 P27361 79 Predicted MAPK1 Mitogen-activated protein kinase 1 P28482 79 Validated MAPK8 Mitogen-activated protein kinase 8 P45983 79 Validated SRC proto-oncogene tyrosine-protein kinase Src P12931 78 Predicted IGF1 Insulin-like growth factor I P05019 78 Predicted CCND1 G1/S-specific cyclin D1 P24385 78 Predicted ESR1 Estrogen receptor P03372 77 Validated EGFR Epidermal growth factor receptor P00533 77 Validated PTEN Phosphatase and tensin homolog P60484 76 Predicted MTOR Serine/threonine-protein kinase mTOR P42345 75 Predicted FOS Proto-oncogene c-Fos P01100 74 Predicted PTGS2 Prostaglandin G/H synthase 2 P35354 73 Validated CXCL8 Interleukin-8 P10145 73 Predicted HRAS GTPase HRas P01112 73 Predicted CDH1 Cadherin-1 P12830 73 Predicted BCL2L1 Bcl-2-like protein 1 Q07817 72 Predicted FN1 Fibronectin P02751 71 Predicted MMP9 Matrix metalloproteinase-9 P14780 71 Predicted ERBB2 Receptor tyrosine-protein kinase erbB-2 P04626 70 Predicted JAK2 Tyrosine-protein kinase JAK2 O60674 69 Predicted kind of polyglycol hydroxyflavonoid widely existing in 4.1. Cell Proliferation. PI3K/Akt signaling pathway is critical nature. It has many functions such as antitumor, anti- for the proliferation of colorectal cancer cells. Among them, oxidation, anti-inflammatory, and immune regulation [28]. PI3K/Akt is an important intracellular signaling pathway, In vitro experiments have shown that quercetin used alone which is closely related to the formation and progress of or in combination with other drugs can inhibit the devel- colorectal cancer. It can participate in cell growth, prolif- opment of CRC tumors by blocking the division cycle of eration, differentiation, and migration. PI3K gene activation cancer cells, reducing the biological activity of cancer cells, and PTEN inactivation common in CRC lead to Akt and and regulating certain cancer-related signaling pathways overexpression of downstream targets, including PKC, [29]. Kaempferol exists in propolis, propolis tea, and veg- promotes cell growth and rescues cells from apoptosis. etables. It is a common flavonoid in food. It has the effect of ,erefore, the inhibition of PI3K/Akt has been widely used antiproliferation and inducing apoptosis of human colon to treat CRC [34, 35]. EGF binds with EGFR, thus activating cancer cells [30]. Beta-sitosterol is one of the most abundant the epidermal growth factor receptor, activating PI3K/Akt phytosterols in the diet, comparable to animal cholesterol pathway, and finally promoting tumor proliferation [36, 37]. [31]. It has been reported that ß-sitosterol can significantly Jun transcription factor (JUN) is a key gene of the AP-1 inhibit the progression of DMH-induced CRC in mice [32]. family of transcription factor complexes. MiR-22 is con- Modern research shows that stigmasterol can destroy the sidered to be an important tumor suppressor miRNAs formation of tumor blood vessels in vivo [33]. ,ese [38, 39], which can significantly inhibit the ability of AP-1 to compounds are the material basis of FJD acting on CRC. bind DNA [40]. According to reports, Jun can downregulate According to the analysis of KEGG enrichment, FJD the expression of p53 (TP53), while p53 can activate the treatment of CRC may be achieved through multiple path- expression of miR-22 [41]. Some results show that the ways, the most important of which may be PI3K/Akt single overexpression of proto-oncogenes in fibroblasts makes it pathway. After enrichment analysis of KEGG pathways and easy for c-jun (JUN) to bind to cyclin D1 (CCND1), thereby GO biological process, we found that the potential mecha- activating the expression of cyclin D1 and promoting the cell nism of FJD in the treatment of CRC may be related to its cycle process [42]. Cell division cycle-associated protein 2 participation in cell proliferation, cell apoptosis, inflamma- may target CCND1 by promoting PI3K/Akt pathway, thus tion and immune, and angiogenesis. Next, we will elaborate promoting the abnormal proliferation of colorectal cancer on these four aspects in combination with the core targets. cells [43]. IGF1 (insulin-like growth factor I) is a powerful 10 Evidence-Based Complementary and Alternative Medicine

Top 20 of KEGG enrichment

Non-small cell lung cancer KEGG enrichment. Focal adhesion

Viral carcinogenesis

ErbB signaling pathway

T cell receptor signaling pathway

Colorectal cancer

Chronic myeloid leukemia

Melanoma

Glioma

Endometrial cancer

Bladder cancer p value Pathway 1.230e – 18 HIF−1 signaling pathway 9.225e – 19 Prolactin signaling pathway 6.150e – 19 PI3K−Akt signaling pathway 3.075e – 19

Pancreatic cancer 6.400e – 51

FoxO signaling pathway

Proteoglycans in cancer

Prostate cancer

Hepatitis B

Pathways in cancer

0.1 0.2 0.3 0.4 0.5 Rich factor Gene number 20 40 30 50 Figure 7: KEGG enrichment. mitogenic factor in CRC cells and an important antiapoptotic upregulating DNA repair protein, and maintaining genome factor in CRC cells. IGF1 can bind to its related receptor stability [46, 47]. At the same time, p53 mutants can activate IGF1R to induce PI3K/Akt pathway. As we all know, the liver the Akt signal to promote cancer cell invasion. Integrin and is the most common metastatic site of CRC. ,e source of EGFR depend on the circulation of RCP to participate in this IGFI is the liver. ,erefore, inhibiting IGF1R may be an process [48]. In addition, in many cancers, including co- eﬀective method for the treatment of colorectal cancer [44]. lorectal cancer, the expression of cyclooxygenase-2 (PTGS2) Tumor cell proliferation plays a key role in the occurrence is out of control, while the expression of cyclooxygenase-2 is and development of human malignant tumors. Quercetin is negatively regulated by p53 [49]. As an inhibitor, p53 in- derived from Chinese herbal medicine TFL, which can act on hibits the expression of some antiapoptotic genes, including four targets: JUN, MAPK1, EGF, and EGFR. It may be the main Bcl-2 and promotes apoptosis [50]. Bcl-2 is an apoptotic compound of FJD to inhibit cancer cell proliferation. It has been inhibitor protein, and BCL2L1 belongs to Bcl-2 protein reported that quercetin can inhibit proliferation and induce family [51]. Bax is an apoptotic protein, which can induce apoptosis of SW480, HCT116, HT 29, Caco-2, LoVo, and other the formation of apoptotic bodies and upregulate the ex- CRC cells [45]. Also, baicalein (Table 1) can inhibit MAPK, Akt, pression of Caspase-3 (CASP3). BCL2L1 can steady the or mTOR to hold up the abnormal proliferation of cancer cells. chondriosome localization of Bax and make it lose its biological activity [52]. ,ese proteins are the regulators of apoptosis in PI3K-Akt pathway [53]. It has been shown that 4.2. Cell Apoptosis. Tumor suppressor p53 (TP53) can CXCL8 has a negative correlation with the expression of Bcl- control the progression of cancer by inducing apoptosis, 2-related cell death protein (BAD), thus inhibiting the Evidence-Based Complementary and Alternative Medicine 11

GRB2 KIT IGF1R MDM2 KRAS MAPK9 PIK3R1 HIF1A CREBBP

EP300 HGF

CDK2 STAT1 Prolactin signaling Non-small cell lung pathway Focal adhesion CDK4 cancer CDKN1A HIF-1 signaling Pathways in cancer CYCS pathway GSK3B

Proteoglycans in Viral carcinogenesis RELA cancer FOXO3

HSP90AA1 IL10 PI3K-Akt signaling Bladder cancer pathway AR TP53 IGF1 PTEN CDH1 EGF CAT

TNF MAPK1 FN1 CCND1 CASP3 ErbB signaling PIK3CA Endometrial cancer IRS1 SRC MAPK8 PTGS2 CXCL8 AKT1 pathway FOS JUN IL6 JAK2 ESR1

RHOA MTOR INS STAT3 EGFR HRAS ATM

Hepatitis B MAPK3 BCL2L1 MMP9 ERBB2 VEGFA Pancreatic cancer CASP8 NOS3

T cell receptor CASP9 Glioma IL4 signaling pathway

CDKN2A KDR Melanoma Colorectal cancer

CDC42 FoxO signaling IL2 Prostate cancer Chronic myeloid pathway CTNNB1 leukemia TIMP1

TGFB1 HMOX1

MMP2 SERPINE1 STAT5B IFNG PPARG PTPN11 STAT5A CD44 CSF2

Figure 8: “Target-pathway” network. ,e yellow represents the core targets. ,e blue represents the other targets. ,e orange represents the related pathways. apoptosis of colorectal cancer cells [54]. Besides, mir-204 apoptosis inhibits PI3K/Akt and apoptosis inhibitor Bcl-2, may inhibit the expression of CXCL8 by inhibiting PI3K/ and the accumulation of HIF-1α also has an inhibitory effect Akt/mTOR pathway [55]. Also, the most important way [62]. Diosgenin (Table 1) can upregulate p38 MAPK FOXO participates in apoptosis is PI3K/Akt pathway, which pathway-related proteins, thus promoting TRAIL (a triggers the expression of death receptor ligands such as TNF member of the TNF cytokine family) induced apoptosis [63]. apoptosis ligand and Bcl-2 family members [56]. ,e disorder of apoptosis is directly or indirectly related to the occurrence of tumors [57]. In FJD, the compounds 4.3. Inflammation and Immune. Patients with inflammatory acting on TP53 are quercetin, baicalein, and diosgenin. ,e bowel disease (IBD) including ulcerative colitis have a higher results show that baicalein can inhibit angiogenesis [58] and probability of developing colitis-related CRC [64]. During prevent tumor metastasis [59], and it can control the ab- inflammation, immune cells can lead to the production of normal proliferation and cell cycle of cancer cells by highly genotoxic oxygen/nitrogen-reactive substances, such inhibiting Ezrin and activating p53 pathway-related proteins as IL-6 and TNF-α [6], and form an inflammatory micro- [60]. Besides, a combination of kaempferol and 5-fluoro- environment. ,e production of these inflammatory factors uracil induces apoptosis through PI3K/Akt signaling is mediated through the following signaling pathways, such pathway [61]. Studies show that quercetin-induced as signal transducers and transcription activator 3 (STAT3), 12 Evidence-Based Complementary and Alternative Medicine

Top 25 of GO enrichment Negative regulation of cell proliferation Positive regulation of ERK1 and ERK2 cascade Cellular response to hypoxia Positive regulation of DNA replication Positive regulation of smooth muscle cell proliferation Positive regulation of MAP kinase activity Epidermal growth factor receptor signaling pathway Signal transduction Aging Angiogenesis Positive regulation of peptidyl−tyrosine phosphorylation MAPK cascade Regulation of phosphatidylinositol 3−kinase signaling Positive regulation of sequence−specific GO term DNA binding transcription factor activity p value Response to estradiol 1.0e – 10 ERBB2 signaling pathway 7.5e – 11 Positive regulation of protein phosphorylation 5.0e – 11 Phosphatidylinositol−mediated signaling 2.5e – 11 Positive regulation of nitric oxide biosynthetic process Positive regulation of transcription, DNA−templated Response to drug Positive regulation of gene expression Positive regulation of transcription from RNA polymerase II promoter Negative regulation of apoptotic process Positive regulation of cell proliferation 0.0 0.1 0.2 0.3 Rich factor Gene number 10 25 15 30 20 35 Figure 9: GO biological processes enrichment.

PI3K/AKT, cyclooxygenase-2 (PTGS2), and JAK/STAT. IL- the tumor-bearing host can aggravate the immunosuppres- 6 binds to heterodimer receptors, thereby activating gp130 sion in the tumor microenvironment by inhibiting antitumor on the surface of cell membranes [65], activating and cells including CD8 + T cells, thus leading to the metastasis maintaining the phosphorylation level of STAT3, and and colonization of colon cancer cells [71]. promoting cell proliferation, migration, invasion, and Chronic inflammation is an important factor leading to antiapoptosis [66]. At the same time, TNF-α plays a bio- CRC. TNF gene is regulated by quercetin, kaempferol, logical role in tumor cells through the STAT3 signaling astilbin, and ginsenoside rh2 in FJD and IL6 gene is regulated pathway, so the STAT3 signaling pathway is a bridge be- by quercetin. Quercetin has a strong anti-inflammatory effect. tween inflammation and tumor [67]. PTGS2 encodes in- It can reduce some special inflammatory factors including ducible prostaglandin synthase cyclooxygenase-2, which is an TNF-α, by preventing the phosphorylation and degradation important rate-limiting enzyme produced by prostaglandins of IκB in the process of inflammation [72]. Studies have and has a key significance for malignant tumors. Activation of shown that ginsenoside rh2 (Table 1) upregulates the se- the PI3K/Akt signaling pathway can upregulate PTGS2 [68], cretion levels of TNF-α and IL-6 in the serum of colon cancer which has been proved to be involved in the production of mice, relieves tumor-related immunosuppression, and pro- inflammatory prostaglandins through stimulation events and longs the survival of colon cancer mice [73]. in various biological processes such as tumor cell proliferation, angiogenesis, and invasion [69]. Tcell mediated acquired immunity can effectively promote the process of tumor re- 4.4. Angiogenesis. VEGFA can induce angiogenesis by gression. Tumor cells affect immune cells through the STAT3 promoting the abnormal proliferation and migration of signaling pathway and participate in the immune escape of vascular endothelial cells, which is conducive to the growth, tumor cells. ,erefore, STAT3 has become an important invasion, and metastasis of tumor [74]. Hypoxia is the protein in immunotherapy research [70]. IL-6 produced by feature of the tumor microenvironment. Cells under hypoxia Evidence-Based Complementary and Alternative Medicine 13 can produce hypoxia inducible factor-1 α (HIF-1 α), which pathways (DAVID Database, https://david.ncifcrf.gov/) can lead to the enhancement of tumor cell invasion and analysis, and GO biological processes (DAVID Database, metastasis [35]. Under hypoxia state, VEGF enhanced ex- https://david.ncifcrf.gov/) analysis were carried out for the pression and even the initial starting factor of angiogenesis target of FJD in the therapy of colorectal cancer. in tumor tissue, while HIF-1α played a “bridge” role in the process from hypoxia to VEGF increased expression [75]. Conflicts of Interest Under the condition of hypoxia, it has also been found that HIF-1α can also combine with the hypoxia response element ,e authors declare no conflicts of interest. (HRE) of hypoxia response gene, promote the transcription of the downstream target gene to enter the nucleus, promote Authors’ Contributions Akt activation, activate PI3K/Akt signal transduction pathway, inhibit endothelial cell apoptosis, and promote Hongshuo Shi was responsible for the experimental oper- angiogenesis [76]. ation and thesis writing. Sisheng Tian was responsible for the As we all know, angiogenesis is a necessary condition revision of the final version. Hu Tian proposed research and a key step for tumor growth and metastasis. ,erefore, ideas and designed research plans. the inhibition of tumor angiogenesis can block the occurrence, development, invasion, and metastasis of tumors to a Acknowledgments certain extent [77]. In FJD, the main compounds of genes that act on VEGFA are baicalein, diosgenin, and quercetin. ,is project was supported by the Shandong Traditional Quercetin can inhibit tumor angiogenesis by inhibiting Chinese Medicine Development Project (Z135080000022) VEGFR2. Also, some studies have shown that kaempferol and the AcShandong Traditional Chinese Medicine Inher- can inhibit the HIF-1α and VEGFR2 in endothelial cells itance and Development Project (Z135080000022), Network through PI3K/Akt/mTOR signals, thus showing its anti- Pharmacology Interpretation of Fuzheng–Jiedu Decoction angiogenesis ability [78]. against Colorectal Cancer.

5. Conclusion Supplementary Materials Traditional Chinese medicine has been passed down for Supplementary File 1 includes natural compounds con- thousands of years in China. ,e idea of Chinese herbal tained in the Fuzheng–Jiedu Decoction (FJD), and the medicine treating tumors is the combination of tonifying ADME parameter is oral bioavailability (OB) ≥ 30% and deficiency and resisting evil. Based on the combination of drug-likeness (DL) ≥ 0.18. Table (A) in Supplementary File 1 modern medicine, a lot of experience in treating malignant indicates the compounds in RS; Table (B) in Supplementary tumors has been summarized. In this study, we first rec- File 1 indicates the compounds in BX; and Table (C) in ognized from the perspective of network pharmacology that Supplementary File 1 indicates the compounds in TFL. FJD regulates cell proliferation, apoptosis, inflammation, (Supplementary Materials) immunity, and angiogenesis through the PI3K-Akt signaling pathway. In addition, we also provide a scientific explanation References for FJD in treating “deficiency, dampness, stasis, and toxin,” and we hope to open a window for understanding the [1] M. Arnold, M. S. Sierra, M. Laversanne, I. Soerjomataram, complex mechanism of action of Chinese herbal medicine A. Jemal, and F. Bray, “Global patterns and trends in colo- and the essence of more concepts of traditional Chinese rectal cancer incidence and mortality,” Gut, vol. 66, no. 4, medicine. pp. 683–691, 2016. [2] O. K. D. Stephen, “Diet, microorganisms and their metabo- lites, and colon cancer,” Nature Reviews Gastroenterology & Data Availability Hepatology, vol. 13, no. 12, pp. 691–706, 2016. [3] E. J. Kuipers, W. M. Grady, D. Lieberman et al., “Colorectal ,e data used to support the findings of this study are cancer,” Nature Reviews Disease Primers, vol. 1, Article ID available from the corresponding author upon request. ,e 15065, 2015. authors got the composition of FJD from the TCMSP da- [4] H. Zhui, “,e progress of TCM Research on liver metastasis of tabase (http://tcmspw.com/tcmspsearch.php) and its po- colorectal cancer,” Hunan Journal of Traditional Chinese tential target from the TCMSP database (http://tcmspw. Medicine, vol. 35, no. 12, pp. 129–133, 2019. com/tcmspsearch.php) and Swiss TargetPrediction data- [5] L. Faugeras, A. Dili, A. Druez, B. Krug, C. Decoster, and base (http://www.swisstargetprediction.ch/). ,ey also got L. D’Hondt, “Treatment options for metastatic colorectal the potential target of colorectal cancer according to the cancer in patients with liver dysfunction due to malignancy,” Critical Reviews in Oncology/Hematology, vol. 115, pp. 59–66, OMIM Database (http://www.omim.org/), TTD Database 2017. (http://bidd.nus.edu.sg/group/cjttd/), GeneCards Database, [6] R. Francescone, V. Hou, and S. I. Grivennikov, “Cytokines, CTD Database (http://ctdbase.org/), DrugBank Database IBD, and colitis-associated cancer,” Inflammatory Bowel (https://www.drugbank.ca/), and DisGeNET Database Diseases, vol. 21, no. 2, pp. 409–418, 2015. (http://www.disgenet.org/home/). Subsequently, PPI anal- [7] F. Geng, Z. Wang, H. Yin, J. Yu, and B. Cao, “Molecular ysis (STRING Database, https://string-db.org/), KEGG targeted drugs and treatment of colorectal cancer: recent 14 Evidence-Based Complementary and Alternative Medicine

progress and future perspectives,” Cancer Biotherapy and Ethnopharmacology, vol. 176, no. Complete, pp. 402–412, Radiopharmaceuticals, vol. 32, no. 5, pp. 149–160, 2017. 2015. [8] B. Gong, Y. Kao, C. Zhang, H. Zhao, F. Sun, and Z. Gong, [23] W. Wang, T. Liu, L. Yang et al., “Study on the multi-targets “Exploring the pharmacological mechanism of the herb pair mechanism of triphala on cardio-cerebral vascular diseases “HuangLian-GanJiang” against colorectal cancer based on based on network pharmacology,” Biomedicine & Pharma- network pharmacology,” Evidence-based Complementary and cotherapy, vol. 116, 2019. Alternative Medicine, vol. 2019, Article ID 2735050, 12 pages, [24] S.-Y. Xiang, J. Zhao, Y. Lu et al., “Network pharmacology- 2019. based identification for therapeutic mechanism of Ling-Gui- [9] G. Bodeker and C.-K. Ong, WHO Global Atlas of Traditio- Zhu-Gan decoction in the metabolic syndrome induced by nal,Complementary and Alternative Medicine, World Health antipsychotic drugs,” Computers in Biology and Medicine, Organization, Geneva, Switzerland, 2005. vol. 110, p. 1, 2019. [10] G. Yu, W. Wang, X. Wang et al., “Network pharmacology- [25] R. Siegel, D. Naishadham, and A. Jemal, “Cancer statistics,” A based strategy to investigate pharmacological mechanisms of Cancer Journal for Clinicians, vol. 63, no. 1, pp. 11–30, 2013. Zuojinwan for treatment of gastritis,” BMC Complementary [26] Y. Wan, L. Xu, Z. Liu et al., “Utilising network pharmacology and Alternative Medicine, vol. 18, no. 1, p. 292, 2018. to explore the underlying mechanism of Wumei Pill in [11] D. Li, W. Wang, L. Xiang et al., “,e type of Traditional treating pancreatic neoplasms,” BMC Complementary and Chinese Medicine syndrome predicts prognosis and che- Alternative Medicine, vol. 19, no. 1, 2019. motherapeutic outcomes in colorectal cancer,” European [27] J. Chen, C. Li, Y. Zhu et al., “Integrating GO and KEGG terms Journal of Integrative Medicine, vol. 33, 2020. to characterize and predict acute myeloid leukemia-related [12] T. Hu and S. Tian, “Analysis of the experience of the first genes,” Hematology, vol. 20, no. 6, pp. 336–342, 2015. national medical master in the treatment of malignant tu- [28] S. G. Darband, M. Kaviani, B. Yousefi et al., “Quercetin: a mors,” Shi Zhen National Medicine, vol. 30, no. 1, pp. 193-194, functional dietary flavonoid with potential chemo-preventive 2019. properties in colorectal cancer,” Journal of Cellular Physiology, [13] T. Hu, Periodical Literature Analysis of Contemporary Famous vol. 233, no. 9, pp. 6544–6560, 2018. Chinese Medicine Experts in Diagnosis and Treatment of [29] P. Poyil, B. Amit, S. Young-Ok et al., “Quercetin inhibits Colorectal Cancer and Summary of Tutor Experience, Shan- angiogenesis mediated human prostate tumor growth by dong University of traditional Chinese medicine, Jinan, targeting VEGFR- 2 regulated AKT/mTOR/P70S6K signaling China, 2020. pathways,” PLoS One, vol. 7, no. 10, Article ID e47516, 2012. [14] T. Li, W. Sun, X Dong et al., “Total ginsenosides of Chinese [30] S. Yao, X. Wang, C. Li, T. Zhao, H. Jin, and W. Fang, ginseng induces cell cycle arrest and apoptosis in colorectal “Kaempferol inhibits cell proliferation and glycolysis in carcinoma HT-29 cells,” Oncology Letters, vol. 16, no. 4, esophagus squamous cell carcinoma via targeting EGFR p. 4640, 2018. signaling pathway,” Tumor Biology, vol. 37, no. 8, [15] X. Dong, W. Xu, J. Shen, and H. Sun, “Research and appli- pp. 10247–10256, 2016. cation of Pinellia Ternata in the treatment of malignant tu- [31] A. B. Awad and C. S. Fink, “Phytosterols as anticancer dietary mors,” Journal of Traditional Chinese Medicine, vol. 47, no. 4, components: evidence and mechanism of action,” ?e Journal pp. 106–108, 2019. of Nutrition, vol. 130, no. 9, pp. 2127–2130, 2000. [16] S. Hua, Y. Zhang, J. Liu et al., “Ethnomedicine, phyto- [32] M. S. Bin Sayeed and S. S. Ameen, “Beta-sitosterol: a chemistry and pharmacology of smilax glabra: an important promising but orphan nutraceutical to fight against cancer,” traditional Chinese medicine,” ?e American Journal of Chinese Medicine, vol. 46, pp. 261–297, 2018. Nutrition and Cancer, vol. 67, no. 8, pp. 1216–1222, 2015. [17] S. I. Berger and R. Iyengar, “Network analyses in systems [33] T. Kangsamaksin, S. Chaithongyot, C. Wootthichairangsan pharmacology,” Bioinformatics, vol. 25, no. 19, pp. 2466–2472, et al., “Lupeol and stigmasterol suppress tumor angiogenesis 2009. and inhibit cholangiocarcinoma growth in mice via down- [18] D. Bu, Z. Su, J. Zou, M. Meng, and C. Wang, “Study of the regulation of tumor necrosis factor-α,” PLoS One, vol. 12, mechanism underlying therapeutic effect of compound Article ID e0189628, 2017. Longmaining on myocardial infarction using a network [34] C. P. R. Xavier, C. F. Lima, A. Preto, R. Seruca, M. Fernandes- pharmacology-based approach,” Biomedicine & Pharmaco- Ferreira, and C. Pereira-Wilson, “Luteolin, quercetin and therapy, vol. 118, 2019. ursolic acid are potent inhibitors of proliferation and inducers [19] W. Song, S. Ni, Y. Fu, and Y. Wang, “Uncovering the of apoptosis in both KRAS and BRAF mutated human co- mechanism of Maxing Ganshi Decoction on asthma from a lorectal cancer cells,” Cancer Letters, vol. 281, no. 2, systematic perspective: a network pharmacology study,” pp. 162–170, 2009. Scientific Reports, vol. 8, no. 1, 2018. [35] Y. Kitajima, T. Ide, T. Ohtsuka, and K. Miyazaki, “Induction [20] J. Ru, P. Li, J. Wang et al., “TCMSP: a database of systems of hepatocyte growth factor activator gene expression under pharmacology for drug discovery from herbal medicines,” hypoxia activates the hepatocyte growth factor/c-Met system Journal of Cheminformatics, vol. 6, no. 1, p. 13, 2014. via hypoxia inducible factor-1 in pancreatic cancer,” Cancer [21] C. Ma, C. Zhai, X. Tian et al., “A systems pharmacology-based Science, vol. 99, no. 7, pp. 1341–1347, 2008. study of the molecular mechanisms of san cao decoction for [36] M. F. Rimawi, P. B. Shetty, H. L. Weiss et al., “Epidermal treating hypertension,” Evidence-based Complementary and growth factor receptor expression in breast cancer association Alternative Medicine, vol. 2019, Article ID 3171420, 10 pages, with biologic phenotype and clinical outcomes,” Cancer, 2019. vol. 116, no. 5, pp. 1234–1242, 2010. [22] F. Tang, Q. Tang, Y. Tian, Q. Fan, Y. Huang, and X. Tan, [37] J. A. Engelman, J. Luo, and L. C. Cantley, “,e evolution of “Network pharmacology-based prediction of the active in- phosphatidylinositol 3-kinases as regulators of growth and gredients and potential targets of Mahuang Fuzi Xixin de- metabolism,” Nature Reviews Genetics, vol. 7, no. 8, coction for application to allergic rhinitis,” Journal of pp. 606–619, 2006. Evidence-Based Complementary and Alternative Medicine 15

[38] J. Xiong, “Emerging roles of MicroRNA-22 in human disease [54] J. Li, Q. Liu, X. Huang et al., “Transcriptional profiling reveals and normal physiology,” Current Molecular Medicine, vol. 12, the regulatory role of CXCL8 in promoting colorectal cancer,” no. 3, pp. 247–258, 2012. Frontiers in Genetics, vol. 10, 2019. [39] W. Yuan, Y. Li, M. Ding, H. Zhang, X. Xu, and J. Tang, [55] F. Shuai, B. Wang, and S. Dong, “MicroRNA-204 inhibits the “Molecular mechanisms and clinical applications of miR-22 growth and motility of colorectal cancer cells by down- in regulating malignant progression in human cancer,” In- regulation of CXCL8,” Oncology Research, vol. 26, no. 8, ternational Journal of Oncology, vol. 50, pp. 345–355, 2017. p. 1295, 2018. [40] J. Yang, L. Chen, J. Ding et al., “Cardioprotective effect of [56] M. Farhan, H. Wang, U. Gaur, P. J. Little, J. Xu, and W. Zheng, miRNA-22 on hypoxia/reoxygenation induced car- “FOXO signaling pathways as therapeutic targets in cancer,” diomyocyte injury in neonatal rats,” Gene, vol. 579, no. 1, International Journal of Biological Sciences, vol. 13, no. 7, pp. 17–22, 2015. p. 815, 2017. [41] Y. Liu, X. Chen, R. Cheng et al., “,e Jun/miR-22/HuR [57] Y. Wang, C. Chao, Y. Chun, H. Li, and Y. Yang, “Apoptosis of regulatory axis contributes to tumourigenesis in colorectal human colorectal cancer cells induced by 6-gingerol and its cancer,” Molecular Cancer, vol. 17, no. 1, p. 11, 2018. effect on Bax, BCL2, Caspase3 and PARP1 gene expression,” [42] C. Zhang, C. Chang, H. Gao, Q. Wang, F. Zhang, and C. Xu, Modern Food Technology, vol. 33, no. 11, pp. 7–15, 2017. “MiR-429 regulates rat liver regeneration and hepatocyte [58] Z. He, B. Li, G. O. Rankin, Y. Rojanasakul, and Y. C. Chen, proliferation by targeting JUN/MYC/BCL2/CCND1 signaling “Selecting bioactive phenolic compounds as potential agents pathway,” Cellular Signalling, vol. 50, 2018. to inhibit proliferation and VEGF expression in human [43] Y. Feng, W. Qian, Y. Zhang et al., “CDCA2 promotes the ovarian cancer cells,” Oncology Letters, vol. 9, no. 3, proliferation of colorectal cancer cells by activating the AKT/ pp. 1444–1450, 2015. CCND1 pathway in vitro and in vivo,” BMC Cancer, vol. 19, [59] R. R. Ramjiawan, A. W. Griffioen, and D. G. Duda, “Anti- no. 1, 2019. angiogenesis for cancer revisited: is there a role for combi- [44] X. L. Chao, L. L. Wang, R. Liu, Y. Li, and X. J. Zhou, “As- nations with immunotherapy?” Angiogenesis, vol. 20, no. 2, sociation between CA repeat polymorphism in IGF1 gene pp. 185–204, 2017. promoter and colorectal cancer risk in a native Chinese [60] Z. Chen, R. Hou, S. Gao, D. Song, and F. Ye, “Baicalein population,” Neoplasma, vol. 66, no. 6, 2019. inhibits proliferation activity of human colorectal cancer cells [45] X. Qiu and C. Ma, “Research progress of quercetin against HCT116 through downregulation of Ezrin,” Cellular Physi- colon cancer,” Chinese Journal of Traditional Chinese Medi- ology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology, vol. 49, cine, vol. 33, no. 10, pp. 2333–2335, 2015. no. 5, 2018. [46] B. Jean-Christophe, S. Sylvanie, and M. P. Khoury, [61] Q. Li, L. Wei, L. Shan, Y. Chen, J. Lin, and J. Peng, “Synergistic “Uncovering the role of p53 splice variants in human ma- effect of kaempferol and 5-fluorouracil on the growth of lignancy: a clinical perspective,” Onco Targets and ?erapy, colorectal cancer cells by regulating the PI3K/Akt signaling vol. 7, pp. 57–68, 2013. pathway,” Molecular Medicine Reports, vol. 20, pp. 728–734, [47] A. Ventura, D. G. Kirsch, M. E. Mclaughlin et al., “Restoration 2019. of p53 function leads to tumour regression in vivo,” Nature, [62] L. B. Si, M. Z. Zhang, Q Han et al., “Sensitization of keloid vol. 445, no. 7128, pp. 661–665, 2007. fibroblasts by quercetin through the PI3K/Akt pathway is [48] H. Li, J. Zhang, J. H. M. Tong et al., “Targeting the oncogenic dependent on regulation of HIF-1α,” American Journal of p53 mutants in colorectal cancer and other solid tumors,” Translational Research, vol. 10, no. 12, p. 4223, 2018. International Journal of Molecular Sciences, vol. 20, no. 23, [63] C. Lepage, D. Y. Leger,´ J. Bertrand et al., “Diosgenin induces 2019. death receptor-5 through activation of p38 pathway and [49] A. K. Maurya and M. Vinayak, “Anticarcinogenic action of promotes TRAIL-induced apoptosis in colon cancer cells,” quercetin by downregulation of phosphatidylinositol 3-kinase Cancer Letters, vol. 301, no. 2, pp. 0–202, 2011. (PI3K) and protein kinase C (PKC) via induction of p53 in [64] C. Lucas, N. Barnich, and H. T. T. Nguyen, “Microbiota, hepatocellular carcinoma (HepG2) cell line,” Molecular Bi- inflammation and colorectal cancer,” International Journal of ology Reports, vol. 42, no. 9, pp. 1419–1429, 2015. Molecular Sciences, vol. 18, no. 6, p. 1310, 2017. [50] S. N. D. C. Pereira, M. C. Lima, T. B. Boquimpani et al., [65] Y. Yu, W. Wang, S. Zhai, S. Dang, and M. Sun, “IL6 gene “Lycopene extracts from different tomato-based food prod- polymorphisms and susceptibility to colorectal cancer: a ucts induce apoptosis in cultured human primary prostate meta-analysis and review,” Molecular Biology Reports, vol. 39, cancer cells and regulate TP53, Bax and bcl-2 transcript ex- no. 8, pp. 8457–8463, 2012. pression,” Asian Pacific Journal of Cancer Prevention : APJCP, [66] S. Doumas, J. C. Paterson, P. M. Norris et al., “Fractalkine vol. 18, no. 2, 2017. (CX3CL1) and fractalkine receptor (CX3CR1) in squamous [51] Z. Yang, Y. An, N. Wang, X. Dong, and H. Kang, “LINC02595 cell carcinoma of the tongue: markers of nerve invasion?” Oral promotes tumor progression in colorectal cancer by inhib- and Maxillofacial Surgery, vol. 19, no. 1, pp. 61–64, 2015. iting miR-203b-3p activity and facilitating BCL2L1 expres- [67] R. Rosell, J. Bertran-Alamillo, M. A. Molina, and M. Taron, sion,” Journal of Cellular Physiology, vol. 235, pp. 7449–7464, “IL-6/gp130/STAT3 signaling axis in cancer and the presence 2020. of in-frame gp130 somatic deletions in inflammatory hepa- [52] H. Zhang, Y. Zong, G. Qiu et al., “Silencing Lin28 promotes tocellular tumors,” Future Oncology, vol. 5, no. 3, pp. 305–308, apoptosis in colorectal cancer cells by upregulating let-7c 2009. targeting of antiapoptotic BCL2L1,” Molecular Medicine [68] Z. Cao, L.-Z. Liu, D. A. Dixon, J. Z. Zheng, B. Chandran, and Reports, vol. 17, no. 4, 2018. B.-H. Jiang, “Insulin-like growth factor-I induces cyclo- [53] V. Duronio, “,e life of a cell: apoptosis regulation by the oxygenase-2 expression via PI3K, MAPK and PKC signaling PI3K/PKB pathway,” Biochemical Journal, vol. 415, no. 3, pathways in human ovarian cancer cells☆,” Cellular Signal- pp. 333–344, 2008. ling, vol. 19, no. 7, pp. 1542–1553, 2007. 16 Evidence-Based Complementary and Alternative Medicine

[69] C. S. Williams, M. Mann, and R. N. Dubois, “,e role of cyclooxygenases in inﬂammation, cancer, and development,” Oncogene, vol. 18, no. 55, pp. 7908–7916, 1999. [70] J. Wang, F. Huang, C. Jiang, and P. Chi, “Silencing signal transducer and activator of transcription 3 (STAT3) and use of anti-programmed cell death-ligand 1 (PD-L1) antibody induces immune response and anti-tumor activity,” Medical Science Monitor, vol. 26, Article ID e915854, 2020. [71] Y. Toyoshima, H. Kitamura, H Xiang et al., “IL6 modulates the immune status of the tumor microenvironment to facilitate metastatic colonization of colorectal cancer cells,” Cancer Immunology Research, vol. 7, no. 12, p. 1944, 2019. [72] A. Kordkheyli Vahid, K. Tarsi Abbas, M. Mohammad et al., “Eﬀects of quercetin on microRNAs: a mechanistic review,” Journal of Cellular Biochemistry, vol. 120, no. 8, pp. 12141– 12155, 2019. [73] J. Wang, Y. Chen, C. Dai, Y. Shang, and J. Xie, “Ginsenoside Rh2 alleviates tumor-associated depression in a mouse model of colorectal carcinoma,” American Journal of Translational Research, vol. 8, no. 5, pp. 2189–2195, 2016. [74] L. Zhou, D.-S. Wang, Q.-J. Li, W. Sun, Y. Zhang, and K.-F. Dou, “Downregulation of the Notch signaling pathway inhibits hepatocellular carcinoma cell invasion by inactivation of matrix metalloproteinase-2 and -9 and vascular endothelial growth factor,” Oncology Reports, vol. 28, no. 3, pp. 874–882, 2012. [75] W. Li, M. Petrimpol, K. D. Molle, M. N. Hall, E. J. Battegay, and R. Humar, “Hypoxia-induced endothelial proliferation requires both mTORC1 and mTORC2,” Circulation Research, vol. 100, no. 1, p. 79, 2007. [76] V. V. Polosukhin, J. M. Cates, W. E. Lawson et al., “Hypoxia- inducible factor-1 signalling promotes goblet cell hyperplasia in airway epithelium,” ?e Journal of Pathology, vol. 224, no. 2, pp. 203–211, 2011. [77] H. S. Choi, K. Lee, M. K. Kim et al., “DSGOST inhibits tumor growth by blocking VEGF/VEGFR2-activated angiogenesis,” Oncotarget, vol. 7, no. 16, pp. 21775–21785, 2016. [78] Gi D. Kim, “Kaempferol inhibits angiogenesis by suppressing HIF-1α and VEGFR2 activation via ERK/p38 MAPK and PI3K/Akt/mTOR signaling pathways in endothelial cells,” Preventive Nutrition and Food Science, vol. 22, no. 4, 2017.