Research Article Antiproliferative Activity, Proapoptotic Effect, and Cell Cycle Arrest in Human Cancer Cells of Some Marine Natural Product Extract
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Hindawi Oxidative Medicine and Cellular Longevity Volume 2020, Article ID 7948705, 12 pages https://doi.org/10.1155/2020/7948705 Research Article Antiproliferative Activity, Proapoptotic Effect, and Cell Cycle Arrest in Human Cancer Cells of Some Marine Natural Product Extract Hong Cui ,1 Mansour A. E. Bashar ,2 Islam Rady ,2,3 Hussein A. El-Naggar ,2 Lamiaa M. Abd El-Maoula ,4 and Ahmed B. M. Mehany 2 1Department of Hepatobiliary and Pancreatic Surgery, Tumor Hospital of Zhengzhou University, Zhengzhou, China 2Zoology Department, Faculty of Science, Al-Azhar University, Cairo, Egypt 3Masonic Cancer Center, University of Minnesota, Minnesota, USA 4Nutrition and Food Science Department, Faculty of Home Economics, Al-Azhar University, Egypt Correspondence should be addressed to Ahmed B. M. Mehany; [email protected] Received 27 June 2020; Revised 27 October 2020; Accepted 10 November 2020; Published 26 November 2020 Academic Editor: Fabiana Morroni Copyright © 2020 Hong Cui et al. This 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. Bioactive constituents of numerous marine organisms have been investigated recently for their preclinical and clinical anticancer activity. Three marine organisms: black-spotted sea cucumber: Pearsonothuria graeffei (Pg), lollyfish: Holothuria atra (Ha), and sea hare: Aplysia dactylomela (Ad), were collected during winter 2019 from Gulf of Aqaba, Red Sea, Egypt, and macerated with ethanol into three different extracts: PgE, HaE, and AdE, where each was in vitro assessed for its antiproliferative and proapoptotic properties on HepG2, HCT-116, and MCF-7 cancer cells. PgE dose-dependently inhibited the growth of HepG2, HCT-116 MCF-7 μ , and cells within IC50 values 16.22, 13.34, and 18.09 g/mL, respectively, while the IC50 values for the E μ E antiproliferative activity of Ha were 12.48, 10.45, and 10.36 g/mL, respectively, and the IC50 values of Ad were 6.51, 5.33, μ HepG2 and 6.87 g/mL, respectively. All extracts were found to induce G0/G1 cell cycle arrest for cells side by side with their inhibition of CDK2 on all three cell lines while all extracts were also showed to induce apoptosis in HepG2 cell line at pre-G1 phase supplemented by their anticancer activity via proapoptotic protein Bax, caspase-3, and cleavage PARP increase, and antiapoptotic protein Bcl-2 downturn. Moreover, necrosis has been relatively noticed in HepG2 cell line as an additional anticancer activity for each extract. Our data introduced three ethanolic marine extracts as natural chemotherapeutic agents to be further developed for cancer control. 1. Introduction cancer therapy [3]. Natural products have been studied in a variety of models of cancer where the results have been Cancer remains the second causative agent of deaths world- extremely promising [4]. wide [1, 2]. Annually, there are more than 10 million new Water bodies are considered a source rich in bioactive cancer patients with over 6 million correlated deaths compounds which add suitability progress and serve as a accounting for roughly 12% of worldwide mortality [3]. chemical shield against and sometimes protect from other Therefore, there is an urgent need to determine the way of creatures. These bioactive compounds have biological treatment of this ailment where the current strategy in treat- properties such as antioxidant, antibacterial, and antitumor ment are mainly radiation-based therapy and chemotherapy [6, 7]. Marine fauna such as sponges, mollusks, echinoderms, [4]. However, this strategy is associated with toxic effects ascidians, and corals are taxonomically different and have a such as causing hair loss and other serious adverse effects variety of pharmacological activities that give us a tremen- [5]. Nowadays, the utilization of complementary and alterna- dous research opportunity attraction for the discovery of tive medicines is more popular as a promising strategy for novel anticancer therapeutics [8]. Researchers have been 2 Oxidative Medicine and Cellular Longevity looking for marine derivative products 50 years ago, whereas Table 1: Cell viability of PgE, HaE, and AdE on HePG2, HCT-116, in the last few decades, about 3000 new compounds have and MCF-7 cell lines. been isolated from various marine sources and assessed for μ their anticancer efficacy [9]. In recent years, many marine Cell line viability Concentration ( g/mL) (%) 6.25 12.5 25 50 100 IC anticancer bioactive compounds have been isolated, charac- 50 terized, identified, and preclinically assessed, and they are HePG2 70.19 62.36 41.91 26.81 18.34 16.22 under clinical trials for human use [7]. Recently, marine nat- PgE HCT-116 72.37 55.29 37.66 26.45 15.25 13.34 ural products are considered the most interesting source of MCF-7 74.42 64.46 45.37 31.51 20.48 18.09 bioactive compounds and therapeutic agents [7]. During HePG2 65.81 50.74 39.94 25.64 14.37 12.48 the last decades, marine sources are chemically mediated HaE HCT-116 63.44 48.38 38.74 23.29 12.37 10.45 so-called secondary metabolites or natural products [10– MCF-7 66.61 51.84 40.36 30.19 12.55 10.36 12]. In the beginning, secondary metabolites have been iden- tified as metabolic waste products or remains of primary HePG2 50.11 39.47 27.87 16.62 8.58 6.51 metabolic overflow without any obvious biological function, AdE HCT-116 49.78 37.26 25.69 14.27 6.16 5.33 but now, there is no doubt that these substances are playing MCF-7 50.53 40.09 30.36 18.78 8.33 6.87 a fundamental role in ecology and reproduction [13], while primary metabolites such as carbohydrates, amino acids, and fatty acids are chemically almost found in most organ- HepG2 cell line was selected to study the cell cycle analysis isms differ from secondary metabolites which are higher in (Figures 2 and 3). HepG2 cell accumulation was noticed at chemical diversity associated with limited phylogenetic dis- the G0/G1 phase; the percentages were 57.59, 61.12, and tribution, whereas the major structural classes of natural 62.18% upon treating cells with PgE, HaE, and AdE, products include alkaloids, terpenes, polyketides, peptides, respectively. This cell cycle analysis outcome is supported and shikimate-derived metabolites [14]. Marine chemical by a significant inhibition of cell cycle regulatory protein ecology has been known as a kind of young field due to the CDK2 (Figure 4). recent investigations in the last 25 years for the natural com- pounds mediating interactions between marine invertebrates 2.2. Apoptosis and Necrosis. Each extract of Pg, Ha,orAd compared to natural products isolated from terrestrial plants induced apoptosis in HepG2 cell line at the pre-G1 phase and microorganisms since the 1930s, and this is partly due to (Figure 4). In addition, each extract increased the level of the recent development of underwater technology and new the proapoptotic protein Bax and caspase-3 and decreased establishment of the ecologically relevant bioassays [15]. the level of the antiapoptotic protein Bcl-2 on HepG2 can- Besides the nutritional values of sea cucumber, there are ther- cer cell line (Figure 5). Also, each extract induced apopto- apeutic medicinal benefits that can be linked to the presence sis via cleavage PARP increase in HepG2, HCT-116, and of bioactive compounds which generated a lot of unique bio- MCF-7 cancer cells (Figure 6). Annexin V/propidium logical and pharmacological activities such as antiangio- iodide double staining (PI) assay was also used to investi- genic, anticoagulant, antihypertension, anti-inflammatory, gate the mode of induced HepG2 cell death caused by antimicrobial, antioxidant, antithrombotic, antitumor, and each extract treatment (Figure 7). The total percent of cell wound healing [16–18]. A lot of bioactive compounds death is represented in Figure 8. So, necrosis has been rel- have been extracted, characterized, and purified from atively noticed in the percentage of cell death that has many animals, for instance, tunicates, sponges, soft corals, caused in HepG2 cell line as an additional anticancer bryozoans, cephalopods, and echinoderms [19–22]. Also, activity for each extract. among marine organisms, there are important commercial In summary, it seems that PgE, HaE,orAdE each pro- groups such as marine holothurians which are spiny duced marked induction apoptosis and antiproliferative skinned echinoderm invertebrates [23]. Therefore, it is activity on HepG2, HCT-116, and MCF-7 cancer cells via possible to isolate a lot of bioactive compounds from multiple pathways (Figure 9). marine fauna that may contribute to better health and serve as novel medication [24]. Here, an in vitro evalua- tion of the anticancer effects of three marine invertebrate 3. Material and Methods crude extracts is presented. 3.1. Sampling and Identification of Marine Specimens. The 2. Results marine organisms were collected via scuba diving from different sites at the Gulf of Aqaba, Egypt, during winter 2.1. Antiproliferative Activity. All extracts inhibited the cell 2019. The taxonomy details were studied, and the voucher growth, each in all three cell lines in a dose-dependent man- specimen was deposited at Marine Laboratory, Depart- ner with a remarkable cell viability decrease associated with ment of Zoology, Faculty of Science, Al-Azhar University, fi cytotoxicity increase (Table 1 and Figure 1). The IC50 values Egypt, with a registration number of MZ1005. Identi ca- for PgE, HaE, and AdE on HePG2 were 16.22, 12.48, and tion of samples was carried out according to Ruggiero 6.51 μg/mL, respectively, while on HCT-116 were 13.34, et al. [25] as well as consultation of marine scientists 10.45, and 5.33 μg/mL, respectively, whereas on MCF-7 were through the World Wide Web as a confirmation for the 18.09, 10.36, and 6.87 μg/mL, respectively.