DOCSLIB.ORG

Colon Carcinoma Cells Harboring PIK3CA Mutations Display Resistance to Growth Factor Deprivation Induced Apoptosis

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Colon Carcinoma Cells Harboring PIK3CA Mutations Display Resistance to Growth Factor Deprivation Induced Apoptosis 1143 Colon carcinoma cells harboring PIK3CA mutations display resistance to growth factor deprivation induced apoptosis Jing Wang,1 Karen Kuropatwinski,1 Jennie Hauser1 resulted in resistance to GFDS-induced apoptosis, where- Michael R.Rossi, 2 Yunfei Zhou,1 Alexis Conway,1 as transfection of wild-type PIK3CA or empty vector had Julie L.C. Kan,3 Neil W.Gibson, 3 little effect. Taken together, our studies show that mutant James K.V. Willson,4 John K.Cowell, 2 PIK3CA increases the capacity for proliferation and and Michael G.Brattain 1 survival under environmental stresses, such as GFDS while also imparting greater dependency on the PI3K Departments of 1Pharmacology and Therapeutics and pathway for proliferation and survival. [Mol Cancer Ther 2Cancer Genetics, Roswell Park Center Institute, Elm and Carlton 2007;6(3):1143–50] Streets, Buffalo, New York; 3OSI Pharmaceuticals, Broadhollow Bioscience Park, Farmingdale, New York; and 4University of Texas Southwestern Medical Center at Dallas, Dallas, Texas Introduction Phosphatidylinositol 3-kinase (PI3K) is a lipid kinase that catalyzes the transfer of the g-phosphate from ATP to the Abstract D3 position of the phosphatidylinositol, generating phos- A PIK3CA, encoding the p110 catalytic subunit of phos- phatidylinositol 3-phosphate, phosphatidylinositol 3,4- phatidylinositol 3-kinase (PI3K), is mutated in a variety of bisphosphate and phosphatidylinositol 3,4,5-trisphosphate human cancers. We screened the colon cancer cell lines (1). Class I PI3Ks are heterodimers consisting of two sub- previously established in our laboratory for PIK3CA units: an adaptor/regulatory subunit and a catalytic sub- mutations and found that four of them harbored gain of unit (p110). The regulatory subunit is a 50-kDa or 85-kDa function mutations. We have now compared a panel of protein that is tightly associated with the p110 catalytic mutant and wild-type cell lines for cell proliferation and subunit. PI3Ks are activated through receptor tyrosine survival in response to stress. There was little difference in kinases or through G protein–coupled receptors. Class IA PI3K activity between mutant PIK3CA-bearing cells PI3Ks are very diverse in mammals; they have three (mutant cells) and wild-type PIK3CA-bearing cells (wild- catalytic p110 isoforms (p110a, h, and y), each encoded by type cells) under optimal growth conditions. However, the a separate gene, and seven adaptor proteins generated by mutant cells showed constitutive PI3K activity during expression and alternative splicing of three different genes growth factor deprivation stress (GFDS), whereas PI3K (p85a, h, and p55g; ref. 2). activity decayed rapidly in the wild-type cells. Importantly, PI3K activation has been shown to have important roles constitutively active PI3K rendered the mutant cells in sustaining processes important to malignancy, including resistant to GFDS-induced apoptosis relative to the wild- cell proliferation, adhesion, survival, and motility. There type cells, indicating a biological advantage under stress are several downstream effectors of PI3K, including PDK1, conditions that is imparted by the mutant enzymes. Rac, p70S6K, certain isoforms of protein kinase C, and Compared with the wild-type cells, the mutant cells were AKT, which is most relevant to cell survival (1). Phosphor- hypersensitive to the apoptosis induced by the PI3K ylation of phosphatidylinositol-4,5-bisphosphate by PI3K inhibitor LY294002. In addition, PIK3CA small interfering generates phosphatidylinositol 3,4,5-trisphosphate, which RNA significantly decreased DNA synthesis and/or in- then recruits PDK1 to the cell membrane. PDK1 subse- duced apoptosis in the mutant cells but not in the wild- quently phosphorylates and activates AKT. Activated AKT type cells. Furthermore, ecotopic expression of a mutant can then phosphorylate a variety of substrates, such as Bad, PIK3CA in a nontumorigenic PIK3CA wild-type cell line FKHR, caspase-9, and GSK3, that participate in a variety of cellular processes, including cell survival and proliferation. Phosphorylation of Bad and caspase-9 by AKT inhibits the ability of these proteins to induce apoptosis. Therefore, the Received 9/8/06; revised 12/21/06; accepted 1/31/07. PI3K pathway has been shown to inhibit apoptotic Grant support: NIH grants CA 34432, CA54807, and CA 16056. processes and has been linked to inappropriate cell survival The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked by malignant cells in response to stress (3, 4). advertisement in accordance with 18 U.S.C. Section 1734 solely to Overexpression of AKT has an antiapoptotic effect in indicate this fact. many cell types, resulting in a delay of cell death during Requests for reprints: Michael G. Brattain, Department of Pharmacology stress. AKT was found to be amplified in human breast and Therapeutics, Roswell Park Center Institute, Elm and Carlton Streets, Buffalo, NY 14263. Phone: 716-845-3044; Fax: 716-845-8857. (5, 6), ovarian (7), pancreatic (8), and prostate cancer (9), E-mail: [email protected] suggesting the specific involvement of AKT in the onset Copyright C 2007 American Association for Cancer Research. and/or propagation of cancer. In addition, the genomic doi:10.1158/1535-7163.MCT-06-0555 locus encoding the p110a catalytic subunit PIK3CA was Mol Cancer Ther 2007;6(3). March 2007 Downloaded from mct.aacrjournals.org on September 25, 2021. © 2007 American Association for Cancer Research. 1144 Mutant PI3-Kinase Signaling in GFDS-induced Apoptosis found to be amplified in a high percentage of various from PIK3CA haplo-insufficient cells. It has been shown that cancers. PIK3CA gene amplification has been shown to be expression of mutant PI3K enhances lipid kinase activity associated with the progression of several types of tumors and induces constitutive activation of AKT signaling and to an invasive phenotype (10–12). Recent studies have oncogenic transformation in chicken embryo fibroblasts, shown that PIK3CA is mutated in approximately one-third as well as in NIH3T3 cells (17, 18). Because environmental of 234 colorectal cancers but only in 2 of 76 adenomas (13). restriction on growth seems to be common in solid tumors It is also mutated at a high frequency in other cancers, (20), the ability of malignant cells to withstand these stresses including breast, ovarian, and hepatocellular carcinomas is considered a key factor in tumor development and (14–16), making it one of the most common mutations progression. Our finding that the differential cell survival described to date in human cancers. Expression of one of characteristics between the mutant and wild-type cells the PIK3CA mutants in NIH3T3 cells conferred significant- under stress conditions, in which the wild-type PI3K activity ly more lipid kinase activity compared with the wild-type is more severely down-regulated, provides an explanation protein, suggesting the gain-of-function nature of the for the irrelevance of the wild-type PI3K activity in terms of mutations (13). In addition, some of the most frequent the advantage provided by the mutant enzyme. We also mutants of PIK3CA showed transforming activity with found that ecotopic expression of the mutant PI3K in the high efficiency in chicken embryo fibroblasts, as well as in wild-type PI3K-bearing colon cancer cells led to increased NIH3T3 cells (17, 18). Analogy to gain-of-function epider- resistance to GFDS-induced apoptosis. This result indicates mal growth factor receptor (EGFR) mutants in lung cancers that gain-of-function mutant PIK3CA may contribute to the suggests that these PIK3CA mutations may be associated ability to withstand the environmental stresses that are with ‘‘oncogenic addiction’’ and are, therefore, hypersen- important in tumor development and/or progression. sitive to PI3K inhibition. Thus, mutant PI3K may be a promising target for PI3K small molecule inhibitors. Exploration of the roles of mutant PI3K in cancer growth, Materials and Methods survival, and metastasis will help unveil the mechanisms of Cell Culture and Reagents malignant progression and facilitate development of drugs Human colon carcinoma cells used in this study were for cancer treatment. established in vitro from primary tumors, as described This study compares a panel of unmodified PIK3CA previously (21). HCT116, FET, GEO, CBS, and RCA were mutant and wild-type colon cell lines for proliferation and cultured in SM medium [McCoy’s 5A serum-free medium survival under stress condition. We found that mutant (Sigma, St. Louis, MO) with pyruvate, vitamins, amino PIK3CA-bearing cells were more resistant to growth acids, and antibiotics] supplemented with 10 ng/mL factor deprivation stress (GFDS)–induced apoptosis than epidermal growth factor, 20 Ag/mL insulin, and 4 Ag/mL the wild-type PIK3CA-bearing cells, probably due to the transferrin, as described previously (22). RKO and TENN constitutively active PI3K and its downstream effectors. In were cultured in SM medium supplemented with 10% fetal addition, the mutant cells were hypersensitive to a potent bovine serum. Vaco481 were cultured in MEM basal PI3K inhibitor, LY294002, as reflected by significantly medium (MEM medium with glutamine and amino acids) increased apoptosis compared with the wild-type cells. supplemented with 100 Ag/mL insulin, 20 Ag/mL trans- PIK3CA small interfering RNA dramatically decreased ferrin, 8.6 ng/mL selenium, and 2% fetal bovine serum. DNA synthesis and induced
Load more

Recommended publications
  • Potential Dual Role of Activating Transcription Factor 3 in Colorectal Cancer
    ANTICANCER RESEARCH 36: 509-516 (2016) Potential Dual Role of Activating Transcription Factor 3 in Colorectal Cancer XIAOJING JIANG*, KUI-JIN KIM*, TAEKYU HA and SEONG-HO LEE Department of Nutrition and Food Science, College of Agriculture and Natural Resources, University of Maryland, College Park, MD, U.S.A. Abstract. Background/Aim: Activating transcription factor leucine zipper (bZip) DNA binding domain (3). Basal 3 (ATF3) is a member of the ATF/CREB transcription factor expression of ATF3 is low in normal cells and can be family and has been proposed as a molecular target for induced by various stress stimuli and signals that damage cancer therapy. The present study was undertaken in order cells or tissues (4). These stress signals include hypoxia, to investigate whether ATF3 influences cancer-related anoxia, carcinogens, DNA damage, UV exposure and phenotypes in colorectal cancer. Materials and Methods: radiation (5). In addition, ATF3 is responsible for adaptation ATF3 was overexpressed in human colorectal cancer cells to different extra- and intra-cellular stimuli (6, 7). and the effects of ATF3 on apoptosis, cell cycle, cell In previous studies, we found that ATF3 could be a molecular migration and epithelial mesenchymal transition (EMT) were target of many anticancer compounds such as PI3K inhibitor investigated. B-cell lymphoma-2 (Bcl-2) promoter was cloned (8), epicatechin gallate (9), indole-3-carbinol (10), conjugated and used for luciferase assay in cells transfected with control linoleic acid (11), and tolfenamic acid (12), that mediate or ATF3 expression vector. Results: ATF3 down-regulated the compounds-stimulated apoptosis (9-11). However, the role of expression of Bcl-2 and promoter activity of the Bcl-2 gene.
    [Show full text]
  • Thymoquinone Induces Apoptosis and DNA Damage in 5-Fluorouracil-Resistant Colorectal Cancer Stem/Progenitor Cells
    www.oncotarget.com Oncotarget, 2020, Vol. 11, (No. 31), pp: 2959-2972 Research Paper Thymoquinone induces apoptosis and DNA damage in 5-Fluorouracil-resistant colorectal cancer stem/progenitor cells Farah Ballout1, Alissar Monzer1, Maamoun Fatfat1, Hala El Ouweini1, Miran A. Jaffa2, Rana Abdel-Samad3, Nadine Darwiche3, Wassim Abou-Kheir4 and Hala Gali-Muhtasib1,4 1Department of Biology, American University of Beirut, Lebanon 2Department of Epidemiology and Population Health, American University of Beirut, Lebanon 3Department of Biochemistry and Molecular Genetics, American University of Beirut, Lebanon 4Center for Drug Discovery and Department of Anatomy, Cell Biology and Physiological Sciences, American University of Beirut, Lebanon Correspondence to: Hala Gali-Muhtasib, email: [email protected] Wassim Abou-Kheir, email: [email protected] Keywords: thymoquinone; colorectal cancer stem cells; 5-Fluorouracil resistance; colonospheres; apoptosis Received: October 08, 2019 Accepted: December 16, 2019 Published: August 04, 2020 Copyright: Ballout et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT The high recurrence rates of colorectal cancer have been associated with a small population of cancer stem cells (CSCs) that are resistant to the standard chemotherapeutic drug, 5-fluorouracil (5FU). Thymoquinone (TQ) has shown promising antitumor properties on numerous cancer systems both in vitro and in vivo; however, its effect on colorectal CSCs is poorly established. Here, we investigated TQ’s potential to target CSCs in a three-dimensional (3D) sphere-formation assay enriched for a population of colorectal cancer stem/progenitor cells.
    [Show full text]
  • Targeting the Active B-Catenin Pathway to Treat Cancer Cells
    2861 Targeting the active B-catenin pathway to treat cancer cells Hadas Dvory-Sobol,1,2 Eyal Sagiv,1,2 PUMA, whereas that of cells with low levels of B-catenin Diana Kazanov,1 Avri Ben-Ze’ev,3 and signaling was not. Growth inhibition was associated with Nadir Arber1,2 induction of apoptosis. Chemotherapy synergistically enhanced the effect of AdTOP-PUMA. A combination of 1Integrated Cancer Prevention Center, Tel Aviv Sourasky the adenovirus system with standard therapy may 2 Medical Center; Sackler School of Medicine, Tel-Aviv University, improve the efficacy and reduce the toxicity of therapy Tel-Aviv, Israel; 3Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel in humans. [Mol Cancer Ther 2006;5(11):2861–71] Abstract Introduction The adenomatous polyposis coli or b-catenin genes are h-Catenin is a multifunctional protein serving as a major frequently mutated in colorectal cancer cells, resulting in structural component of cell-to-cell adherens junctions. In oncogenic activation of B-catenin signaling. We tried to addition, it also acts as an important signaling molecule in establish in vitro and in vivo models for selectively the Wnt pathway that plays a key role in embryogenesis killing human cancer cells with an activated B-catenin/ and tumorigenesis (1–3). T-cell factor (Tcf) pathway. We used a recombinant In the absence of Wnt signaling, the cytoplasmic level of adenovirus that carries a lethal gene [p53-up-regulated h-catenin is kept low through interaction with a protein modulator of apoptosis (PUMA)] under the control of a complex [containing GSK3h-glycogen synthase kinase 3h, B-catenin/Tcf–responsive promoter (AdTOP-PUMA) to axin and adenomatous polyposis coli (APC)] that can selectively target human colorectal cancer cells phosphorylate h-catenin and target it to ubiquitin-mediated (SW480, HCT116, DLD-1, and LS174T), hepatocellular proteasomal degradation (4).
    [Show full text]
  • Establishment and Validation of Human Colon Cancer HCT116 Wip1 FLAG Knock-In Cell Line
    Open Access Annals of Mutagenesis Research Article Establishment and Validation of Human Colon Cancer HCT116 wip1 FLAG Knock-in Cell Line Cui Ying-Yu1,2,3,4* 1Department of Regenerative Medicine, Tongji University Abstract School of Medicine, China A HCT116 cell line with FLAG Knock-in in the 5’ terminus of its wip1 2Institute of Medical Genetics, Tongji University School of gene was established by genome editing via Adeno-Associated Virus (AAV)- Medicine, China mediated Homologous Recombination (HR) “knock-in” approach and identified 3Key Laboratory of Arrhythmias of the Ministry of by PCR, sequencing and western blot. The corresponding AAV-HR gene Education of China, Tongji University, China targeting can facilitate many protein functional analyses, including western blot, 4Department of Oncology, Georgetown University immunoprecipitation, and protein screening etc. This human somatic cell gene Medical Center, USA targeting technique provides a general solution not only for the study of proteins *Corresponding author: Cui Ying-Yu, Department of without ideal antibodies, but also for screening potential proteins interacting with Regenerative Medicine, Institute of Medical Genetics, Key targeted protein in human somatic cells. Laboratory of Arrhythmias, Tongji University School of Keywords: Genome editing; Gene knock-in; Human somatic cell gene Medicine, Shanghai 200092, China targeting; Adeno-associated virus mediated homologous recombination (AAV- Received: March 27, 2017; Accepted: May 22, 2017; HR); HCT116 cell line; wip1 gene Published: May 29, 2017 Introduction Presently, epitope-tagged expression vectors have been widely used to study gene function [9,10]. However, this kind of transgenes Knock-outs and knock-ins by homologous recombination generally lack features present in endogenous genes such as a natural are powerful genome editing techniques to definitively assess gene promoter, introns and 5’- and 3’- untranslated regions, all of which function.
    [Show full text]
  • Status of P53 in Human Cancer Cells Does Not Predict Efficacy of CHK1
    Oncogene (2010) 29, 6149–6159 & 2010 Macmillan Publishers Limited All rights reserved 0950-9232/10 www.nature.com/onc ORIGINAL ARTICLE Status of p53 in human cancer cells does not predict efficacy of CHK1 kinase inhibitors combined with chemotherapeutic agents S Zenvirt1, N Kravchenko-Balasha1 and A Levitzki Unit of Cellular Signaling, Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel DNA damage checkpoints cause cell cycle arrest, allowing Millar, 2006). An important regulator of this response DNA repair before resumption of the cell cycle. These is the tumor suppressor p53. In response to DNA checkpoints can be activated through several signaling damage, p53 is activated and stabilized by the Ser/Thr pathways. Checkpoint activators include p53, checkpoint kinases Ataxia-Telangiectasia mutated and Ataxia- kinase 1 (CHK1), checkpoint kinase 2 and/or MAPKAP Telangiectasia- and RAD3-related (Oren, 2003; Meek, kinase 2 (MK2). Many cancer cells lack p53 activity and, 2004; Harris and Levine, 2005). The activated p53 therefore, depend on alternative checkpoint activators to protein stimulates the expression of p21, which inhibits arrest the cell cycle following DNA damage. Inhibition of the cyclin-dependent kinases, resulting in cell cycle arrest these pathways is expected to specifically sensitize these at both the G1-to-S (G1/S) and the G2-to-M (G2/M) p53-deficient cells to DNA damage caused by chemother- transitions (Vogelstein et al., 2000). In the absence of apy. Using isogenic p53-proficient and p53-deficient cancer p53, the G1/S checkpoint cannot be properly activated. cell lines, we show that inactivation of CHK1, but not Intra-S and G2/M checkpoints can be activated through MK2, abrogates cell cycle arrest following chemotherapy, the Ser/Thr kinases checkpoint kinase 1 (CHK1), specifically in p53-deficient cells.
    [Show full text]
  • Rapid Inhibition of Cancer Cell Growth Induced by Lentiviral Expression of Mutant-Template Telomerase RNA Constructs and Anti-Telomerase Sirna
    UCSF UC San Francisco Previously Published Works Title Rapid inhibition of cancer cell growth induced by lentiviral expression of mutant-template telomerase RNA constructs and anti-telomerase siRNA Permalink https://escholarship.org/uc/item/85n1b5hz Journal Cancer Research, 64 Authors Li, Shang Rosenberg, Jonathan Donjacour, A. A. et al. Publication Date 2004 Peer reviewed eScholarship.org Powered by the California Digital Library University of California [CANCER RESEARCH 64, 4833–4840, July 15, 2004] Rapid Inhibition of Cancer Cell Growth Induced by Lentiviral Delivery and Expression of Mutant-Template Telomerase RNA and Anti-telomerase Short-Interfering RNA Shang Li,1 Jonathan E. Rosenberg,1 Annemarie A. Donjacour,2 Inna L. Botchkina,1 Yun Kit Hom,2 Gerald R. Cunha,2 and Elizabeth H. Blackburn1 Departments of 1Biochemistry and Biophysics and 2Anatomy, University of California at San Francisco, San Francisco, California ABSTRACT stem cells, and germ cells (12–16). In contrast, telomerase activity is highly elevated in 85–90% of human cancers and Ͼ70% of immor- In human cancers, telomeres are commonly maintained by elevated talized human cell lines (14, 17). This is consistent with up-regulated levels of the ribonucleoprotein enzyme telomerase, which contains an telomerase conferring a strong selective advantage for continued intrinsic templating RNA moiety (human telomerase RNA; hTER) and the core protein (human telomerase reverse transcriptase). We developed growth of malignant cells (18). Although cells without telomerase can a lentiviral system for efficient overexpression of mutant-template human be tumorigenic in mouse xenografts, they cannot be immortalized telomerase RNA (MT-hTer) to add mutant DNA to telomeres in cancer (19).
    [Show full text]
  • FTO Downregulation Mediated by Hypoxia Facilitates Colorectal Cancer Metastasis
    www.nature.com/onc Oncogene ARTICLE OPEN FTO downregulation mediated by hypoxia facilitates colorectal cancer metastasis 1,2,6 1,6 1,3,6 1,6 2 1 1 1 1 1 Dan-Yun Ruan , Ting Li , Ying-Nan Wang , Qi Meng , Yang Li ,KaiYu, Min Wang , Jin-Fei✉ Lin , Li-Zhi Luo , De-Shen✉ Wang , Jun-Zhong Lin4, Long Bai1, Ze-Xian Liu 1, Qi Zhao 1, Xiang-Yuan Wu 2, Huai-Qiang Ju 1,5 and Rui-Hua Xu 1,5 © The Author(s) 2021 Fat mass and obesity-associated protein (FTO), an N6-methyladenosine (m6A) demethylase, participates in tumor progression and metastasis in many malignancies, but its role in colorectal cancer (CRC) is still unclear. Here, we found that FTO protein levels, but not RNA levels, were downregulated in CRC tissues. Reduced FTO protein expression was correlated with a high recurrence rate and poor prognosis in resectable CRC patients. Moreover, we demonstrated that hypoxia restrained FTO protein expression, mainly due to an increase in ubiquitin-mediated protein degradation. The serine/threonine kinase receptor associated protein (STRAP) might served as the E3 ligase and K216 was the major ubiquitination site responsible for hypoxia-induced FTO degradation. FTO inhibited CRC metastasis both in vitro and in vivo. Mechanistically, FTO exerted a tumor suppressive role by inhibiting metastasis-associated protein 1 (MTA1) expression in an m6A-dependent manner. Methylated MTA1 transcripts were recognized by an m6A “reader”, insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2), which then stabilized its mRNA. Together, our findings highlight the critical role of FTO in CRC metastasis and reveal a novel epigenetic mechanism by which the hypoxic tumor microenvironment promotes CRC metastasis.
    [Show full text]
  • A 16Q22.1 Variant Confers Susceptibility to Colorectal Cancer As a Distal Regulator of ZFP90
    Oncogene (2020) 39:1347–1360 https://doi.org/10.1038/s41388-019-1055-4 ARTICLE A 16q22.1 variant confers susceptibility to colorectal cancer as a distal regulator of ZFP90 1 1 2 2 1 1 1 Chen-Yang Yu ● Ji-Xuan Han ● Junfang Zhang ● Penglei Jiang ● Chaoqin Shen ● Fangfang Guo ● Jiayin Tang ● 1 1 1 1 1 1 1,3 4 Tingting Yan ● Xianglong Tian ● Xiaoqiang Zhu ● Dan Ma ● Ye Hu ● Yuanhong Xie ● Wan Du ● Ming Zhong ● 4 5 1,3 1 3 1 1 Jinxian Chen ● Qiang Liu ● Danfeng Sun ● Yingxuan Chen ● Weiping Zou ● Jie Hong ● Haoyan Chen ● Jing-Yuan Fang1 Received: 12 July 2019 / Revised: 1 October 2019 / Accepted: 2 October 2019 / Published online: 22 October 2019 © The Author(s) 2019. This article is published with open access Abstract Genome-wide association studies (GWASs) implicate 16q22.1 locus in risk for colorectal cancer (CRC). However, the underlying oncogenic mechanisms remain unknown. Here, through comprehensive filtration, we prioritized rs7198799, a common SNP in the second intron of the CDH1, as the putative causal variant. In addition, we found an association of CRC- risk allele C of rs7198799 with elevated transcript level of biological plausible candidate gene ZFP90 via expression 1234567890();,: 1234567890();,: quantitative trait loci analysis. Mechanistically, causal variant rs7198799 resides in an enhancer element and remotely regulate ZFP90 expression by targeting the transcription factor NFATC2. Remarkably, CRISPR/Cas9-guided single- nucleotide editing demonstrated the direct effect of rs7198799 on ZFP90 expression and CRC cellular malignant phenotype. Furthermore, ZFP90 affects several oncogenic pathways, including BMP4, and promotes carcinogenesis in patients and in animal models with ZFP90 specific genetic manipulation.
    [Show full text]
  • Predominant Requirement of Bax for Apoptosis in HCT116 Cells Is Determined by Mcl-1’S Inhibitory Effect on Bak
    Oncogene (2012) 31, 3177–3189 & 2012 Macmillan Publishers Limited All rights reserved 0950-9232/12 www.nature.com/onc ORIGINAL ARTICLE Predominant requirement of Bax for apoptosis in HCT116 cells is determined by Mcl-1’s inhibitory effect on Bak C Wang and RJ Youle Biochemistry Section, Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA The intrinsic mitochondrial apoptotic pathway acts apoptosis induced by certain specific stimuli, at least in through two core pro-apoptotic proteins Bax (Bcl2- human cells. For example, Bak or Bax single knock- associated X protein) and Bak (Bcl2-antagonist/killer down renders HeLa cells resistant to Ngo or cisplatin- 1). Although Bax and Bak seem to have redundant roles in induced apoptosis (Kepp et al., 2007). Bax-deficient or apoptosis, accumulating evidence also suggests that they Bak-knockdown human glioblastoma multiforme tumor might not be interchangeable under certain conditions, at cells are also refractory to UV, staurosporine (STS) and least in some human cell lines. Here we report the doxorubicin (Cartron et al., 2003). The BH3-only generation of Bak knockout as well as BaxBak double protein natural born killer (NBK)/Bcl-2-interacting knockout HCT116 human colon carcinoma cells. We killer (Bik) has been shown to act entirely through Bax show that Bak is dispensable for apoptosis induced by a in human cells (Gillissen et al., 2003), but is solely variety of stimuli including ABT-737 but not for dependent on Bak in mouse cells (Shimazu et al., 2007). fluorouracil-induced apoptosis. In addition, Bax deficiency Unlike Bax-deficient MEF cells, BaxÀ/À HCT116 cells only provides partial protection against camptothecin and are extremely resistant to non-steroidal anti-inflamma- cisplatin-induced apoptosis and no protection against tory drugs (NSAIDs) (Zhang et al., 2000) and tumor killing by Puma or ABT-737 plus Noxa overexpression.
    [Show full text]
  • Cyanidin Chloride Induces Apoptosis by Inhibiting NF-Κb Signaling Through Activation of Nrf2 in Colorectal Cancer Cells
    antioxidants Article Cyanidin Chloride Induces Apoptosis by Inhibiting NF-κB Signaling through Activation of Nrf2 in Colorectal Cancer Cells Da-Young Lee, Sun-Mi Yun, Moon-Young Song, Kiwon Jung and Eun-Hee Kim * College of Pharmacy and Institute of Pharmaceutical Sciences, CHA University, Seongnam, 13488, Korea; [email protected] (D.-Y.L.); [email protected] (S.-M.Y.); [email protected] (M.-Y.S.); [email protected] (K.J.) * Correspondence: [email protected]; Tel.: +82-31-881-7179 Received: 19 February 2020; Accepted: 25 March 2020; Published: 27 March 2020 Abstract: Colorectal cancer (CRC) is the third most common cancer worldwide and a leading cause of cancer-related deaths in developed countries. Anthocyanins are a class of flavonoids, widely distributed in food, exhibiting important biological effects. Cyanidin chloride (CyCl) is the common type of anthocyanin with antioxidative and anti-inflammatory potential. The present study aimed to investigate the molecular mechanisms underlying the chemotherapeutic effects of CyCl in colorectal cancer cells. We found that CyCl treatment induced apoptosis as well as a significant inhibition of cellular proliferation and colony formation in three colon cancer HCT116, HT29, and SW620 cells. In addition, CyCl suppressed nuclear factor-kappa B (NF-κB) signaling and induced the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway in tumor necrosis factor-alpha (TNF-α)-stimulated colon cancer cells. Nrf2 and NF-κB are two key transcription factors regulating antioxidative responses and cellular proliferation, respectively. In this study, knockdown of Nrf2 by small interfering RNA (siRNA) transfection inhibited the effect of CyCl on NF-κB signaling and apoptosis, suggesting that there is functional crosstalk between Nrf2 and NF-κB.
    [Show full text]
  • Evidence for a Connection Between the Mismatch Repair System and the G2 Cell Cycle Checkpoint1
    [CANCER RESEARCH 55, 3721-3725. September l, 1W5] Advances in Brief Evidence for a Connection between the Mismatch Repair System and the G2 Cell Cycle Checkpoint1 Mary T. Hawn, Asad Umar, John M. Carethers, Giancarlo Marra, Thomas A. Kunkel, C. Richard Boland,2 and Minoru Koi Division of Gastroenterology, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan 48109-0586 IM. T. H., J. M. €.,G. M., C. R. B., M. K.I; Gastroenterology Section, Veteran's Affairs Medical Center, Ann Arbor, MI 48105-2399 [M. T. H., J. M. C., G. M., C. K. B., M. K.j; and Laboratories of Molecular Carcinogenesis JM. K.j and Molecular Genetics ¡A.U., 7".A. K.j, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709 Abstract instability has been reported to occur in approximately 10-15% of sporadic colonie neoplasias (2, 3), and has also been linked to a defect The human colon tumor cell line HCT116 is deficient in wild-type in mismatch repair in sporadic colon and endometrial tumor cell lines liMIJII. is defective in mismatch repair (MMR), exhibits microsatellite instability, and is tolerant to JV-methylWV'-nitro-JV-nitrosoguanidine (7-9). Previous work with the human colon cancer cell line HCT116, (MNNG). Transferring a normal copy of li.MI.III on chromosome 3 into which has no normal hMLHl gene product, has shown that this line is the cell line restores MMR activity, stabilizes microsatellite loci, and deficient in MMR (8, 9), exhibits microsatellite instability, and is increases the sensitivity of the cell to MNNG.
    [Show full text]
  • Curcumin Potentiates Antitumor Activity of 5-Fluorouracil in a 3D
    Shakibaei et al. BMC Cancer (2015) 15:250 DOI 10.1186/s12885-015-1291-0 RESEARCH ARTICLE Open Access Curcumin potentiates antitumor activity of 5-fluorouracil in a 3D alginate tumor microenvironment of colorectal cancer Mehdi Shakibaei1*, Patricia Kraehe1, Bastian Popper2, Parviz Shayan3,4, Ajay Goel5 and Constanze Buhrmann1 Abstract Background: To overcome the limitations of animal-based experiments, 3D culture models mimicking the tumor microenvironment in vivo are gaining attention. Herein, we investigated an alginate-based 3D scaffold for screening of 5-fluorouracil (5-FU) or/and curcumin on malignancy of colorectal cancer cells (CRC). Methods: The potentiation effects of curcumin on 5-FU against proliferation and metastasis of HCT116 cell and its corresponding isogenic 5-FU-chemoresistant cells (HCT116R) were examined in a 3D-alginate tumor model. Results: CRC cells encapsulated in alginate were able to proliferate in 3D-colonospheres in a vivo-like phenotype and invaded from alginate. During cultivation of cells in alginate, we could isolate 3 stages of cells, (1) alginate proliferating (2) invasive and (3) adherent cells. Tumor-promoting factors (CXCR4, MMP-9, NF-κB) were significantly increased in the proliferating and invasive compared to the adherent cells, however HCT116R cells overexpressed factors in comparison to the parental HCT116, suggesting an increase in malignancy behavior. In alginate, curcumin potentiated 5-FU-induced decreased capacity for proliferation, invasion and increased more sensitivity to 5-FU of HCT116R compared to the HCT116 cells. IC50 for HCT116 to 5-FU was 8nM, but co-treatment with 5 μM curcumin significantly reduced 5-FU concentrations in HCT116 and HCT116R cells (0.8nM, 0.1nM, respectively) and these effects were accompanied by down-regulation of NF-κB activation and NF-κB-regulated gene products.
    [Show full text]