miR-200c as a Biomarker for 5-FU Chemosensitivity in Colorectal Cancer
5-FU is a chemotherapeutic agent that has been used in colorectal cancer (CRC) treatment. However, it is usually associated with the acquired resistance, which decreases the therapeutic effects of 5-FU. miR-200c is involved in chemotherapeutic drug resistance, but its mechanism is not fully understood. In this study, the effect of inhibition of miR-200c in sensitivity of HCT-116 CRC cells to 5-FU was evaluated. HCT-116 cells were transfected with LNA-anti- miR-200c for 48 h. mRNA expression of miR-200c was evaluated using quantitative real- time PCR. The protein expression of phosphatase and tensin homolog (PTEN) and E-cadherin were analyzed by western blotting. Annexin V and propidium iodide staining assay were applied for apoptosis detection. The caspase-3 activation was evaluated by an enzymatic assay. The results showed LNA-anti-miR-200c inhibited the expression of PTEN and E-cadherin protein, apoptosis and activation of caspase 3 compared with control cells. In conclusion, these results suggest that miR-200c as a prognostic marker can overcome to 5-FU chemoresistance in CRC.
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 Deng J, Lei W, Fu J-C, Zhang L, Li J-H, Xiong J-P. Targeting miR-21 enhances the sensitivity of human colon cancer HT-29 cells to chemoradiotherapy in vitro. Biochemical and biophysical research communications. 2014;443(3):789-95.
 Draht MX, Riedl RR, Niessen H, Carvalho B, Meijer GA, Herman JG et al. Promoter CpG island methylation markers in colorectal cancer: the road ahead. Epigenomics. 2012;4(2):179-94.
 Davoodi H, Hashemi SR, Seow HF. 5-Fluorouracil induce the expression of TLR4 on HCT116 colorectal cancer cell line expressing different variants of TLR4. Iranian journal of pharmaceutical research: IJPR. 2013;12(2):453.
 Boyer J, McLean EG, Aroori S, Wilson P, McCulla A, Carey PD et al. Characterization of p53 wild-type and null isogenic colorectal cancer cell lines resistant to 5-fluorouracil, oxaliplatin, and irinotecan. Clinical Cancer Research. 2004;10(6):2158-67.
 Hsu H-C, Liu Y-S, Tseng K-C, Hsu C-L, Liang Y, Yang T-S et al. Overexpression of Lgr5 correlates with resistance to 5-FU-based chemotherapy in colorectal cancer. International journal of colorectal disease. 2013;28(11):1535-46.
 Wang W, Cassidy J, O’Brien V, Ryan KM, Collie-Duguid E. Mechanistic and predictive profiling of 5-Fluorouracil resistance in human cancer cells. Cancer research. 2004;64(22):8167-76.
 Leichman L, Lenz H-J, Leichman C, Groshen S, Danenberg K, Baranda J et al. Quantitation of intratumoral thymidylate synthase expression predicts for resistance to protracted infusion of 5-fluorouracil and weekly leucovorin in disseminated colorectal cancers: preliminary report from an ongoing trial. European Journal of Cancer. 1995;31(7):1306-10.
 Lau M, Klausen C, Leung P. E-cadherin inhibits tumor cell growth by suppressing PI3K/Akt signaling via β-catenin-Egr1-mediated PTEN expression. Oncogene. 2011;30(24):2753-66.
 Bushati N, Cohen SM. microRNA functions. Annu Rev Cell Dev Biol. 2007;23:175-205.
 Li Z, Wang L, Zhang W, Fu Y, Zhao H, Hu Y et al. Restoring E-cadherin-mediated cell–cell adhesion increases PTEN protein level and stability in human breast carcinoma cells. Biochemical and biophysical research communications. 2007;363(1):165-70.
 Slaby O, Jancovicova J, Lakomy R, Svoboda M, Poprach A, Fabian P et al. Expression of miRNA-106b in conventional renal cell carcinoma is a potential marker for prediction of early metastasis after nephrectomy. Journal of Experimental & Clinical Cancer Research. 2010;29(1):1.
 Philippidou D, Schmitt M, Moser D, Margue C, Nazarov PV, Muller A et al. Signatures of microRNAs and selected microRNA target genes in human melanoma. Cancer research. 2010;70(10):4163-73.
 Karakatsanis A, Papaconstantinou I, Gazouli M, Lyberopoulou A, Polymeneas G, Voros D. Expression of microRNAs, miR‐21, miR‐31, miR‐122, miR‐145, miR‐146a, miR‐200c, miR‐221, miR‐222, and miR‐223 in patients with hepatocellular carcinoma or intrahepatic cholangiocarcinoma and its prognostic significance. Molecular carcinogenesis. 2013;52(4):297-303.
 Cochrane DR, Spoelstra NS, Howe EN, Nordeen SK, Richer JK. MicroRNA-200c mitigates invasiveness and restores sensitivity to microtubule-targeting chemotherapeutic agents. Molecular cancer therapeutics. 2009;8(5):1055-66.
 Ceppi P, Mudduluru G, Kumarswamy R, Rapa I, Scagliotti GV, Papotti M et al. Loss of miR-200c expression induces an aggressive, invasive, and chemoresistant phenotype in non–small cell lung cancer. Molecular Cancer Research. 2010;8(9):1207-16.
 Feng X, Wang Z, Fillmore R, Xi Y. MiR-200, a new star miRNA in human cancer. Cancer letters. 2014;344(2):166-73.
 Chen J, Wang W, Zhang Y, Hu T, Chen Y. The roles of miR-200c in colon cancer and associated molecular mechanisms. Tumor Biology. 2014;35(7):6475-83.
 Sharifi M, Salehi R, Gheisari Y, Kazemi M. Inhibition of microRNA miR-92a induces apoptosis and inhibits cell proliferation in human acute promyelocytic leukemia through modulation of p63 expression. Molecular biology reports. 2014;41(5):2799-808.
 Ghaznavi H, Najafi R, Mehrzadi S, Hosseini A, Tekyemaroof N, Shakeri-zadeh A et al. Neuro-protective effects of cerium and yttrium oxide nanoparticles on high glucose-induced oxidative stress and apoptosis in undifferentiated PC12 cells. Neurological research. 2015;37(7):624-32.
 Wiklund ED, Bramsen JB, Hulf T, Dyrskjøt L, Ramanathan R, Hansen TB et al. Coordinated epigenetic repression of the miR‐200 family and miR‐205 in invasive bladder cancer. International journal of cancer. 2011;128(6):1327-34.
 Davalos V, Moutinho C, Villanueva A, Boque R, Silva P, Carneiro F et al. Dynamic epigenetic regulation of the microRNA-200 family mediates epithelial and mesenchymal transitions in human tumorigenesis. Oncogene. 2012;31(16):2062-74.
 Magee P, Shi L, Garofalo M. Role of microRNAs in chemoresistance. Annals of translational medicine. 2015;3(21).
 Chen Y, Sun Y, Chen L, Xu X, Zhang X, Wang B et al. miRNA-200c increases the sensitivity of breast cancer cells to doxorubicin through the suppression of E-cadherin-mediated PTEN/Akt signaling. Molecular medicine reports. 2013;7(5):1579-84.
 Yin Y, Shen W. PTEN: a new guardian of the genome. Oncogene. 2008;27(41):5443-53.
 Lu Y-X, Yuan L, Xue X-L, Zhou M, Liu Y, Zhang C et al. Regulation of Colorectal Carcinoma Stemness, Growth, and Metastasis by an miR-200c-Sox2–Negative Feedback Loop Mechanism. Clinical Cancer Research. 2014;20(10):2631-42.
 Liao C, Chen W, Fan X, Jiang X, Qiu L, Chen C et al. MicroRNA-200c inhibits apoptosis in pituitary adenoma cells by targeting the PTEN/Akt signaling pathway. Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics. 2014;21(3):129-36.
 Berlanga P, Muñoz L, Piqueras M, Sirerol JA, Sánchez-Izquierdo MD, Hervás D et al. miR-200c and phospho-AKT as prognostic factors and mediators of osteosarcoma progression and lung metastasis. Molecular oncology. 2016.
 Vivanco I, Sawyers CL. The phosphatidylinositol 3-kinase–AKT pathway in human cancer. Nature Reviews Cancer. 2002;2(7):489-501.
 Oki E, Baba H, Tokunaga E, Nakamura T, Ueda N, Futatsugi M et al. Akt phosphorylation associates with LOH of PTEN and leads to chemoresistance for gastric cancer. International journal of cancer. 2005;117(3):376-80.
 Hur K, Toiyama Y, Takahashi M, Balaguer F, Nagasaka T, Koike J et al. MicroRNA-200c modulates epithelial-to-mesenchymal transition (EMT) in human colorectal cancer metastasis. Gut. 2013;62(9):1315-26.
 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. methods. 2001;25(4):402-8.
 Boominathan L. The tumor suppressors p53, p63, and p73 are regulators of microRNA processing complex. PloS one. 2010;5(5):e10615.
 Sun J, Ding W, Zhi J, Chen W. MiR-200 suppresses metastases of colorectal cancer through ZEB1. Tumor Biology. 2015:1-7.
 Karimi Dermani F, Saidijam M, Amini R, Mahdavinezhad A, Heydari K, Najafi R. Resveratrol Inhibits Proliferation, Invasion, and Epithelial–Mesenchymal Transition by Increasing miR‐200c Expression in HCT‐116 Colorectal Cancer Cells. Journal of Cellular Biochemistry. 2016.
 Kim SM, Kim JS, Kim J-H, Yun C-O, Kim EM, Kim HK et al. Acquired resistance to cetuximab is mediated by increased PTEN instability and leads cross-resistance to gefitinib in HCC827 NSCLC cells. Cancer letters. 2010;296(2):150-9.
 Takamizawa J, Konishi H, Yanagisawa K, Tomida S, Osada H, Endoh H et al. Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival. Cancer research. 2004;64(11):3753-6.
 Gotanda K, Hirota T, Matsumoto N, Ieiri I. MicroRNA-433 negatively regulates the expression of thymidylate synthase (TYMS) responsible for 5-fluorouracil sensitivity in HeLa cells. BMC cancer. 2013;13(1):369.
 Hewish M, Lord CJ, Martin SA, Cunningham D, Ashworth A. Mismatch repair deficient colorectal cancer in the era of personalized treatment. Nature reviews Clinical oncology. 2010;7(4):197-208.
 Bronckaers A, Gago F, Balzarini J, Liekens S. The dual role of thymidine phosphorylase in cancer development and chemotherapy. Medicinal research reviews. 2009;29(6):903-53.
 Holzner S, Senfter D, Stadler S, Staribacher A, Nguyen CH, Gaggl A et al. Colorectal cancer cell-derived microRNA200 modulates the resistance of adjacent blood endothelial barriers in vitro. Oncology Reports. 2016;36(5):3065-71.
 Chen J, Wang W, Zhang Y, Chen Y, Hu T. Predicting distant metastasis and chemoresistance using plasma miRNAs. Medical oncology. 2014;31(1):1-7.
 Toden S, Okugawa Y, Jascur T, Wodarz D, Komarova NL, Buhrmann C et al. Curcumin mediates chemosensitization to 5-fluorouracil through miRNA-induced suppression of epithelial-to-mesenchymal transition in chemoresistant colorectal cancer. Carcinogenesis. 2015;36(3):355-67.
 Bonci D, Coppola V, Musumeci M, Addario A, Giuffrida R, Memeo L et al. The miR-15a–miR-16-1 cluster controls prostate cancer by targeting multiple oncogenic activities. Nature medicine. 2008;14(11):1271-7.