MicroRNA-143 inhibits proliferation and migration as well as invasion in esophageal cancer cell line ECA109

doi:10.3760/cma.j.issn.1673-422X.2016.11.001

Abstract

Abstract: Objective To investigate the functions of microRNA143 (miR143) in esophageal cancer cell line ECA109. MethodsECA109 cells were transfected with negative control (NC), miR143 mimics or miR143 inhibitors. 3(4,5dimethyl2thiazolyl)2,5diphenyl2Htetrazolium bromide (MTT) assay was performed to evaluate the growth of ECA109 cells after transfection. Annexin VFITC/PI apoptosis test kit was used to detect early apoptosis rate in ECA109 cells. Transwell migration and invasion assays were conducted to compare the migration and invasion capacity of ECA109 among different groups. Realtime PCR and Western blotting were used to analyze the mRNA and protein alteration after transfection. ResultsThree and four days after transfection, compared with NC (absorbance value: 0.90±0.02 and 1.09±0.07), miR143 mimics inhibited ECA109 cell proliferation (absorbance value: 0.66±0.05 and 0.80±0.04), while miR143 inhibitors promoted cell proliferation (absorbance value: 1.13±0.09 and 1.51±0.08), with statistical significances (F=49.16, P=0.000; F=100.34, P=0.000). Earlystage apoptosis rates of ECA109 transfected with NC, miR143 mimics and miR143 inhibitors were 3.42%±0.72%, 11.63%±1.15% and 0.94%±0.10%, respectively, with statistical significance (F=151.61, P=0.000). Meanwhile, compared with NC (migration cell number: 336±13, invasion cell number: 147±16), miR143 mimics inhibited cell migration (148±16) and invasion (75±10), while miR143 inhibitors promoted cell migration (510±14) and invasion (238±16), with statistical significances (F=470.99, P=0.000; F=90.04, P=0.000). Compared with NC (1.00±0.00), miR143 mimics downregulated mRNA (relative expression level 0.22±0.08) and protein expression (relative expression level 0.46±0.08) of Kras, whereas miR143 inhibitors upregulated mRNA (1.55±0.12) and protein expression (1.33±0.05) of Kras (F=131.36, P=0.000; F=88.17, P=0.000). ConclusionmiR143 functions as a tumor suppressor in esophageal cancer cell line ECA109, probably by downregulating Kras expression.

Key words: MicroRNAs, Esophageal neoplasms, Genes, tumor suppressor

Wu Wenxue, Liu Wanping, Liu Hebo, Chen Jie, Sun Yanyan, Zou Chuantao, Li Yanmei. MicroRNA-143 inhibits proliferation and migration as well as invasion in esophageal cancer cell line ECA109[J]. Journal of International Oncology, 2016, 43(11): 801-805.

References

［1］ Mao YS, He J, Cheng GY. Current status of surgical management of esophageal cancer in China and the future strategy［J］. Zhonghua Zhong Liu Za Zhi, 2010, 32(6): 401-404. ［2］ Ha M, Kim VN. Regulation of microRNA biogenesis［J］. Nat Rev Mol Cell Biol, 2014, 15(8): 509-524. DOI: 10.1038/nrm3838. ［3］ Ryan BM, Robles AI, Harris CC. Genetic variation in microRNA networks: the implications for cancer research［J］. Nat Rev Cancer, 2010, 10(6): 389-402. DOI: 10.1038/nrc2867. ［4］ Kohlhapp FJ, Mitra AK, Lengyel E, et al. MicroRNAs as mediators and communicators between cancer cells and the tumor microenvironment［J］. Oncogene, 2015, 34(48): 5857-5868. DOI: 10.1038/onc.2015.89. ［5］ Hong L, Han Y, Zhang H, et al. Prognosisrelated microRNAs in esophageal cancer［J］. Expert Opin Biol Ther, 2014, 14(4): 483-489. DOI: 10.1517/14712598.2014.882896. ［6］ Huang J, Zhang SY, Gao YM, et al. MicroRNAs as oncogenes or tumour suppressors in oesophageal cancer: potential biomarkers and therapeutic targets［J］. Cell Prolif, 2014, 47(4): 277-286. DOI: 10.1111/cpr.12109. ［7］ Wu BL, Xu LY, Du ZP, et al. MiRNA profile in esophageal squamous cell carcinoma: downregulation of miR-143 and miR-145［J］. World J Gastroenterol, 2011, 17(1): 79-88. DOI: 10.3748/wjg.v17.i1.79. ［8］ Liu R, Liao J, Yang M, et al. The cluster of miR143 and miR145 affects the risk for esophageal squamous cell carcinoma through co-regulating fascin homolog 1［J］. PLoS One, 2012, 7(3): e33987. DOI: 10.1371/journal.pone.0033987. ［9］ Ni Y, Meng L, Wang L, et al. MicroRNA-143 functions as a tumor suppressor in human esophageal squamous cell carcinoma［J］. Gene, 2013, 517(2): 197-204. DOI: 10.1016/j.gene.2012.12.031. ［10］ Ansari MH, Irani S, Edalat H, et al. Deregulation of miR-93 and miR-143 in human esophageal cancer［J］. Tumour Biol, 2016, 37(3): 3097-3103. DOI: 10.1007/s13277-015-3987-9. ［11］ Clapé C, Fritz V, Henriquet C, et al. miR143 interferes with ERK5 signaling, and abrogates prostate cancer progression in mice［J］. PLoS One, 2009, 4(10): e7542. DOI: 10.1371/journal.pone.0007542. ［12］ Xu B, Niu X, Zhang X, et al. miR-143 decreases prostate cancer cells proliferation and migration and enhances their sensitivity to docetaxel through suppression of K-RAS［J］. Mol Cell Biochem, 2011, 350(1/2): 207-213. DOI: 10.1007/s11010-010-0700-6. ［13］ Zhou P, Chen WG, Li XW. MicroRNA-143 acts as a tumor suppressor by targeting hexokinase 2 in human prostate cancer［J］. Am J Cancer Res, 2015, 5(6): 2056-2063. ［14］ Xu YF, Li YQ, Guo R, et al. Identification of miR-143 as a tumour suppressor in nasopharyngeal carcinoma based on microRNA expression profiling［J］. Int J Biochem Cell Biol, 2015, 61: 120-128. DOI: 10.1016/j.biocel.2015.02.006. ［15］ Zhai L, Ma C, Li W, et al. miR-143 suppresses epithelialmesenchymal transition and inhibits tumor growth of breast cancer through downregulation of ERK5［J］. Mol Carcinog, 2015: 22445. DOI: 10.1002/mc.22445. ［16］ Wu XL, Cheng B, Li PY, et al. MicroRNA143 suppresses gastric cancer cell growth and induces apoptosis by targeting COX-2［J］. World J Gastroenterol, 2013, 19(43): 7758-7765. DOI: 10.3748/wjg.v19.i43.7758. ［17］ Zhuang M, Shi Q, Zhang X, et al. Involvement of miR-143 in cisplatin resistance of gastric cancer cells via targeting IGF1R and BCL2［J］. Tumour Biol, 2015, 36(4): 2737-2745. DOI: 10.1007/s13277-014-2898-5. ［18］ Chen X, Guo X, Zhang H, et al. Role of miR-143 targeting K-RAS in colorectal tumorigenesis［J］. Oncogene, 2009, 28(10): 1385-1392. DOI: 10.1038/onc.2008.474. ［19］ Su J, Liang H, Yao W, et al. MiR-143 and miR-145 regulate IGF1R to suppress cell proliferation in colorectal cancer［J］. PLoS One, 2014, 9(12): e114420. DOI: 10.1371/journal.pone.0114420. ［20］ Osaki M, Takeshita F, Sugimoto Y, et al. MicroRNA-143 regulates human osteosarcoma metastasis by regulating matrix metalloprotease-13 expression［J］. Mol Ther, 2011, 19(6): 11231130. DOI: 10.1038/mt.2011.53. ［21］ Song T, Zhang X, Wang C, et al. Expression of miR-143 reduces growth and migration of human bladder carcinoma cells by targeting cyclooxygenase2［J］. Asian Pac J Cancer Prev, 2011, 12(4): 929933. ［22］ Von Karstedt S, Conti A, Nobis M, et al. Cancer cellautonomous TRAILR signaling promotes KRASdriven cancer progression, invasion, and metastasis［J］. Cancer Cell, 2015, 27(4): 561-573. DOI: 10.1016/j.ccell.2015.02.014. ［23］ Radojicic J, Zaravinos A, Spandidos DA. HPV, KRAS mutations, alcohol consumption and tobacco smoking effects on esophageal squamouscell carcinoma carcinogenesis［J］. Int J Biol Markers, 2012, 27(1): 1-12. DOI: 10.5301/JBM.2011.8737.

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