MicroRNA-99a促进结肠癌细胞的增殖和迁移及其可能机制

MicroRNA-99a promotes proliferation and migration of colon cancer cell and its anti-tumor mechanism

目的:探讨结肠癌患者组织中microRNA-99a表达水平以及对结肠癌细胞增殖和迁移的影响。方法:取南京医科大学附属常州二院胃肠病中心49例结肠癌患者肿瘤组织、癌旁组织(癌旁5cm)标本以及结肠癌细胞HCT-116、HT-29、SW-480、Caco-2和正常结肠上皮细胞HCoe Pic,采用实时荧光定量PCR检测结肠癌患者肿瘤组织和结肠癌细胞中microRNA-99a表达水平;结肠癌细胞株HT-29转染microRNA-99a抑制剂后,CCK-8法检测microRNA-99a对结肠癌细胞增殖的变化;transwell法观察microRNA-99a对结肠癌细胞迁移的影响;Western blotting检测了HT-29中FGFR-3的表达水平。结果:microRNA-99a表达在结肠癌组织中明显高于癌旁组织(6.27±0.48 vs 1.34±0.54,P<0.05)、在肿瘤细胞中明显高于正常结肠上皮细胞(5.48±0.34,7.67±0.24,5.78±0.22,6.28±0.44 vs 1.45±0.37,P<0.05)。转染microRNA-99a抑制剂后,HT-29细胞的增殖和迁移能力均明显下降(P<0.05);同时,HT-29中FGFR-3显著降低(P<0.05)。结论:microRNA-99a在结肠癌组织中高表达,低表达microRNA-99a可减弱结肠癌细胞的增殖和迁移能力,且可能通过FGFR-3信号通路发挥作用。

Objective： To investigate the expression of microRNA-99 a in the tumor tissues of colon cancer patients and its effect on cancer cell proliferation and migration. Methods： 49 pairs of cancer tissue and adjacent tissue（5cm away from cancer tissue） from colon cancer patients that treated in Gastrointestinal Center of Affiliated Changzhou Second Hospital of Nanjing Medical University, and colon cancer cell lines（HT-29,HCT-116,SW480,Caco-2） as well as normal epithelial Hcoe Pic cell line were selected for our research. Quantitative real-time PCR was used to detect the levels of microRNA-99 a in tumor tissues, adjacent tissues and tumor cells; After transfection of mciro RNA-99 a inhibitor, we used CCK-8 to test the cell proliferation, transwell assay to observe the cell migration,and Western lotting to examine the levels of FGFR3 in HT-29 cells. Results： The expression of microRNA-99 a in the cancer tissues was significantly higher than that in normal para-cancerous tissues（6.27±0.48 vs 1.34±0.54, P〈0.05）, and its expression in tumor cells was significantly higher than that in normal colon epithelial cells（5.48 ±0.34, 7.67±0.24, 5.78±0.22, 6.28±0.44 vs 1.45±0.37, P〈0.05）. After microRNA-99 a inhibitor transfection, cell proliferation and migration of HT-29 cells were significantly decreased（P〈0.05）;, in the meanwhile, the m RNA and protein levels of FGFR3 were significantly decreased in HT-29 cells（P〈0.05）. Conclusion： microRNA-99 a was highly expressed in the tumor tissue of colon cancer patients and colon cancer cells, and low expression of microRNA-99 a may weaken the proliferation and migration ability of cancer cells, which might be accomplished through FGFR3 signaling pathway.