摘要目的 观察微小RNA(miRNA,miR)-371-5p对人结直肠癌LoVo和SW620细胞增殖和凋亡的影响.方法 构建miR-371-5p过表达慢病毒载体和稳定沉默miR-371-5p基因载体[miR-371-5p/短发卡RNA(shRNA)],并转染LoVo和SW620细胞,反转录-聚合酶链反应(RT-PCR)测定miR-371-5p的表达,细胞计数试剂盒(CCK-8)、平板克隆形成实验、软琼脂集落形成实验、流式细胞仪检测LoVo和SW620细胞的增殖和凋亡.结果 LoVo和SW620细胞转染miR-371-5p后miR-371-5p表达(14.25±0.25、10.01±0.19)高于阴性对照组和空白对照组(1.03±0.02、1.04±0.03;1.00±0.00、1.00±0.00;F=21.426、8.957,P=0.000、0.003),LoVo和SW620细胞转染miR-371-5p/shRNA后miR-371-5p表达(0.08±0.00、0.11±0.01)低于阴性对照组和空白对照组(0.97±0.02、1.04±0.03;1.00±0.00、1.00±0.00;F=15.231、8.537,P=0.000、0.005).LoVo和SW620细胞转染后3d和7d,转染miR-371-5p的细胞增殖水平低于阴性对照组和空白对照组(F=5.932、19.324、7.256、35.846,P=0.003、0.000、0.000、0.000);转染miR-371-5p/shRNA的细胞增殖高于阴性对照组和空白对照组(F=37.342、43.254、43.242、105.231,P=0.003、0.000、0.000、0.000).转染miR-371-5p的LoVo和SW620细胞克隆个数低于阴性对照组和空白对照组(F=5.432、4.023,P=0.004、0.022);转染miR-371-5p/shRNA的LoVo和SW620细胞克隆个数高于阴性对照组和空白对照组(F=3.976,4.732,P=0.015、0.011).转染miR-371-5p的LoVo和SW620细胞集落个数低于阴性对照组和空白对照组(F=5.647、4.023,P=0.007、0.015);转染miR-371-5p/shRNA的LoVo和SW620细胞集落个数高于阴性对照组和空白对照组(F=4.231、4.231,P=0.012、0.013).转染miR-371-5p和miR-371-5p/shRNA的LoVo和SW620细胞凋亡率和阴性对照组和空白对照组比较差异均无统计学意义(F=0.883、1.017、1.165、0.954,P=0.742、0.474、0.483、0.621).结论 miR-371-5p抑制人结直肠癌LoVo和SW620细胞的增殖,对细胞凋亡无明显影响.

abstractsObjective To observe the effect of microRNA (miRNA, miR)-371-5p on human colorectal cancer LoVo and SW620 cell proliferation and apoptosis.Methods Constructed miR-371-5p overexpression lentivirus vector and stable gene silencing miR-371-5p vector [miR-371-5p/short hairpin RNA (shRNA)], and were transfected into LoVo and SW620 cells.Reverse transcriptase-polymerase chain reaction (RT-PCR) detected the expression of miR-371-5p, cell counting kit-8 (CCK-8), colony formation assay, soft agar colony formation assay, flow cytometry detected the proliferation and apoptosis of LoVo and SW620 cells.Results The expression of miR-371-5p in LoVo and SW620 cells transfected miR-371-5p (14.25±0.25, 10.01±0.19) were higher than that of negative control group and blank control group (1.03±0.02, 1.04±0.03;1.00±0.00, 1.00±0.00;F=21.426, 8.957, P=0.000, 0.003), The expression of miR-371-5p in LoVo and SW620 cells transfected miR-371-5p/shRNA (0.08±0.00, 0.11±0.01) were lower than that of negative control group and blank control group cells (0.97±0.02, 1.04±0.03;1.00±0.00, 1.00±0.00;F=15.231, 8.537, P=0.000, 0.005).After LoVo and SW620 cells transfected 3 d and 7 d, the proliferation of miR-371-5p transfected cell was lower than that of negative control group and blank control group (F=5.932, 19.324, 7.256, 35.846, P=0.003, 0.000, 0.000, 0.000);the proliferation of transfected miR-371-5p/shRNA cell was higher than that of negative control group and blank control group (F=37.342, 43.254, 43.242, 105.231, P=0.003, 0.000, 0.000, 0.000).The cell clone number of miR-371-5p transfected LoVo and SW620 were lower than that of negative control group and blank control group cells (F=5.432, 4.023, P=0.004, 0.022);the cell clone number of miR-371-5p/shRNA transfected LoVo and SW620 were higherthan that of negative control group and blank control group cells (F=3.976, 4.732, P=0.015, 0.011).The cell colony number of miR-371-5p transfected LoVo and SW620 were lower than that of negative control group and blank control group cells (F=5.647, 4.023, P=0.007, 0.015);the cell colony number of miR-371-5p/shRNA transfected LoVo and SW620 were higher than that of negative control group and blank control group cells (F=4.231, 4.231, P=0.012, 0.013).The difference of cell apoptosis rate between miR-371-5p transfected cells, miR-371-5p/shRNA transfected cells and the negative control group and blank control group were no statistically significant (F=0.883, 1.017, 1.165, 0.954, P=0.742, 0.474, 0.483, 0.621).Conclusion MiR-371-5p can inhibite the proliferation of colorectal cancer LoVo and SW620 cells, has little effect on apoptosis.