MiR-622抑制人结直肠腺癌细胞HT-29增殖的研究

Research of MiR-622 in inhibiting the proliferation of HT-29 in human colorectal cancer

目的探讨MiR-622对人结直肠腺癌细胞HT-29增殖的影响及其可能的作用机制。方法将培养的人结直肠腺癌HT-29细胞分为三组,空白组、瞬时转染MiR-622模拟物组、瞬时转染MiR-622模拟物阴性对照组,瞬时转染后0、12、24、36、48 h,Q-PCR法分别检测各组细胞MiR-622表达水平,四甲基偶氮唑蓝(MTT)比色法分别检测各组细胞的增殖程度,瞬时转染后24 h,蛋白免疫印迹检测各组细胞k-Ras、p-Mek、p-Erk蛋白的表达水平。结果与空白组比较,模拟物阴性对照组MiR-622表达水平无明显变化(P>0.05),模拟物组Mi R-622表达水平在12、24、36、48 h均明显升高(P<0.01);模拟物阴性对照组细胞增殖程度无明显变化(P>0.05),模拟物组细胞增殖程度受到抑制(P<0.01);模拟物阴性对照组细胞中k-Ras、p-Mek、p-Erk蛋白表达水平无明显变化(P>0.05),模拟物组细胞中k-Ras、p-Mek、p-Erk蛋白表达水平均明显降低(P<0.05)。结论 MiR-622能够降低Ras信号通路的转导,抑制人结直肠腺癌细胞HT-29的增殖。

Objective To research the influence of MiR-622 on the proliferation of HT-29 in human colorectal cancer and explore the possible mechanism. Methods The cultured HT-29 cells were divided into 3 groups：the control group,transient transfection of MiR-622 mimic group and transient transfection of MiR-622 mimic negative control group. The expression levels of MiR-622 were detected by quantitative polymerase chain reaction method on 0,12,24,36,48 h after transient transfection, the proliferation levels of cells in all the groups were detected by MTF method,and the protein expression levels of k-Ras ,p-Mek and p-Erk were detected by Western blotting. Results Compared to the control group,the MiR-622 expression level,proliferation level and protein expression levels of k-Ras,p-Mek and p-Erk in the transient transfection of MiR-622 mimic negative control group had no statistically significant difference （P〉0.05）, but the MiR-622 levels in the MiR-622 mimic group on 12,24,36,48 h increased obviously, the proliferation level was inhibited remarkably, and the protein expression levels of k-Ras, p-Mek and p-Erk reduced apparently, the difference had statistical significance （P〈0.01 or 0.05）. Conclusion MiR-622 can decrease the activity of Ras signaling pathway and inhibit the proliferation of HT-29 in human colorectal cancer.