miR-574-3p在结直肠癌中的表达和作用机制研究

目的检测并分析结直肠癌组织和细胞系中miR-574-3p的表达情况及其对结直肠癌细胞发生、发展的影响。方法收集武汉市第三医院11对手术切除结直肠癌患者的癌组织和对应癌旁组织,3种结直肠癌细胞系和1种正常结直肠上皮细胞,通过实时荧光定量聚合酶链反应(qRT-PCR)检测临床样本及结直肠癌细胞系中miR-574-3p的表达水平;通过转染miR-574-3p mimic及相应对照mimic NC实现上调结直肠癌细胞(HCT-8、HCT-116)中miR-574-3p的表达水平,采用CCK-8、EdU、克隆形成实验、流式凋亡实验和流式细胞周期实验检测过表达miR-574-3p后对结直肠癌细胞的增殖、凋亡和细胞周期的影响。结果癌旁组织中miR-574-3p的表达差异倍数显著高于对应癌组织(P<0.05),在3种结直肠癌细胞系中miR-574-3p的表达较正常结肠上皮细胞也显著降低(P<0.05)。CCK-8、EdU、克隆形成实验表明,过表达miR-574-3p抑制结直肠癌细胞的增殖能力。流式凋亡实验结果显示,过表达miR-574-3p促进结直肠癌细胞的凋亡能力。流式细胞周期结果显示,过表达miR-574-3p使结直肠癌细胞发生G₀/G₁期阻滞。结论结直肠癌组织和细胞系中的miR-574-3p均低表达,上调结直肠癌细胞(HCT-8、HCT-116)中miR-574-3p可以抑制细胞增殖,促进凋亡并发生G₀/G₁期阻滞。

Expression and effect of miR-574-3p in patients with colorectal cancer

Objective To investigate the expression of miR-574-3p in colorectal cancer tissues and cell lines and its effect on the development of colorectal cancer cells.Methods Expression of miR-574-3p was detected by qRT-PCR in 11 pairs of colorectal cancer tissues and adjacent non-cancer tissues,3 colorectal cancer cell lines and 1 normal colorectal epithelial cell.The abilities of proliferation,apoptosis and cell cycle of HCT-8,HCT-116 cells were determined by CCK-8 assay,Clone formation assay,EdU and flow cytometry assay after overexpression miR-574-3p with mimics.Results The expression of miR-574-3p in the adjacent non-cancer tissues was significantly higher than that in the colorectal cancer tissues(P<0.05),and the expression of miR-574-3p in the three colorectal cancer cell lines was also significantly lower than that in the normal colorectal epithelial cell(P<0.05).CCK-8,EdU and Clone formation assay showed that overexpression of miR-574-3p inhibited the proliferation of colorectal cancer cells.Flow cytometry results showed that overexpression of miR-574-3p promoted the apoptosis of colorectal cancer cells and caused G₀/G₁ phase arrest in colorectal cancer cells.Conclusion The expression of miR-574-3p is reduced in colorectal cancer tissues and 3 colorectal cancer cell lines.After overexpression miR-574-3p in HCT-8 and HCT-116 can inhibit cell proliferation,promote cell apoptosis and stay in the G₀/G₁ phase.