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目的研究微小RNA-34 c在子宫内膜癌中的功能。方法以反转录聚合酶连锁反应法(RT-PCR)检测miR-34c在子宫内膜癌组织及正常子宫内膜组织中的表达。将人子宫内膜癌细胞RL95-2分为实验组(miR-34c)及空白组(空白质粒),以噻唑蓝法(MTT)检测细胞活力,用平板克隆形成实验检测细胞克隆形成能力,以流式细胞术检测细胞周期变化,用免疫印迹技术分析细胞中周期蛋白依赖激酶4(CDK4)、CDK6蛋白表达及视网膜母细胞瘤蛋白(Rb)磷酸化水平。结果 miR-34c在子宫内膜癌组织中的表达为0.45±0.05,在正常子宫内膜组织的表达为1.03±0.10,组间比较差异有统计学意义(P<0.01);细胞周期阻滞在G1期,空白组比例为(57.75±5.45)%,实验组为(78.73±8.62)%明显增加(P<0.01),且细胞中CDK4及CDK6表达量明显降低(P<0.01),Rb磷酸化水平也明显降低(P<0.01)。结论 miR-34c在子宫内膜癌组织中低表达,当提高其表达后能显著抑制子宫内膜癌细胞RL95-2的增殖。
Objective To study the function of microRNA-34c in endometrial cancer. Methods The expression of miR-34c in endometrial carcinoma and normal endometrium was detected by reverse transcription-polymerase chain reaction (RT-PCR). The human endometrial carcinoma cell line RL95-2 was divided into experimental group (miR-34c) and blank group (blank plasmid). Cell viability was detected by MTT assay. Cell clonality was measured by plate clone formation assay The changes of cell cycle were detected by flow cytometry. The expressions of CDK4, CDK6 and Rb phosphorylation were analyzed by Western blotting. Results The expression of miR-34c in endometrial carcinoma was 0.45 ± 0.05, and in normal endometrium was 1.03 ± 0.10, the difference was statistically significant (P <0.01). The cell cycle arrest was The expression of CDK4 and CDK6 in G1 phase was significantly lower than that in control group (57.75 ± 5.45)% and 78.73 ± 8.62% (P <0.01) The level was also significantly lower (P <0.01). Conclusion The expression of miR-34c in endometrial carcinoma is low, and its expression can significantly inhibit the proliferation of endometrial carcinoma cell line RL95-2.