miRNA-451在宫颈癌中的表达及对宫颈癌Hela细胞增殖与凋亡影响的作用机制研究

Expression of miRNA 451 in cervical cancer and its effect on the proliferation and apoptosis of cervical cancer Hela cells and its mechanism

目的探讨miRNA-451在宫颈癌中的表达及对宫颈癌Hela细胞增殖与凋亡影响的作用机制。方法取60例宫颈癌患者的宫颈癌组织和相应的癌旁正常组织,定量逆转录聚合酶链反应(qRT-PCR)检测miRNA-451的相对表达量,分析其与宫颈癌患者临床特征的关系。将宫颈癌Hela细胞随机分为miRNA-451-mimics组、miRNA-451-mimics-NC组,分别转染miRNA-451模拟物(miRNA-451-mimics)、miRNA-451-mimics阴性对照,对照组未进行转染。采用CCK-8法检测3组宫颈癌Hela细胞的增殖能力,流式细胞仪检测细胞周期和凋亡情况,蛋白质印迹(Westernblot)法检测磷酸化蛋白激酶B(p-AKT)、B细胞淋巴瘤/白血病-2(Bcl-2)蛋白的相对表达量。结果癌旁正常组织中miRNA-451的相对表达量明显高于宫颈癌组织(P﹤0.01)。不同国际妇产科联盟(FIGO)分期、淋巴结转移情况、分化程度宫颈癌患者宫颈癌组织中miRNA-451相对表达量比较,差异均有统计学意义(P﹤0.05)。随着转染时间的增长,宫颈癌Hela细胞中miRNA-451的相对表达量均明显升高(P﹤0.05),细胞存活率逐渐降低,miRNA-451过表达能够将宫颈癌Hela细胞的周期阻滞于G0/G1期并明显促进细胞凋亡。miRNA-451-mimics组宫颈癌Hela细胞p-AKT、Bcl-2蛋白的相对表达量均明显低于对照组(P﹤0.01)。结论miRNA-451在宫颈癌组织中低表达,且与宫颈癌患者FIGO分期、淋巴结转移、分化程度有关,miRNA-451过表达能通过阻断AKT/Bcl-2信号通路来抑制宫颈癌细胞的增殖并诱导其凋亡。

Objective To explore the expression of miRNA-451 in cervical cancer and its effect on the proliferation and apoptosis of cervical cancer Hela cells and its mechanism.Method A total of 60 cervical cancer patients’cervical cancer tissues and the paired normal tissues adjacent to the tumor were taken for the research.The quantitative reverse transcription-polymerase chain reaction(qRT-PCR)assay was used to detect the relative expression of miRNA-451 in the samples,and their relationship with the clinical characteristics of cervical cancer patients was also analyzed.Cervical can-cer Hela cells were randomly divided into miRNA-451-mimics group and miRNA-451-mimics-NC group,and were trans-fected with miRNA-451-mimics and miRNA-451-mimics negative control,respectively,and the control group was not transfected.The CCK-8 method was used to detect the proliferation of the three groups of cervical cancer Hela cells,the flow cytometer was used to detect the cell cycle and apoptosis,and the Western blot method was used to detect the rela-tive expression levels of phosphoinositide protein kinase B(p-AKT)and B cell lymphoma/leukemia-2(Bcl-2)protein.Re-sult The relative expression of miRNA-451 in normal tissues adjacent to cancer was significantly higher than that in cer-vical cancer tissues(P<0.01).The relative expression of miRNA-451 in cervical cancer tissues of patients with different International Federation of Gynecology and Obstetrics(FIGO)stages,lymph node metastasis,and degree of differentia-tion showed statistically significant differences(P<0.05).The relative expression of miRNA-451 in cervical cancer Hela cells increased significantly,while the survival rate gradually decreased with the increase of transfection time(P<0.05).Overexpression of miRNA-451 could block the cell cycle of cervical cancer Hela cells in G0/G1 phase and significantly promote their apoptosis.The relative expression levels of p-AKT and Bcl-2 protein in cervical cancer Hela cells in the miRNA-451-mimics group were significantly lower than those in the control group(P<0.01).Conclusion miRNA-451 is down-regulation in cervical cancer tissues whose expression is related to the FIGO staging,lymph node metastasis,and differentiation grade in cervical cancer patients.Overexpression of miRNA-451 could inhibit the proliferation of cervical cancer cells by blocking the AKT/Bcl-2 signaling pathway and induce apoptosis of cancer cells.