miR-132靶向调控Ezrin抑制卵巢癌细胞增殖、促进凋亡的实验研究

发布时间：2018-02-01 11:37

本文关键词： miR- 卵巢癌增殖凋亡埃兹蛋白　出处：《中国免疫学杂志》2017年01期 　论文类型：期刊论文

【摘要】：目的:探讨微小核糖核酸分子(miRNA)-132在卵巢癌中生物学作用和作用靶点。方法:收集22例卵巢癌及癌旁非肿瘤组织标本,RT-PCR检测miR-132表达量;RT-PCR检测人正常卵巢上皮细胞和卵巢癌细胞系中miR-132表达量;选择miR-132表达量最高或最低的卵巢癌细胞株,分别转染阴性对照质粒(NC)和miR-132 mimic质粒,RT-PCR检测转染后miR-132表达量;CCK-8法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测Ezrin蛋白表达。结果:卵巢癌组织中miR-132表达量显著低于癌旁非肿瘤组织,而卵巢癌细胞系中miR-132表达量显著低于正常卵巢上皮细胞,差异均有统计学意义(P0.05);选择卵巢癌细胞株中miR-132表达量最低卵巢癌SKOV3细胞株进行基因转染,与转染阴性对照质粒相比,转染miR-132 mimic质粒后miR-132表达量显著上升,细胞增殖显著降低,细胞凋亡显著增加,差异均有统计学意义(P0.05);Western blot结果显示,上调miR-132表达后,卵巢癌SKOV3细胞株中Ezrin蛋白表达显著上升(P0.05)。结论:在卵巢癌中,miR-132可能作为一种抑癌基因通过靶向调控Ezrin抑制卵巢癌细胞增殖、促进凋亡。
[Abstract]:Objective: to investigate the biological role and target of microRNA molecule miRNA-132 in ovarian cancer. Methods: 22 specimens of ovarian cancer and non-cancerous tissues were collected. The expression of miR-132 was detected by RT-PCR. RT-PCR was used to detect the expression of miR-132 in human ovarian epithelial cells and ovarian cancer cell lines. The ovarian cancer cell lines with the highest or lowest miR-132 expression were selected and transfected into negative control plasmids (NCCs) and miR-132 mimic plasmids, respectively. The expression of miR-132 after transfection was detected by RT-PCR. CCK-8 assay was used to detect cell proliferation and flow cytometry was used to detect cell apoptosis. Western blot was used to detect the expression of Ezrin protein. Results: the expression of miR-132 in ovarian cancer tissues was significantly lower than that in non-cancerous adjacent tissues. The expression of miR-132 in ovarian cancer cell line was significantly lower than that in normal ovarian epithelial cell line (P 0.05). The lowest miR-132 expression level of ovarian cancer SKOV3 cell line was selected for gene transfection, compared with the negative control plasmid. After transfection of miR-132 mimic plasmid, the expression of miR-132 increased significantly, cell proliferation decreased and apoptosis increased significantly (P 0.05). The results of Western blot showed that the expression of miR-132 was up-regulated. The expression of Ezrin protein in ovarian cancer SKOV3 cell line increased significantly (P 0.05). Conclusion: in ovarian cancer cell line, the expression of Ezrin protein is significantly increased. As a tumor suppressor gene, miR-132 may inhibit the proliferation and promote apoptosis of ovarian cancer cells by targeting Ezrin.
【作者单位】：蚌埠医学院第一附属医院妇瘤科;
【分类号】：R737.31
【正文快照】： 卵巢癌在妇科肿瘤中发病率排第三,病死率第一。卵巢癌病死率居高不下的原因是由于早期筛查和诊断困难,患者就诊时大部分已经处于中晚期,加上卵巢癌对化疗的耐药以及复发和转移[1]。而提高卵巢癌的早期诊断和有效治疗方法是目前临床亟待解决的问题。埃兹蛋白(Ezrin)是ERM家族成

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