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

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(P<0.05);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(P<0.05);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(P<0.05)。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。

Experimental research of miR-132 inhibits proliferation and induces apoptosis of ovarian cancer via Ezrin

Objective:To explore the biological function of miR-132 in ovarian cancer and the target.Methods: 22 cases ovarian cancer tissue and non-tumor tissue adjacent were collected,the expression of miR-132 in tumor tissue and non-tumor tissue,normal ovarian epithelial cells and ovarian cancer cell were detected by RT-PCR.The normal ovarian epithelial cells which the expression of miR-132 maximum or minimum were chosen,and they were divided into two groups,respectively with transfection of negative control plasmid (NC) and miR-132 mimic plasmid-The expression of miR-132 after transfection was detected by RT-PCR,the cell proliferation and cell apoptosis were detected by CCK-8 method and flow cytometry instrument respectively,the expression of Ezrin protein was detected by Western blot.Results: The expression of miR-132 in tumor tissue was significantly lower than the tumor tissue adjacent,the expression of miR-132 in ovarian cancer cell lines was significantly lower than normal ovarian epithelial cells,the differences were statistically significant (P<0.05).The SKOV3 cell lines was chosed for gene transfection,compared with NC group,transfection with miR-132 mimic plasmid could significantly reduce cell proliferation, increase cell apoptosis, the difference had statistical significance (P<0.05).Western blot results showed that up-regulation miR-132 significantly increased the Ezrin protein expression in ovarian cancer SKOV3 cells (P <0.05).Conclusion: In ovarian cancer,miR-132;inhibits proliferation and induces apoptosis of ovarian cancer via Ezrin,it may be a tumor suppressor gene.