miRNA-375 靶向磷脂酰肌醇激酶-3 催化亚单位 α 对宫颈癌细胞增殖和迁移的影响

miRNA-375 regulates proliferation and migration of cervical cancer cells by targeting PIK3CA

摘要:

目的 探讨miRNA-375 靶向磷脂酰肌醇激酶-3 催化亚单位α（PIK3CA）对宫颈癌细胞增殖和 迁移的影响。方法 选取宫颈癌细胞系SiHa，分别转染miRNA-375 模拟基因、miRNA-375 抑制剂、对照模拟 基因及对照抑制剂，采用RT-PCR 法检测miRNA-375 表达，Western blot 检测PIK3CA 蛋白表达，MTT 法检 测细胞增殖活性，划痕实验检测细胞迁移能力。双荧光素酶报告基因实验验证miRNA-375 与PIK3CA 的靶向 关系。结果 转染miRNA-375 模拟基因后miRNA-375 表达上调，PIK3CA 蛋白表达降低（P <0.05）；转染 miRNA-375 抑制剂后miRNA-375 表达降低，PIK3CA 蛋白表达增强（P <0.05）。转染miRNA-375 模拟基 因后SiHa 细胞增殖活性、迁移能力降低（P <0.05）；转染miRNA-375 抑制剂后SiHa 细胞增殖活性、迁移 能力增强（P <0.05）。miRNA 靶基因预测软件检测结果显示，miRNA-375 能作用于PIK3CA 3'-UTR。与转 染对照模拟基因比较，共转染miRNA-375 模拟基因与PIK3CA-WT 可使SiHa 荧光素酶活性降低（P <0.05）。 结论 miRNA-375 可抑制宫颈癌细胞增殖及迁移，其机制可能与靶向调控PIK3CA 有关。

Abstract:

Objective To evaluate the effect of miRNA-375 on regulating proliferation and migration of cervical cancer cells and potential mechanisms. Methods Cervical cancer cells line SiHa was transfected with miRNA-375 mimics, miRNA-375 inhibitors, mimics control, and vehicle control. Expressin of miRNA-375 and PIK3CA were detected by RT-PCR or Western blot. Cell proliferation and migration activity were detected by MTT and scratch test, respectively. Relationship between miRNA-375 and PIK3CA was identified by dual-luciferase report gene experiment. Results After transfected miRNA-375 mimics, expression level of miRNA-375 in SiHa cells was increased while PIK3CA decreased significantly (P < 0.05), all of which were abolished by treatment of miRNA-375 inhibitors (P < 0.05). miRNA-375 mimics transfection induced significant decrease of cellular proliferation and migration ability of SiHa (P < 0.05), which was reversed by miRNA-375 inhibitors (P < 0.05). The miRNA target genes prediction software test results showed that miRNA-375 might bind with PIK3CA 3'UTR, which was confirmed by the fact that co-transfection miRNA-375 mimics and PIK3CA-wt induced decreased of luciferase activity of SiHa (P < 0.05). Conclusions miRNA-375 regulates proliferation and migration of cervical cancer cells by targeting PIK3CA.