miR-29c-3p通过靶向结合PLAU抑制胆管癌细胞增殖、迁移和侵袭

miR-29c-3p inhibits proliferation,migration,and invasion of cholangiocarcinoma cells by targeting PLAU

为探讨miR-29c-3p靶向尿激酶型纤溶酶原激活物(urokinase-type plasminogen activator,PLAU)抑制胆管癌细胞的增殖、迁移和侵袭,构建稳定过表达miR-29c-3p的HuCCT1、 HCCC9810细胞系,将细胞分成miR-NC组、 miR-29c-3p组和miR-29c-3p+PLAU组。生物信息学分析miR-29c-3p、 PLAU的差异表达与胆管癌恶性临床表型之间的关系。采用RT-PCR、 Western blotting检测各组细胞miR-29c-3p、 PLAU的mRNA和蛋白表达水平。CCK-8和Transwell试验检测细胞增殖、迁移和侵袭能力。生物信息学预测miR-29c-3p的靶基因,并用双荧光素酶报告基因试验进行验证。建立胆管癌裸鼠移植瘤模型,分为miR-NC组、 miR-29c-3p组,记录裸鼠肿瘤体积、质量。结果显示,miR-29c-3p在胆管癌中低表达,miR-29c-3p过表达可抑制胆管癌细胞增殖、迁移和侵袭,并可抑制胆管癌裸鼠移植瘤生长(P<0.05)。PLAU在胆管癌中高表达,且PLAU异常表达与恶性临床表型有关。与miR-NC组比较,miR-29c-3p组中miR-29c-3p表达水平升高(P<0.05)。数据库预测PLAU是miR-29c-3p的靶基因,双荧光素酶报告基因试验证实PLAU是miR-29c-3p的作用靶点。miR-29c-3p靶向PLAU抑制胆管癌细胞增殖、迁移和侵袭的能力(P<0.05)。该研究提示,miR-29c-3p可以靶向负调控PLAU的表达,从而抑制胆管癌细胞的增殖、迁移和侵袭。

In order to explore the mechanism of miR-29c-3p on the proliferation,migration,and invasion of cholangiocarcinoma cells by targeting urokinase-type plasminogen activator(PLAU),HuCCT1 and HCCC9810 cell lines with stable overexpression of miR-29c-3p were constructed and divided into miR-NC group,miR-29c-3p group,and miR-29c-3p+PLAU group.The differential expressions of miR-29c-3p and PLAU and their relationships with malignant clinical phenotypes of cholangiocarcinoma were analyzed by bioinformatics.The mRNA and protein expressions of miR-29c-3p and PLAU in each group were detected by RT-PCR and Western blotting,respectively.The proliferation,migration,and invasion of cells were detected by CCK-8 and Transwell assay,respectively.The target gene of miR-29c-3p was predicted by bioinformatics,and verified by dual luciferase reporter gene assay.The xenograft models of cholangiocarcinoma nude mice were constructed and divided into miR-NC group and miR-29c-3p group.The volume and mass of tumors were recorded.The results showed that the expression of miR-29c-3p was down-regulated in cholangiocarcinoma.Overexpression of miR-29c-3p inhibited the proliferation,migration,and invasion of cholangiocarcinoma cells,and the growth of xenografted tumor in nude mice(P<0.05).The expression of PLAU was up-regulated in cholangiocarcinoma,and abnormal expression of PLAU was correlated with malignant clinical phenotypes.Compared to that of the miR-NC group,the expression of miR-29c-3p was increased in miR-29c-3p group(P<0.05).Bioinformatics predicted that PLAU was the target gene of miR-29c-3p,which was verified by dual luciferase reporter gene assay.miR-29c-3p inhibited the proliferation,migration,and invasion of cholangiocarcinoma cells by targeting PLAU(P<0.05).This study suggests that miR-29c-3p can inhibit the proliferation,migration,and invasion of cholangiocarcinoma cells by negatively regulating the expression of PLAU.