环状RNA circTCF25通过miR-103a-3p/miR-107调控CDK6的表达促进膀胱癌的增殖和迁移

CircRNA circTCF25 Regulates the Expression of CDK6 by miR-103a-3p and miR-107 to Promote the Proliferation and Migration of Bladder Cancer

该文旨在探讨环状RNA circTCF25对膀胱癌增殖和迁移的影响。采用RT-q PCR检测膀胱癌及癌旁组织中miR-103a-3p和miR-107的表达水平;采用Western blot和免疫组化检测膀胱癌细胞及膀胱癌和癌旁组织中CDK6的蛋白表达;将circTCF25过表达载体转染两种膀胱癌细胞后,采用RT-q PCR检测细胞中circTCF25以及miR-103a-3p和miR-107的表达水平;采用双荧光素酶报告基因实验验证circTCF25靶向结合miR-107及miR-107的靶基因CDK6;划痕实验检测细胞迁移能力;Edu实验检测细胞增殖能力。RT-q PCR结果表明,膀胱癌组织中miR-103a-3p和miR-107的表达明显低于癌旁组织。免疫组化和Western blot结果显示,膀胱癌组织中CDK6蛋白质水平明显高于癌旁组织。转染circTCF25过表达载体的细胞中,circTCF25的表达水平高于对照组。过表达circTCF25后,细胞中miR-103a-3p和miR-107的表达显著下降,CDK6的蛋白水平增加。双荧光素酶报告基因实验表明,circTCF25可以直接结合miR-107并降低其对靶基因CDK6的抑制。过表达circTCF25后,细胞迁移和增殖能力增强。该研究说明,环状RNA circTCF25可通过miR-103a-3p/miR-107调控CDK6的表达促进膀胱癌的增殖和迁移。

The aim of the present study is to investigate the effect of circRNA circTCF25 on proliferation and migration of bladder cancer. The expression of miR-103 a-3 p and miR-107 in cancer tissues and adjacent noncancer tissues was determined by RT-q PCR. The expression of CDK6 in bladder cancer cell lines and cancer tissues and their adjacent non-cancer tissues was examined by Western blot and immunohistochemical staining. The circTCF25 over-expression vector was transfected into two kinds of bladder cancer cell lines, and the expression of circTCF25, miR-103 a-3 p and miR-107 were determined by RT-q PCR. Dual luciferase reporter gene assay was used to verify the combination of circTCF25 and miR-107 and the target gene CDK6 of miR-107. The abilities of migration and proliferation of the bladder cancer cell lines were determined by wound healing assay and Edu assay, respectively. As a result, lower expression of miR-103 a-3 p and miR-107, while higher expression of CDK6 was found in bladder cancer tissues compared to their adjacent non-cancer tissues. The expression of circTCF25 in the group transfected with over-expression vector was higher than control group. After up regulation of circTCF25, the expression of miR-103 a-3 p and miR-107 were decreased, while the protein level of CDK6 was increased. Dual luciferase reporter gene assay showed that circTCF25 could directly bind to miR-107 and reduce the inhibition of target gene CDK6. And the capabilities of proliferation and migration of bladder cancer cell lines were increased after over expression of circTCF25. Taken together, circTCF25 may promote the proliferation and migration of bladder cancer through miR-103 a-3 p/miR-107-CDK6 axes.