Yu Gan, Zhou Hui, Xu Kai, Meng Lirong, Lang Bin
Department of Urology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Department of Urology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 Sep 30;40(9):1325-1331. doi: 10.12122/j.issn.1673-4254.2020.09.16.
To investigate the mechanism by which long non-coding RNA TUG1 affects bladder cancer cell migration and invasion.
The expressions of TUG1 and miR-29c-3p were examined by quantitative RT-PCR (qRT-PCR) in 10 bladder cancer tissues and 5 bladder cancer cell lines. Trans-well assay was used to detect the changes in migration and invasion abilities of bladder cancer T24 cells after TUG1 knockdown using RNA interference technique, and the alteration in the expression of CAPN7 was also detected. The expression of CAPN7 was examined in T24 cells overexpressing mir-29c-3p by Western blotting, and luciferase reporter assay was performed to confirm the targeting of miR-29c-3p to TUG1 and CAPN7. The effects of CAPN7 overexpression and sh-TUG1 on the migration and invasion of T24 cells were investigated.
The expression of TUG1 was up-regulated and mir-29c-3p was down-regulated significantly in bladder cancer tissue with a negative correlation between their expressions. TUG1 knockdown significantly inhibited the migration and invasion of T24 cells ( < 0.01). Overexpression of mir-29c-3p in T24 cells obviously down-regulated the expression of CAPN7 protein, whose expression was positively correlated with TUG1 expression (=0.4081, =0.0139). The results of luciferase reporter assay confirmed both TUG1 and CAPN7 as the targets of mir-29c-3p. CAPN7 overexpression could partially reverse the tumor suppressing effect of sh-TUG1 in T24 cells.
Mir-29c-3p targeting TUG1 affects the migration and invasion of bladder cancer cells by regulating the expression of CAPN7.
探讨长链非编码RNA TUG1影响膀胱癌细胞迁移和侵袭的机制。
采用定量逆转录聚合酶链反应(qRT-PCR)检测10例膀胱癌组织和5种膀胱癌细胞系中TUG1和miR-29c-3p的表达。运用Trans-well实验,通过RNA干扰技术敲低TUG1后,检测膀胱癌细胞T24迁移和侵袭能力的变化,并检测钙蛋白酶N7(CAPN7)表达的改变。通过蛋白质免疫印迹法检测过表达miR-29c-3p的T24细胞中CAPN7的表达,并进行荧光素酶报告基因实验,以证实miR-29c-3p对TUG1和CAPN7的靶向作用。研究CAPN7过表达和sh-TUG1对T24细胞迁移和侵袭的影响。
膀胱癌组织中TUG1表达上调,miR-29c-3p表达显著下调,二者表达呈负相关。敲低TUG1可显著抑制T24细胞的迁移和侵袭(<0.01)。T24细胞中过表达miR-29c-3p可明显下调CAPN7蛋白表达,CAPN7表达与TUG1表达呈正相关(=0.4081,=0.0139)。荧光素酶报告基因实验结果证实TUG1和CAPN7均为miR-29c-3p的靶点。CAPN7过表达可部分逆转sh-TUG1对T24细胞的抑癌作用。
miR-29c-3p靶向TUG1,通过调节CAPN7的表达影响膀胱癌细胞的迁移和侵袭。