靶向TUG1的mir-29c-3p通过调节CAPN7的表达影响膀胱癌细胞的迁移和侵袭

Mir-29c-3p targeting TUG1 affects migration and invasion of bladder cancer cells by regulating CAPN7 expression

目的探讨长链非编码RNATUG1影响膀胱癌细胞迁移和侵袭的机制。方法逆转录实时定量PCR检测膀胱癌组织和细胞中TUG1和mir-29c-3p的表达水平。在膀胱癌细胞系T24细胞中利用RNA干扰技术下调TUG1的表达后Transwell检测细胞的迁移和侵袭能力的变化,并检测CAPN7的表达水平。在膀胱癌细胞系T24细胞中利用mir-29c-3p类似物过表达mir-29c-3p后在转录及转录后水平检测CAPN7的表达变化。功能回复试验验证CAPN7及TUG1对膀胱癌细胞迁移和侵袭的影响。结果TUG1和mir-29c-3p在膀胱癌细胞和组织中分别表达升高(P=0.01)及减低(P<0.01),同时它们的表达呈现负相关关系(P=0.0109,r^2=0.4295)。TUG1表达下调后可以抑制膀胱癌细胞T24的迁移和侵袭能力(P<0.01)。mir-29c-3p过表达后可以下调CAPN7的表达水平,CAPN7的表达水平与TUG1在膀胱癌中呈正相关关系(P=0.0139,r2=0.4081),荧光素酶报告试验验证了mir-29c-3p可同时靶向调节TUG1及CAPN7,功能回复试验验证了TUG1可以正向调节CAPN7的表达及其对膀胱癌细胞迁移和侵袭的影响(P<0.01)。结论靶向TUG1的mir-29c-3p可通过调节CAPN7的表达影响膀胱癌细胞的迁移和侵袭。

Objective To investigate the mechanism by which long non-coding RNA TUG1 affects bladder cancer cell migration and invasion.Methods 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.Results 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(P<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(r=0.4081,P=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.Conclusion Mir-29c-3p targeting TUG1 affects the migration and invasion of bladder cancer cells by regulating the expression of CAPN7.