2020/2021
靶向TUG1的mir-29c-3p通過調節CAPN7的表達影響膀胱癌細胞的遷移和侵襲 Mir-29c-3p targeting TUG1 affects migration and invasion of bladder cancer cells by regulating CAPN7 expression
《南方醫科大學學報》*, 2020, 40(9): 1325-1331
| 作者 | 余淦、周輝、許凱、孟麗榮、郎斌 |
|---|---|
| 摘要 |
目的
探討長鏈非編碼RNA TUG1影響膀胱癌細胞遷移和侵襲的機制。
方法
逆轉錄即時定量PCR檢測膀胱癌組織和細胞中TUG1和mir-29c-3p的表達水準。在膀胱癌細胞系T24細胞中利用RNA干擾技術下調TUG1的表達後Transwell檢測細胞的遷移和侵襲能力的變化，並檢測CAPN7的表達水準。在膀胱癌細胞系T24細胞中利用mir-29c-3p類似物過表達mir-29c-3p後在轉錄及轉錄後水準檢測CAPN7的表達變化。功能回復試驗驗證CAPN7及TUG1對膀胱癌細胞遷移和侵襲的影響。
結果
TUG1和mir-29c-3p在膀胱癌細胞和組織中分別表達升高（P=0.01）及減低（P<0.01），同時它們的表達呈現負相關關係（P=0.0109，r2=0.4295）。TUG1表達下調後可以抑制膀胱癌細胞T24的遷移和侵襲能力（P<0.01）。mir-29c-3p過表達後可以下調CAPN7的表達水準，CAPN7的表達水準與TUG1在膀胱癌中呈正相關關係（P=0.0139，r2=0.4081），螢光素酶報告試驗驗證了 mir-29c-3p可同時靶向調節TUG1及CAPN7，功能回復試驗驗證了TUG1可以正向調節CAPN7的表達及其對膀胱癌細胞遷移和侵襲的影響（P<0.01）。
結論
靶向TUG1的mir-29c-3p可通過調節CAPN7的表達影響膀胱癌細胞的遷移和侵襲。
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.
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* 同時列入Scopus。