MiR-133b regulates bladder cancer cell proliferation and apoptosis by targeting Bcl-w and Akt1

Cancer Cell International

Bin Wu¹  Xia Wang¹  Dong-hui Fu¹  Qiang Liu¹  Shi-jie Li¹  Zhen-qun Xu¹  Ke-feng Wang¹  Xiao-nan Chen¹ 

[1] Department of Urology, Shengjing Hospital of China Medical University, Shenyang 11004, Liaoning, P.R. China

关键词: Apoptosis;    Proliferation;    Bladder cancer;    Akt1;    Bcl-w;    miR-133b;   

Others  :  1121680
DOI  :  10.1186/s12935-014-0070-3

 received in 2013-12-28, accepted in 2014-07-10,  发布年份 2014

【 摘 要 】

Background

MiR-133b is a muscle-specific microRNA; it has a role in the formation of cardiocytes and the expression of myocardium ion channels by regulating target genes. Many human malignant tumors demonstrate a low expression of miR-133b, as noted in colorectal, lung, esophagus and bladder cancers, but the role of miR-133b in bladder cancer is unknown.

Methods

The expression of miR-133b in clinical bladder cancer specimens and adjacent normal tissues was confirmed by stem-loop RT-PCR. We also analyzed the relationship between miR-133b expression and clinicopathological factors of bladder cancer. Bcl-w and Akt1 protein expression in 41 bladder cancer specimens and adjacent normal tissues was detected by Western blot. After transfection of miR-133b mimics or inhibitor into a T24 human bladder cancer cell line, Bcl-w and Akt1 protein and mRNA expression were examined by Western blot and RT-PCR, respectively. The effect of miR-133b on T24 cell proliferation and apoptosis was measured by CCK-8 tests and flow cytometry, respectively.

Results

The expression of miR-133b in bladder cancer tissues from 41 patients was significantly down-regulated (P < 0.01); low expression of miR-133b was strongly associated with high-grade bladder cancer (P < 0.01). Bcl-w and Akt1 proteins were significantly overexpressed in bladder cancer tissues versus adjacent normal tissues (P < 0.01 for both). The expression of Akt1 and Bcl-w proteins and Akt1 mRNA, in T24 cells was significantly down-regulated or up-regulated after transfection of miR-133b mimics or inhibitor, respectively; however, there was no significant difference in Bcl-w mRNA expression. Transfection of HEK-293 T cells with miR-133b significantly suppressed a luciferase-reporter containing the Bcl-w or Akt 1 3′-untranslated regions. MiR-133b mimics significantly inhibited T24 cell proliferation, as well as increased T24 cell apoptosis (P < 0.05 and P < 0.01, respectively) while the miR-133b inhibitor increased and decreased these, respectively (P < 0.05 for both).

Conclusions

MiR-133b may play a very important role in the proliferation and apoptosis of T24 cells by regulating the expression of Bcl-w and Akt1.

【 授权许可】

   
2014 Wu et al.; licensee Springer

【 图 表 】

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