MicroRNA-99a induces G1-phase cell cycle arrest and suppresses tumorigenicity in renal cell carcinoma

BMC Cancer

Li Cui¹  Hua Zhou²  Hu Zhao³  Yaojun Zhou¹  Renfang Xu¹  Xianlin Xu¹  Lu Zheng⁴  Zhong Xue¹  Wei Xia¹  Bo Zhang¹  Tao Ding¹  Yunjie Cao¹  Zinong Tian¹  Qianqian Shi¹  Xiaozhou He¹ 

[1] Department of Urology, The Third Affiliated Hospital of Soochow University, 185 Juqian Street, Changzhou, 213003, China

[2] Department of Nephrology, The Third Affiliated Hospital of Soochow University, Changzhou, China

[3] Department of Urology, The Affiliated Jiangyin Hospital of Southeast University Medical College, Wuxi, China

[4] Comprehensive Laboratory, The Third Affiliated Hospital of Soochow University, Changzhou, China

关键词: Renal cell carcinoma;    mTOR;    MicroRNA-99a;   

Others  :  1080065
DOI  :  10.1186/1471-2407-12-546

 received in 2012-08-31, accepted in 2012-11-19,  发布年份 2012

【 摘 要 】

Background

A growing body of evidence suggests that microRNAs (miRNAs) play an important role in cancer diagnosis and therapy. MicroRNA-99a (miR-99a), a potential tumor suppressor, is downregulated in several human malignancies. The expression and function of miR-99a, however, have not been investigated in human renal cell carcinoma (RCC) so far. We therefore examined the expression of miR-99a in RCC cell lines and tissues, and assessed the impact of miR-99a on the tumorigenesis of RCC.

Methods

MiR-99a levels in 40 pairs of RCC and matched adjacent non-tumor tissues were assessed by real-time quantitative Reverse Transcription PCR (qRT-PCR). The RCC cell lines 786-O and OS-RC-2 were transfected with miR-99a mimics to restore the expression of miR-99a. The effects of miR-99a were then assessed by cell proliferation, cell cycle, transwell, and colony formation assay. A murine xenograft model of RCC was used to confirm the effect of miR-99a on tumorigenicity in vivo. Potential target genes were identified by western blotting and luciferase reporter assay.

Results

We found that miR-99a was remarkably downregulated in RCC and low expression level of miR-99a was correlated with poor survival of RCC patients. Restoration of miR-99a dramatically suppressed RCC cells growth, clonability, migration and invasion as well as induced G1-phase cell cycle arrest in vitro. Moreover, intratumoral delivery of miR-99a could inhibit tumor growth in murine xenograft models of human RCC. In addition, we also fond that mammalian target of rapamycin (mTOR) was a direct target of miR-99a in RCC cells. Furthermore, siRNA-mediated knockdown of mTOR partially phenocopied the effect of miR-99a overexpression, suggesting that the tumor suppressive role of miR-99a may be mediated primarily through mTOR regulation.

Conclusions

Collectively, these results demonstrate for the first time, to our knowledge, that deregulation of miR-99a is involved in the etiology of RCC partially via direct targeting mTOR pathway, which suggests that miR-99a may offer an attractive new target for diagnostic and therapeutic intervention in RCC.

【 授权许可】

   
2012 Cui et al.; licensee BioMed Central Ltd.

【 图 表 】

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