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Open Access Highly Accessed Research article

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

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 and Xiaozhou He1*

Author Affiliations

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

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BMC Cancer 2012, 12:546  doi:10.1186/1471-2407-12-546

Published: 23 November 2012

Abstract

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.

Keywords:
MicroRNA-99a; mTOR; Renal cell carcinoma