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Knockdown of ribonuclease inhibitor expression with siRNA in non-invasive bladder cancer cell line BIU-87 promotes growth and metastasis potentials

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Abstract

Human ribonuclease inhibitor (RI) is a cytoplasmic acidic protein. RI is constructed almost entirely of leucine-rich repeats, which might be involved in some unknown biological functions like other structurally similar proteins besides inhibiting RNase A and angiogenin activities. Our previous experiments demonstrated that up-regulating RI might effectively inhibit some tumor growth and metastasis. However, the down-regulating RI influence on the tumor does not have any report until now, the mechanisms underlying antitumor of RI have not been fully understood. In this study, the efficient RNA interferences of RI were constructed using a plasmid vector and identified with RT-PCR, Western blot and Immunocytochemistry, then were transfected into non-invasive bladder cancer BIU-87 cells. We demonstrated that knockdown RI expression in BIU-87 cells could obviously change the cell morphology, rearrange the microfilaments and extend the lamellipodia, as well as enhance proliferation, increase migration, invasion and matrix metalloprotease level, and also reduce adhesion in vitro. BALB/C nude mice that were injected with the BIU-87 cells transfected RI siRNA showed a significant facilitation of the tumor with heavier tumor weight, higher density of microvessels, lower nm23-H1 and E-Cadherin expressions than those in the control group. Taken together, these experiments suggest that knockdown of RI could promote growth and metastasis potentials of BIU-87 cells. Our present findings reveal the novel mechanism that anti-tumor effect of RI is also involved in suppressing growth and metastasis, besides antiangiogenesis. The results show that RI may be a therapeutic target protein for bladder cancer and may be of biological importance.

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Acknowledgments

This study was supported by grants from National Natural Science Foundation of China (81071719), Chongqing science and technology commission (CSTC, 2006BB5287, CSTC, 2009BB5273).

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Authors and Affiliations

  1. Department of Cell Biology and Genetics, Chongqing Medical University, 1 Yixueyuan Road, Yuzhong District, Chongqing, 400016, People’s Republic of China

    Junxia Chen, Xi Ou-Yang, Juan Gao, Jun Zhu & Xiaoyan He

  2. Molecular Medicine and Cancer Research Center, Chongqing Medical University, 1 Yixueyuan Road, Yuzhong District, Chongqing, 400016, People’s Republic of China

    Junxia Chen, Xi Ou-Yang, Juan Gao, Jun Zhu & Xiaoyan He

  3. Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, 1 Yixueyuan Road, Yuzhong District, Chongqing, 400016, People’s Republic of China

    Jiang Rong

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  1. Junxia Chen
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  2. Xi Ou-Yang
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  3. Juan Gao
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  4. Jun Zhu
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Correspondence to Junxia Chen.

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Chen, J., Ou-Yang, X., Gao, J. et al. Knockdown of ribonuclease inhibitor expression with siRNA in non-invasive bladder cancer cell line BIU-87 promotes growth and metastasis potentials. Mol Cell Biochem 349, 83–95 (2011). https://doi.org/10.1007/s11010-010-0663-7

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  • DOI: https://doi.org/10.1007/s11010-010-0663-7

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