shRNA targeting PLCε inhibits bladder cancer cell growth in vitro and in vivo.

Urology

Department of Urological Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing, People's Republic of China.

Published: August 2011

Objectives: To investigate the role of phospholipase Cε (PLCε) by silencing PLCε with short hairpin RNA (shRNA) in human bladder cancer cells BIU-87 in vitro and in vivo.

Methods: A PLCε shRNA expression vector was transfected into BIU-87 cells, and the expression of PLCε protein was detected by Western blotting. Cell proliferation was determined using the MTT assay, and the cell cycle was detected using flow cytometry. A tumor xenograft experiment was established to evaluate the tumor growth under the condition of PLCε knockdown, and the expression of PLCε, proliferating cell nuclear antigen, and cyclin D1 were detected by Western blotting or immunohistrochemistry.

Results: PLCε shRNA reduced the protein level of PLCε, leading to marked proliferation inhibition and significant cell cycle arrest. Furthermore, PLCε shRNA reduced the tumor xenograft growth implanted with BIU-87 cells. The protein expression of PLCε, proliferating cell nuclear antigen, and cyclin D1 were downregulated in the bladder tumor xenograft.

Conclusions: The knockdown of PLCε by shRNA could inhibit bladder tumor growth and might be an alternative approach for human bladder cancer therapy.

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Source
http://dx.doi.org/10.1016/j.urology.2011.03.014DOI Listing

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