OBJECTIVETo construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines.

METHODSLentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by Lipofectamine(TM) 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry.

RESULTSThe recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05).

CONCLUSIONVHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.

Apoptosis ; Carcinoma, Renal Cell ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics

OBJECTIVETo investigate the expression of miR-223 in clear cell renal cell carcinoma (ccRcc) and its clinical implications.

METHODSQuantitative real-time PCR was employed to detect the levels of miR- 223 expression in ccRcc, pair-matched adjacent normal tissues and different renal cancer cell lines. Transwell migration essay and wound healing essay were used to evaluate the invasion and migration of renal cancer 786-O cells transfected with miR-223 mimics. MTT essay was used to measure the cell proliferation, and the cell cycle changes following the transfection were analyzed with flow cytometry.

RESULTSCompared with the normal tissues, the cancer samples showed up-regulated miR-223 expression, which was associated with tumor size. In 786-O cell cultures, transfection with miR-223 mimics significantly enhanced cell migration (P<0.0001) and growth (P=0.006) and induced G1 cell cycle arrest.

CONCLUSIONmiR-223 promotes renal cancer cell migration and proliferation and may serve as a potential therapeutic target for ccRcc.

Carcinoma, Renal Cell ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Flow Cytometry ; G1 Phase Cell Cycle Checkpoints ; Humans ; MicroRNAs ; metabolism ; Real-Time Polymerase Chain Reaction ; Transfection ; Up-Regulation