| [Objective]Renal cell carcinoma (RCC) is the third most common genitourinary malignancy, The mortality and morbidity of RCC is strongly associated with its high propensity to metastasize to specific organs. RNA interference (RNAi), which has been proven to be a powerful tool for suppressing gene expression, may provide a promising way forward in RCC therapy. The recent researches indicated that chemokine recepror CXCR4 plays an important role in the invasion and metastasis of tumor.In the present study, we employed the RNAi technique to inhibit CXCR4 expression in RCC line A498 and investigated the effect of CXCR4 suppression on the proliferation and apoptosis of those cells in vitro.[Methods]We designed and synthesized CXCR4 specific short hairpin RNA (shRNA),The hairpin loop region was annealed with its complementary strand, and was cloned into the pGPU6/GFP/Neo plasmid vector which transfected into renal carcinoma cell line A498 by using Lipofectamine2000.We observed the change of expression of CXCR4 by RT-PCR and western blot analysis and detected the change of metastatic potention by MTT and transwell vitro invasion assay.Analysis of apoptosis and cell cycle were by flow cytometry. All measurements were carried out with the same instrument under the same experimental condition. Data were expressed as means±standard deviation. All data were analyzed statistically by one-way analysis of variance(ANOVA), with P |
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