Inhibitory Effect Of Curcumin Combined With SiRNA-hIAP-2 On Human Renal Carcinoma Cell Line GRC-1

Objective: To observe the effect of combination of hIAP-2 gene silencing and curcumin against renal cell carcinoma in vitro,and the possibility to explore the antitumor effect of herbal extracts of curcumin with RNAi collaborative technology,to provide laboratory basis for the comprehensive treatment of tumor.Methods: We selected different concentration of curcumin to interpose renal cancer cell line GRC-1.Then we detected the cell proliferation ability by MTT method,screening out the most suitable conditions for the effect of curcumin.We used curcumin combined with transfection of siRNA-hIAP-2 interposing renal carcinoma cell line GRC-1,detecting the proliferation ability of the combined effect by MTT assay,detecting the expression level of hIAP-2 mRNA by Q-RT-PCR,detecting the expression level of hIAP-2 protein by WB,detecting the level of apoptosis and morphological character of renal carcinoma cell line GRC-1 by flow cytometry,and DAPI,detecting the invasion ability by Transwell,detecting the expression level of NF-kBp65 protein by WB.Results: Curcumin can inhibit the proliferation of GRC-1 cells in different concentrations and time(P<0.05).The renal carcinoma cell line GRC-1 proliferation dropped,When Curcumin combined with siRNA-hIAP-2 interposed renal carcinoma cell line GRC-1(P<0.05),Fluorescence quantitative PCR results showed that the mRNA expression level of hIAP-2 in renal carcinoma cell line GRC-1 dropped(P<0.05),Western blotting showed that the expression level of renal cancer cell line GRC-1 hIAP-2 protein dropped(P<0.05),DAPI assay showed some shrinkage of the GRC-1 cell nucleus,Flow cytometry results showed that the level of apoptosis in renal cancer cell line GRC-1 elevated(P<0.05),Transwell showed that the invasion level of renal carcinoma cell line GRC-1 dropped(P<0.05),Western blotting showed that the expression level of renal cancer cell line GRC-1 NF-kBp65 protein dropped(P<0.05).Conclusion:1.A certain concentration of curcumin can inhibit the proliferation of GRC-1 cells.2.Curcumin combined with siRNA-GRC-1 can significantly inhibit the expression of hIAP-2 in renal carcinoma cell line GRC-1 and depress the expression of NF-kBp65 protein and induce renal carcinoma cell line GRC-1 apoptosis and inhibit renal carcinoma cell line GRC-1 invasion.