TLR4调控肿瘤增殖相关蛋白CDK4、CDK6对HBV相关性肝癌细胞增殖的作用.pdfVIP

ABSTRACT ABSTRACT Purpose To explore the effects of TLR4 regulating the proliferation of tumor-related protein CDK4,CDK6 on proliferation in HBV-related Hepatic carcinoma. Methods 1.The expression of TLR4,CDK4 and CDK6 protein in 36 cases of HBV-related Hepatic carcinoma tissues and corresponding adjacent tissue were detected by Western blot, and the correlation between them in carcinoma tissues were analyzed. 2.Three specific interference TLR4 siRNA sequence and a negative control sequence were builded and transfected into Hepatoma cells Hep-3B by liposomes.The expression of TLR4 at protein level was measured by Western blot assay before and after transfection, then selecting the best interference sequence. 3.The experiment were divided into the si-RNA(-) group,TLR4-siRNA group and the control group （normal Hep-3B). The expression of TLR4,CDK4 and CDK6 at protein level were measured by Western blot assay before and after transfection.The proliferation in Hepatoma cells was also observed by MTT assay and colony experimental after transfection successfully. Results 1.The overall level of expression of TLR4,CDK4 and CDK6 proteins in HBV-related Hepatic carcinoma were significantly higher than those in the corresponding adjacent tissues(P0.05).And there was a positive correction between TLR4 and CDK4 with correlation coefficient (r) was 0.66 (P0.05), the positive correction between TLR4 and CDK6 was also found (r=0.57,P0.05). 2.Compared with the other two interfere sequences or normal cell, the best effect in reducing the protein levels of TLR4 was obtained with using the TLR4-siRNA-03 interference sequence(P0.05). 3.The proteins levels of CDK