Investigate the mechanism of Liuwei Dihuang Bolus enhancing hepatocarcinoma suicide gene therapy from Cx26.

Objective:To investigate the regulation effect of medicated serum of Liuwei Dihuang Bolus on the expression of Connexin26 in rat hepatocarcinoma CBRH7919 cell,and clarity whether the mechanism of Liuwei Dihuang Bolus promoting the suicide gene therapy was related with the gap junction.Methods:Blood-serum pharmacology was used to prepare the medicated serum of Liuwei Dihuang Bolus.The lethal e ect of the medicated serum of Liuwei Dihuang Bolus associated with the HSV-tk/GCV suicide gene therap system was detected by MTT assay.The expression of Cx26 and mRNA on rats hepatoma cell line CBRH7919 were detected by western blot,indirect immuno uorescence assay(IIA)and real-time uorescence quantitative RT-PCR assay.Results:①The survival rate of hepatoma carcinoma cell in medicated serum combining with tk+/ GCV group was signi cantly lower than that in suicide gene therapy group(10%tk+/GCV with control serum)(P0.01),the lethal e ect had obvious cooperativity(Q1.15);②The result of western blot indicated that the medicated serum of Liuwei Dihuang Bolus can elevate the protein expression of Cx26,also the up-regulated e ects showed obvious dose-e ect relationship.③The IIA assay showed similar results:Cx26 protein was located on the cell membrane and cytoplasm,especially more located on cell membrane,and the uorescence intensity of medicated serum group is much stronger than that of control-serum group(P0.05).Conclusion:The mechanism of Liuwei Dihuang Bolus enhancing suicide gene bystander e ect relates to that it can promote the expression Cx26 mRNA and protein on CBRH7919,make more Cx26 located on cell membrane,improve the gap junctional intercellular communication(GJIC).