Synergistic Killing Action of Suicide Gene System Combined with Medicated Serum of Liuwei Dihuang Bolus on Hepatoma Cells / 广州中医药大学学报

ABSTRACT

Objective To investigate the possibility of establish a combined regimen of Chinese medicine and suicide gene therapy,we observed the killing action of medicated serum of Liuwei Dihuang Bolus (LDB)combined with HSV-tk/GCV suicide gene therapy system on rats hepatoma cell line CBRH7919.Methods The HSV-tk/GCV suicide gene therapy system was constructed and the working solution of ganciclovir (GCV)at 39.2 ?mol/L was detected firstly.Then medicated or non-medicated serum was prepared from SD rats treated with or without LDB by gavage.Meanwhile,the effects of medicated and non-medicated serum on cell proliferation were detected.The control groups without serum added were blank control group,GCV group and suicide gene therapy (SGT)group.Blank serum groups,blank serum + GCV groups,blank serum + SGT groups,medicated serum groups,medicated serum+GCV groups and medicated serum+SGT groups all had two serum concentrations of 5% and 7.5%,respectively.Six double holes were set for each group.CBRH7919/ tk+ and CBRH7919/ tk-were mixed together,and the tk+ cell percentage was adjusted to 0%,5% and 10%,respectively.Then the mixture of CBRH7919/ tk+ and CBRH7919/ tk-was implanted into the 96-hole plates at a density of 3?103 cells/hole,cultured with complete medium for 24 hours,and then treated with medicated or non-medicated serum for 12 hours and sequentially with GCV at 39.2?mol/L for another 60 hours.The number of surrival cells was determined by MTT assay.Q value (the ratio of measured and theoretical pharmacological action)was used to analyze the synergism of Chinese medicine and suicide gene therapyadditive action when 0.85≤Q1.15.Results No cytotoxicity was found when blank serum or medicated serum was below the concentration of 20% (volume fraction).When the concentration of serum was at 5% and 7.5%,the killing action of SGT + medicated serum group was stronger and the cell survival rate was higher those that of medicated serum group and SGT + blank serum group (P1.15).Conclusion HSV-tk/GCV suicide gene therapy system combined with LDB has a synergistic killing action on rats hepatoma cells.