The Synergistic Effect Of Resveratrol On The Tumour Suicide Gene Therapy

ObjectivesThis research will have to determine the active ingredient in traditional Chinese medicine for suicide gene system with resveratrol synergy study. Explore how its enhanced suicide gene through GJIC bystander effect, for the establishment of Chinese medicine treatment of cancer, anti-cancer medicine to promote clinical application of new drugs and to lay the foundation for cancer gene therapy.MethodsIn this experiment, cultured rat hepatoma cells CBRH7919, CBRH7919TK, mouse malignant melanoma B16-RFP, B16/TK-GFP study. Rat hepatoma suicide gene system using MTT assay synergism of resveratrol on them to determine the effect of time and concentration; Western Blot assay resveratrol connections protein Cx26, Cx32and Cx43expression of the gap; Nude mice with resveratrol experiments on rat liver cancer suicide gene system synergies. Malignant melanoma in mice suicide gene (with green fluorescence) using MTT assay synergism of resveratrol on them to determine the effect of time and concentration. Resveratrol was detected by flow cytometry effect on apoptosis. Western Blot assay resveratrol connections protein Cx26, Cx32and Cx43expression of the gap.Results1. Resveratrol on mouse malignant melanoma synergism suicide gene system.(1) Synergistic effect of resveratrol on30%B16TK-GFP/GCVWith a concentration of4μmol/L、8μmol/L、16μmol/L、32μmol/L resveratrol of30%B16/TK-GFP cells after72h, the inhibition rate was-3.43±3.03%,-1.88±1.29%,26.65±1.87%.36.95±3.23%; A concentration of4μmol/L、8μmol/L、16μmol/L、32μmol/L resveratrol combine with30%B16tk-GFP/GCV cells after72h, the inhibition rate was43.32±3.93%. 47.00±4.13%、53.74±4.61%,55.76±3.83%, Jin Shi Q values was1.98,2.04,1.21、1.06. In addition to32μmol/L combined with30%B16TK-GFP/GCV are additive, other concentrations are synergies.(2) Apoptosis by flow cytometry of resveratrol on30%B16/TK-GFPResveratrol combined30%B16tk, sub ploidy peak of apoptotic cells compare with control from3.00%to4.15%,5.26%,6.96%, When the resveratrol is16μmol/L, compare to the control, there were significant differences (P<0.05), that the pro-apoptotic effects of resveratrol in a concentration range of4μmol/L、8μmol/L、16μmol/L this is not obvious. Resveratrol combined with30%B16tk/GCV, the subGl from6.63%to8.96%,11.32%,15.24%, compared to the GCV, the resveratrol in a concentration range of8μmol/L、16μmol/L, there were significant differences (P<0.01). Jin Shi Q values are more than1.15. that can explain resveratrol when combined GCV suicide gene therapy system, there is a certain synergy.(3) Western Blot detection impact of resveratrol on Cx26, Cx32, Cx43protein expression of B16-RFP cells.B16-RFP cells use resveratrol48h, Cx26protein tended to increase; Cx32protein have no statistical difference; Cx43protein tended to increase.2. Resveratrol on rat hepatoma synergies suicide gene system(1) Synergistic effect of resveratrol on20%CBRH7919TK/GCVWith a concentration of2.5μmol/L、5μmol/L、10μmol/L、20μmol/L、40μmol/L resveratrol of20%CBRH7919tk cells after48h, the inhibition rate was-0.43±1.77%,-0.85±0.58%,0.64±1.00%,9.75±1.45%,12.63±2.17%; A concentration of2.5μmol/L、5μmol/L、10μmol/L、20μmol/L、40μmol/L resveratrol combine with20%CBRH7919tk/GCV cells after48h, the inhibition rate was13.43±1.66%,14.02±1.92%,19.40±3.61%,24.15±2.84%,21.37±2.69%, Jin Shi Q values was1.31、1.56、1.87、1.29、1.01. In addition to40μmol/L combined with20%CBRH7919tk/GCV, other concentrations are synergies.(2) Western Blot detection impact of resveratrol on Cx26, Cx32, Cx43protein expression of CBRH7919cells.CBRH7919cells use resveratrol48h, Cx26protein expression is an upward trend; expression of Cx32protein are on the rise; Cx43protein expression is an upward trend.(3) Synergism of resveratrol in vivo system for CBRH7919TK/GCV Resveratrol100mg/kg tumor mass bigger than the untreated group, resveratrol150mg/kg tumor mass smallar than the untreated group, they have no statistical difference (P>0.05); Resveratrol100mg/kg joint GCV group, Resveratrol150mg/kg joint GCV group and GCV group tumor mass smaller than the untreated group, there were significant differences (P<0.05); Resveratrol100mg/kg joint GCV group compare with GCV group, no statistical difference (P>0.05), Resveratrol150mg/kg joint GCV group compare with GCV group, there were significant differences (P<0.05); Resveratrol100mg/kg group compare with Resveratrol100mg/kg joint GCV group, there were significant differences (P<0.05), Resveratrol150mg/kg group compare with Resveratrol150mg/kg joint GCV group, there were significant differences (P<0.05).Conclusion:Based on early experiments, live with herbal ingredient resveratrol further explore malignant melanoma in mice and rats with liver cancer two suicide gene system, found that resveratrol can increase the suicide gene system synergies, by adjusting the gap junction protein, to enhance the function of GJIC.