The GJIC Mechanism Of Resveratrol On Suicide Gene Therapy For Liver Cancer

In recent years,with the development of molecular biology techniques and applications,Immunotherapy,gene therapy and other new methods,and a variety of therapies as joint become hot spot in the field of cancer therapy,The suicide gene therapy is thorough and has the application prospect tumor to treat the new measure,the world already had the multi-wrap suicide gene therapy system to carry on even already completedâ… -â…¢Time clinical trial Existing research found that almost all suicide gene system has bystander effect.Enhancing the bystander effect can lower the high rate of transfer requirements,reduce the quantity and toxicity of the carrier,enhance the mass destruction to tumor.Therefore,it has been an important strategy to enhance the bystander effect for gene therapy.Judging from the current study there are the main mechanisms of bystander effect:Gap Junctional Intercellular Communication(GJIC)-mediated,and apoptosis-mediated immune-mediated such as,etc.and GJIC is next to one of the most important mechanisms.Preliminary studies indicated that resveratrol under the conditions of low toxicity to the cells jointed HSV-tk/GCV system can significantly improve the GCV of tk⁺,tk^- cell hybrid of anti-rate,The mechanism may be enhanced through the GJIC cells and induction of apoptosis,cell cycle regulation,and it may be two mechanisms are working.Chinese medicine is a treasure house of natural medicines,and has advantages or potential in induceing apoptosis,enhancing GJIC or improving the immune function to cancer,at the same time,it has relatively few side effects,So searching for drugs to improve efficiency in Chinese medicine and to study its mechanism,the establishment of combining traditional Chinese and western medicine treatment is feasible and a good prospect.ObjectivesIn the topics we chosed for Chinese medicine resveratrol which can certainly enhance the bystander effect to suicide gene system,clarified its efficiency of the mechanism in order to establish a method of cancer gene therapy joint Chinese medicine and promote cancer gene therapy clinical application and lay the foundation for the development of new drugs.MethodsIn this paper,we cultured liver cancer cells HepG2 and CBRH7919,First, we made use of the method of MTT to observe the inhibition of resveratrol on HepG2 and calculate IC50,in order to identify drug concentration and time. Second,methods of SL/DT and FCM were used to observe the function of GJIC of HepG2 and CBRH7919,Western-blot was used to observe Cx43 and Cx32 expression of HepG2 and CBRH7919.Results1 The inhibition of resveratrol on HepG2There were no significant difference in all groups of cells before drug effects,however,the numbers of cells in 50μM group and 100μM group were observed that significantly reduced from the 24 hours;With the increase of incubation time,25μM group was also evident in cell growth inhibition.60 hours 10μM group has yet to observe significant growth inhibition,The adherent cells in other groups were observed gradually falling.10μM,25μM, 50μM,100μM of resveratrol on Hepg2 cells 60h,The survival rates were 90.8±4.0%(p=0.085＞0.05),73.6±4.8%(p=0.005＜0.01),40.5±3.9% (p=0.0007＜0.01),26.7±3.2%(p=0.0004＜0.01);IC50 for 43μM.This shows that there was certern inhibiting effect for resveratrol on HepG2 cell growth.2.SL/DT and FCM test showed that:resveratrol-treated groups,in contrast to control group,the proportion of double-positive(more positive noting GJ function better)was increased and there was dose-dependent,showing that resveratrol can improve the GJIC function of HepG2 and CBRH7919.3 Western blot test,which used polyclonal and monoclonal anti-Cx,have shown that:Resveratrol can significantly improve the expression of Cx32 of HepG2 and CBRH7919,moreover,there were concentration and time-dependent, oxidants H₂O₂ can inhibit the effect;but the expression of Cx43,which was not obvious in 24h,was increased in 48h. Conclusion:Based on the pre-active components,resveratrol can effectively improve the suicide gene on the inhibition of tumor cells and enhance the bystander effect,we study its mechanism for further,the results show that resveratrol can improve the expression of Cx and the function of GJIC.