健脾理气中药对肝癌端粒酶活性的影响

目的：在体内和体外观察健脾理气中药对端粒酶活性的影响，探讨其抗肿瘤治疗的作用机制。方法：采用TRAP方法，观察H22肝癌荷瘤小鼠在服用健脾理气中药后肿瘤端粒酶活性的变化，以及健脾理气中药对体外培养的人肝癌细胞株SMMC-7721端粒酶活性的影响。结果：体内实验显示中药组端粒酶的活性明显低于对照组(P=0．038)。体外实验表明，健脾理气冲剂对SMMC-7721细胞的端粒酶活性有抑制作用。结论：对肿瘤细胞端粒酶活性的抑制可能是健脾理气中药抗肿瘤作用机制的一部分。

Inhibition of telomerase activity by JianPiLiQi formula in liver cancer.

Objective:JianPiLiQi method is one of important parts in cancer especially the liver cancer clinical treatment process.Telomerase,an RNA dependent DNA polymerase that used by cancer cells to solve the continued shorted telomeres caused by end replication question.Therefore the cancer cell achieved an unlimited replicated age by this mechanism but somatic cell not.Activation and up regulation of telomerase are considered to be an important role in the genesis and development of cancer,and it becomes a hot point in cancer research.The aim is to investigate the anti cancer mechanism of JianPiLiQi formula by observing the inhibition of telomerase activity in vivo and in vitro.Methods:The JianPiLiQi formula we used in our study is made up of ten Chinese traditional medicines:amyda sinensis,micromelum falcatum,crataegus pinnatifida,codonopsis pilosula,poncirus trifoliata,akebia quinata,oldenlandia diffusa,poria cocos,et al.In vivo,treated with JianPiLiQi formula from the second day after inoculated the HAC mouse liver cancer cell into armpit of mice.The administration of JianPiLiQi formula is 30 g/kg and mice were killed at 14 th day.In vitro,the IC 50 concentrations in SMMC-7721 human liver cell mensurated by MTT method at third and firth days are 47 mg/ml and 32 mg/ml respectively.The drug concentration we used in cell culture is 40 mg/ml.The culture times are 1,3,5 days and removed drug followed 2 days culture respectively.Telomerase activity detection used Telomeric Repeat Amplification Protocol(TRAP)combined with polyacrlamide and agarose gels eletrophresis(PAGE) silver staining.Results:In vivo,JiPiLIQi formula have a 38% cancer inhibition rate,and the cancer telomerase activity in treatment group obvious lower than control ( P =0 038);in vitro,JianPiLiQi formula inhibited telomerase activity in SMMC 7721 liver cancer cell,the inhibited effect is associated with time.The activity of enzyme is no obvious inhibition in first day and medial inhibition in third days,but in firth day the inhibition is obvious.In addition,telomerase activity can resume in some degree when removed drug and followed 2 days culture.Conclusions:Our study showed JianPiLiQi formula is a good method in liver cancer treatment,telomerase activity inhibition should be one part of its anti cancer mechanisms.And the anti cancer effect by JianPiLiQi formula is inhibition but not kill.