五味子甲素协同吉西他滨抑制肝癌细胞HepG2增殖

目的探究五味子甲素(deoxyschizandrin)联合吉西他滨(gemcitabin,GEM)对肝癌细胞HepG2增殖的影响及其可能的作用机制。方法CCK8法和克隆平板实验检测五味子甲素单药、GEM单药及联合用药对肝癌细胞HepG2增殖能力的影响;流式细胞术检测单药组及联合用药组中HepG2细胞的凋亡比例;Western blot法检测各组细胞中BCL2、BAX、pro-caspase3、cleaved-caspase3、pro-caspase9、cleaved-caspase9、β-catenin和TCF-4的表达。结果五味子甲素、GEM及联合用药对细胞HepG2的增殖均具有抑制作用,促进其凋亡,且联合用药组对HepG2细胞的作用明显强于单药组(P<0.05);Western blot结果表明,与对照组相比,五味子甲素、GEM和联合用药对pro-caspase3、pro-caspase9表达无明显影响,显著减少BCL2、β-catenin及TCF-4蛋白的表达(P<0.05),上调BAX、cleaved-caspase3、cleaved-caspase9蛋白的表达(P<0.05)。其中,联合用药比单药更为显著(P<0.05)。结论五味子甲素协同GEM抑制肝癌细胞HepG2中β-catenin/TCF-4通路,进而减少细胞增殖,诱导其凋亡。

Synergy effect of deoxyschizandrin and gemcitabine on proliferation of HepG2 human hepatocellular carcinoma cells

Aim To investigate the synergy effects of deoxyschizandrin and gemcitabine(GEM)on the proliferation of HepG2 human hepatocellular carcinoma cells in vitro and the underlying mechanism.Methods CCK8 method and colony formation assay were used to detect the effects of deoxyschizandrin monotherapy,GEM monotherapy and combination therapy on the proliferation of HepG2 cells.Flow cytometry was used to detect the change of apoptosis rate of HepG2 cells after treatment with single drug or the combination use of two drugs.Western blot was performed to detect the expression of BCL2,BAX,pro-caspase3,caspase3,pro-caspase9,caspase9,β-catenin and TCF-4.Results Deoxyschizandrin,GEM and combination group significantly inhibited the proliferation of HepG2 cells and promoted cell apoptosis.The effects of the combination group on HepG2 cells were significantly stronger than those of single-drug groups(P<0.05).Western blot results showed the expression of pro-caspase3 and pro-caspase9 was changed slightly within deoxyschizandrin,GEM and combination groups compared with that of normal control,while the expression of BCL2,β-catenin and TCF-4 protein expression was down-regulated significantly(P<0.05).The expression of BAX,cleaved-caspase3 and cleaved-caspase 9 protein increased significantly after treatment with deoxyschizandrin,GEM and combination group(P<0.05).Specially,the increasing effect of the expression of the protein in combined group was more significant than that of single drug groups(P<0.05).Conclusions The combination of deoxyschizandrin and GEM significantly inhibited the proliferation of HepG2 cells and induced cell apoptosis,as well as suppressed theβ-catenin/TCF-4 pathway.