半夏泻心汤含药血清对人胃癌耐药细胞株SGC7901/ADR化疗敏感性的作用

Study on the effect of Banxia Xiexin Decoction medicated serum on increasing human gastric cancer resistant cell line SGC7901/ADR chemosensitivity

目的 观察半夏泻心汤含药血清对人胃癌耐药细胞株SGC7901/ADR化疗敏感性的影响,并探讨其作用机制。方法 取对数生长期胃癌耐药细胞株SGC7901/ADR细胞,四甲基偶氮唑蓝(MTT)法检测半夏泻心汤含药血清对细胞耐药的逆转作用。取对数生长期细胞分为对照组、ADR组[23.72μmol/L(ADR IC_(50)的1/2浓度)ADR]、半夏泻心汤含药血清组[15.22μL/mL半夏泻心汤含药血清]、ADR+PI3K抑制剂(LY294002)组(23.72μmol/L ADR+20μmol/L LY294002)、ADR+半夏泻心汤含药血清组(23.72μmol/L ADR+15.22μL/mL半夏泻心汤含药血清)。给药72 h后,流式细胞术检测细胞周期和凋亡率;实时荧光定量聚合酶链反应和蛋白质印迹法检测细胞磷酸肌醇-3-激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)、细胞周期蛋白B1(cyclin B1)、p21、抗凋亡基因(Bcl-2)、促凋亡基因(Bax)信使核糖核酸(mRNA)和蛋白表达及p-PI3K、p-AKT、p-mTOR水平。结果 与单用ADR组细胞比,联合用药组细胞存活率下降(P<0.05或P<0.01);单用ADR IC_(50)值为47.44μg/mL,联合用药IC_(50)值为21.76μg/mL,两者比差异具有统计学意义(P<0.01);与ADR组比,ADR+LY294002组和ADR+半夏泻心汤含药血清组G_(2)/M期细胞显著增多(P<0.01)、细胞凋亡率显著增加(P<0.01)、PI3K、AKT、mTOR、cyclin B1、Bcl-2 mRNA和蛋白表达及p-PI3K、p-AKT、p-mTOR水平减少,p21、Bax mRNA和蛋白表达增加(P<0.05或P<0.01)。结论 半夏泻心汤含药血清可增加人胃癌耐药细胞株SGC7901/ADR化疗敏感性,可能与抑制PI3K/AKT/mTOR通路,抑制cyclin B1、Bcl-2表达,促进Bax和p21表达有关。

Objective To observe the effect of Banxia Xiexin Decoction medicated serum on the chemosensitivity of human gastric cancer resistant cell line SGC7901/ADR,and to explore the mechanism.Methods Logarithmic growth human gastric cancer resistant SGC7901/ADR cells were divided into control group and ADR group [23.7 μmol/L(1/2 concentration of ADR IC_(50)) ADR],Banxia Xiexin Decoction medicated serum group(15 μl/ml Banxia Xiexin Decoction medicated serum),ADR+PI3 K inhibitor(LY294002) group(23.7 μmol/L ADR+20 μmol/L LY294002),ADR+ Banxia Xiexin Decoction medicated serum group(23.7 μmol/L ADR+15 μl/ml Banxia Xiexin Decoction medicated serum).Seventy-two h after administration,cell cycle and apoptosis rate were detected by flow cytometry,and the mRNA expressions of phosphoinositol 3 kinase(PI3 K),protein kinase(AKT),mammalian target of rapamycin(mTOR),cyclin B1,p21,anti-apoptotic gene(Bcl-2) and pro-apoptotic gene(Bax),and the levels of p-PI3 K,p-Akt and p-mTOR were detected by real-time quantitative polymerase chain reaction and Western blotting.Results Compared with the ADR group alone,the cell viability in the combined treatment group decreased(P<0.05 or P<0.01).The IC_(50) value of cells treated with ADR alone was 47.4 μg/ml,and the IC_(50) value in the combined treatment group was 21.7 μg/ml(P<0.01).Cells in G_(2)/M phase of ADR+LY294002 group and ADR+Banxia Xiexin Decoction medicated serum group were increased(P<0.01) and the apoptosis rate was increased(P<0.01),mRNA and protein expressions of PI3 K,AKT,mTOR,cyclin B1,Bcl-2 and levels of p-PI3 K,p-Akt and p-mTOR were decreased,and the mRNA and protein expressions of p21 and Bax were increased(P<0.05 or P<0.01).Conclusion Banxia Xiexin Decoction medicated serum can increase chemosensitivity of human gastric cancer resistant cell line SGC7901/ADR,which may be related to inhibition of PI3 K/AKT/mTOR pathway,inhibition of cyclin B1,Bcl-2 expression,and promotion of Bax and p21 expression.