EGFR抑制剂AG1478增强奥沙利铂诱导HCT116细胞凋亡的作用

EGFR inhibitor AG1478 enhances oxaliplatin induced apoptosis in HCT116 cells

目的探究酪氨酸激酶抑制剂AG1478(Tyrphostin AG1478,AG1478)联合奥沙利铂(oxaliplatin,OXA)对结直肠癌细胞HCT116凋亡的作用。方法MTT法检测AG1478联合OXA对HCT116细胞增殖的影响;实时荧光定量PCR法(qRT-PCR法)检测p53、caspase-3、Bcl-2、Bax的mRNA表达水平;Western blot法检测自噬和凋亡相关蛋白(p53、caspase-3、cleaved-caspase 3、Bcl-2、Bax、p62、LC3、IL-6)的表达水平。结果OXA和AG1478均抑制人结直肠癌细胞HCT116增殖(P<0.01),加药48 h后,IC_(50)分别为23.47和26.58 mg·L^(-1)。与OXA单药组比较,AG1478与OXA联合用药组明显抑制HCT116细胞的增殖能力(P<0.01);联合用药组LC3、IL-6蛋白表达明显高于对照组和单用药组,p62明显下调(P<0.01);联合用药组p53、caspase-3的mRNA和蛋白表达水平升高,cleaved-caspase 3蛋白表达水平升高,Bax/Bcl-2比值明显高于对照组(P<0.01)。结论AG1478与OXA联合用药,能协同增强对HCT116细胞的抑制作用,进而诱导凋亡。

Aim To investigate the effect of tyrosine kinase inhibitor AG1478 combined with oxaliplatin(OXA)on apoptosis of colorectal cancer HCT116 cells.Methods MTT assay was used to measure the effect of AG1478 combined with OXA on proliferation of HCT116 cells.RT-qPCR was used to detect the mRNA expression levels of p53,caspase-3,Bcl-2 and Bax.Western blot was used to detect the proteins expression of p53,caspase-3,cleaved-caspase 3,Bcl-2,Bax,p62,LC3 and IL-6.Results Both OXA and AG1478 inhibited the proliferation of HCT116(P<0.01).IC_(50) was 23.47 mg·L^(-1) and 26.58 mg·L^(-1) for 48 hours.Compared with OXA treatment,the AG1478 and OXA combination group significantly inhibited the proliferation capacity of HCT116 cells(P<0.01).The combination group significantly up-regulated LC3 and IL-6 protein expression and down-regulated p62(P<0.01).The combination group significantly increased the mRNA and protein expression levels of p53,caspase-3.The protein expression levels of cleaved-caspase 3 were promoted,and the ratio of Bax/Bcl-2 was significantly higher than that of the control group(P<0.01).Conclusions AG1478 with OXA synergistically enhance the proliferation inhibition and then induce apoptosis of HCT116 cells.