芥子碱硫氰酸盐增强HepG2细胞对吉西他滨敏感性的作用

目的探讨芥子碱硫氰酸盐增强人肝癌细胞HepG2对吉西他滨敏感性的作用及分子机制。方法应用计算机分子对接技术分析芥子碱硫氰酸盐结合多耐药性相关蛋白ABCB1和ABCG2的能力,罗丹明123实验检测芥子碱硫氰酸盐对HepG2细胞外排药物能力的影响,CCK-8、克隆形成实验、流式细胞术分别检测芥子碱硫氰酸盐联合吉西他滨对HepG2细胞增殖、克隆形成、凋亡的影响,Western blot实验检测芥子碱硫氰酸盐联合吉西他滨对HepG2细胞中ABCB1、ABCG2、活化半胱天冬酶-8(Cleave caspase-8)、B淋巴细胞瘤-2基因(Bcl-2)、凋亡相关基因(Bax)表达的影响。结果芥子碱硫氰酸盐可稳定结合多耐药性相关蛋白ABCB1、ABCG2,降低HepG2细胞外排药物的能力,增强吉西他滨对HepG2细胞增殖和克隆形成的抑制作用(P<0.05)及吉西他滨诱导HepG2凋亡的能力(P<0.05),抑制ABCB1、ABCG2蛋白表达,增强吉西他滨诱导的Bcl-2表达减少及Cleave caspase8、Bax表达增加(P<0.05)。结论芥子碱硫氰酸盐能靶向抑制多耐药相关蛋白ABCB1和ABCG2,增强肝癌HepG2细胞对吉西他滨的敏感性。

Effects of sinapine thiocyanate in promoting the sensitivity of HepG2 cells to gemcitabine

AIM To detect the effect of sinapine thiocyanate on promoting the sensitivity of human hepatocellular carcinoma(HCC)cell HepG2 to gemcitabine,and the molecular mechanism as well.METHODS The ability of sinapine thiocyanate to bind with multi-drug resistance related proteins ABCB1 and ABCG2 was analyzed using computer molecular docking technique;rhodamine 123 was used to detect the effect of sinapine thiocyanate on the drug efflux ability of HepG2 cells;the effect of the combinative use of sinapine thiocyanate and gemcitabine was tested in terms of the proliferation,colony formation and apoptosis of HepG2 cells by CCK-8,colony formation assay and flow cytometry,and the expression of ABCB1,ABCG2,caspase-8,Cleave caspase-8,Bcl-2 and Bax in HepG2 cells by Western blot.RESULTS Stably binded with multi-drug resistance related proteins,sinapine thiocyanate compromised ABCB1 and ABCG2 expression,reduced the drug efflux in HepG2 cells;enhanced gemcitabine-induced inhibition of HepG2 cell proliferation and colony formation(P<0.05)and HepG2 cell apoptosis(P<0.05),reduction of Bcl-2 expression and the increase of cleave caspase8 and Bax expression as well(P<0.05).CONCLUSION Sinapine thiocyanate promotes the sensitivity of human HCC cell HepG2 to gemcitabine via targeting multi-drug resistance related proteins ABCB1 and ABCG2.