【摘要】 目的:研究伊立替康的活性代谢物SN-38与索拉非尼联合作用于HepG-2和BEL-7402细胞株的抗肝癌效果及其相关机制。方法:利用磺酰罗丹明B显色法测定SN-38与索拉非尼单用或合用后HepG-2和BEL-7402细胞的存活率。利用PI染色结合流式细胞术及DAPI染色法检测细胞凋亡,同时利用蛋白质印迹法检测凋亡相关蛋白、DNA损伤标志蛋白的表达情况。结果:与单药比较,SN-38与索拉非尼合用对细胞的抑制作用增强,合用指数小于0.9。对照组、SN-38组、索拉非尼组、合用组HepG-2细胞凋亡率分别为4.25%±2.45%、28.95%±10.75%、3.49%±2.49%、53.19%±11.21%,合用组细胞凋亡率增加(与其他组比较均P<0.05)。同时合用组凋亡相关蛋白多聚二磷酸腺苷核糖聚合酶(PARP)、半胱氨酸天冬氨酸蛋白酶(Caspase)8、Caspase-3的蛋白酶切量以及p53蛋白、p21蛋白、DNA损伤标志蛋白磷酸化的组蛋白H2AX的表达量均增加。结论:在细胞水平上,SN-38与索拉非尼联合应用能够通过p53表达增加促进肝癌细胞凋亡,因此具有抗肝癌效果。更多还原

【Abstract】 Objective:To investigate the anticancer effect and its mechanism of SN-38 combined with sorafenib on hepatocellular cancer cell lines HepG-2 and BEL-7402.Methods:SRB colorimetry was employed to measure the viability of HepG-2 and BEL-7402 cells after the treatment of SN-38 with sorafenib.Propidium iodide flow cytometric assay and DAPI staining were used to evaluate the apoptosis of HCC cells.Western blotting was conducted to detect the expression level of apoptosis-related and DNA damage-related proteins.Results:SRB colorimetry showed the synergistic anticancer activities of SN-38 combined with sorafenib,with a combination index of <0.9.The apoptotic rates of HepG-2 cells in control,60 nmol/L SN-38,2.5 μmol/L sorafenib and combination groups were 4.25%±2.45%,28.95%±10.75%,3.49%±2.49%and 53.19%± 11.21%,respectively(P <0.05).Western blotting showed that the combination of these two drugs increased the enzymolysis of PARP,Caspase-8 and Caspase-3,and promoted the expression levels of p53,p21 and 7-H2 AX significantly.Conclusion:SN-38 and sorafenib have synergistic anticancer activity on hepatocellular carcinoma cells in vitro with the augmentation of apoptosis.更多还原