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Journal of ZheJiang University(Medical Science)  2015, Vol. 44 Issue (5): 486-492    DOI: 10.3785/j.issn.1008-9292.2015.09.03
    
Anticancer effect of SN-38 combined with sorafenib on hepatocellular carcinoma in vitro and its mechanism
XU Li, ZHU Yuan-run, CHEN Jian, YANG Xiao-chun, LUO Pei-hua
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China
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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, 2.5μmol/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 γ-H2AX significantly. Conclusion:SN-38 and sorafenib have synergistic anticancer activity on hepatocellular carcinoma cells in vitro with the augmentation of apoptosis.



Key wordsLiver neoplasms/drug therapy      Carcinoma, hepatocellular/pathology      Benzenesulfonates/therapeutic use      Camptothecin/analogs &      derivatives      Camptothecin/therapeutic use      Drug therapy, combination      Apoptosis      Genes, p53     
Received: 15 May 2015     
CLC:  R963  
Cite this article:

XU Li, ZHU Yuan-run, CHEN Jian, YANG Xiao-chun, LUO Pei-hua. Anticancer effect of SN-38 combined with sorafenib on hepatocellular carcinoma in vitro and its mechanism. Journal of ZheJiang University(Medical Science), 2015, 44(5): 486-492.

URL:

http://www.zjujournals.com/xueshu/med/10.3785/j.issn.1008-9292.2015.09.03     OR     http://www.zjujournals.com/xueshu/med/Y2015/V44/I5/486


SN-38与索拉非尼联合应用体外抗肝癌效果及其机制

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。


关键词: 肝肿瘤/药物疗法,  癌,肝细胞/病理学,  苯磺酸盐类/治疗应用,  喜树碱/类似物和衍生物,  喜树碱/治疗应用,  药物疗法,联合,  细胞凋亡,  基因, p53 
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