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浙江大学学报(医学版)  2015, Vol. 44 Issue (5): 486-492    DOI: 10.3785/j.issn.1008-9292.2015.09.03
专题报道     
SN-38与索拉非尼联合应用体外抗肝癌效果及其机制
徐力, 朱园润, 陈建, 杨晓春, 罗沛华
浙江大学药学院, 浙江杭州 310058
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
全文: PDF(2522 KB)  
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。

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    
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
收稿日期: 2015-05-15
CLC:  R963  
基金资助:

国家自然科学基金(81301892)

通讯作者: 罗沛华(1981-),女,博士,副教授,主要从事肿瘤药理学研究;E-mail:peihualuo@zju.edu.cn;http://orcid.org/0000-0002-6576-2052     E-mail: peihualuo@zju.edu.cn
作者简介: 徐力(1992-),男,博士研究生,主要从事肿瘤药理学研究;E-mail:lxu_zju@zju.edu.cn;http://orcid.org/0000-0003-3255-326X
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引用本文:

徐力等. SN-38与索拉非尼联合应用体外抗肝癌效果及其机制[J]. 浙江大学学报(医学版), 2015, 44(5): 486-492.
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.

链接本文:

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

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