摘要
目的探讨藤黄酸(GA)对人胃癌SGC7901/DDP细胞顺铂敏感性的影响及其分子机制。方法采用顺铂(DDP)浓度梯度递增法构建人胃癌顺铂耐药株SGC7901/DDP细胞,采用细胞计数盒-8(CCK-8)法检测藤黄酸和顺铂对SGC7901/DDP细胞的毒性作用,采用Chou-Talalay中效分析法定量评价藤黄酸和顺铂的联合作用效果,采用流式细胞术检测细胞凋亡,采用Western blotting方法检测Bcl-2、Bax、Survivin、多药耐药相关蛋白2(MRP2)、磷酸化氨基末端蛋白激酶(p-JNK)(Thr183/Tyr185)和JNK的蛋白水平。结果藤黄酸与顺铂各自单独作用48 h的IC50分别为2.94μmmol/L和39.76μmmol/L;当抑制率超过20%时,两者联合应用呈协同效应;藤黄酸可协同增强顺铂诱导的细胞凋亡(P<0.05),下调Survivin和MRP2蛋白水平(P<0.05),上调Bax蛋白水平(P<0.05),抑制JNK磷酸化(P<0.05);JNK特异性抑制剂SP600125可下调MRP2蛋白水平(P<0.05)。结论藤黄酸可增强人胃癌SGC7901/DDP细胞对顺铂的敏感性,这可能与藤黄酸通过抑制JNK信号通路下调MRP2蛋白表达,以及上调Bax蛋白表达和下调Survivin蛋白表达有关。
Objective The aim of this study is to investigate the effect of gambogic acid （GA） on the sensibility of human gastric carcinoma SGC7901/DDP cells to cisplatin （DDP） and its possible mechanism. Methods The drug- resistant SGC7901/DDP cells were established by stepwise exposure to DDP. CCK-8 assay was employed to detect the cytotoxic effect of GA and DDP on SGC7901/DDP ceils, and the combined effect of GA and DDP was analyzed by Chou- Talalay method . The cell apoptosis was studied by flow cytometry. Western blotting was performed to detect the protein levels of Bcl-2, Bax, Survivin, multidrug resistance-associated protein 2 （MRP2） , phosphorylated c-Jun N-terminal kinase （p-JNK） （Thr183/Tyr185） and JNK. Results The ICso values of GA and DDP were 2.94 ixmol/L and 39.76 Ixmol/L after 48 hours treatment respectively, and the combined both drugs improved the antitumor effects on SGC7901/DDP cells. GA enhanced eisplatin-induced apoptosis （ P 〈 0. 05） , down-regulated Survivin and MRP2 and up-regulated max at the protein level （P 〈 0. 05 ）, and inhibited phosphorylation of JNK in SGC7901/DDP cells （ P 〈 0. 05 ）. SP600125, a specific inhibitor of JNK, declined the protein level of MRP2 （P 〈 0. 05）. Conclusion GA may enhance the sensibility of SGC7901/DDP cells to eisplatin by down-regulating MRP2 as a result of inactivation of the JNK signaling pathway, and up- regulating max and down-regulating Survivin.
作者
仝雷
王丽君
袁磊
TONG Lei;WANG Li-jun;YUAN Lei(Department of Medical Morphology;Department of Medical Function, Zhengzhou University of Industrial Technology, He＇ nan Xinzheng 451150, China;Key Laboratory of Medical Bioengineering, Luohe Medical College, He＇ nan Lttohe 462002, China)
出处
《解剖学报》
CAS
CSCD
北大核心
2018年第3期337-341,共5页
Acta Anatomica Sinica
基金
河南省科技厅科技发展计划(142102310203)
