摘要
目的探讨单宁酸(TA)与顺铂(CDDP)协同抗肝癌细胞HepG2的作用及其内质网应激(ERS)通路的激活情况。方法将人肝癌细胞HepG2分为对照组、TA组、CDDP组、TA+CDDP组(联合组),并在体外培养24h。MTT法测定药物对人肝癌细胞HepG2的生长抑制作用;药物联合作用指数、药物剂量降低指数和等效图解法分析两药物之间药效学的协同作用;4′6二脒基-2-苯基吲哚(DAPI)染色观察细胞核变化;实时荧光定量PCR(q-RT-PCR)和免疫印迹检测ERS标志分子葡萄糖调节蛋白(GRP)78、GRP94的水平。结果 TA和CDDP对HepG2细胞生长均呈剂量依赖性抑制作用,其半数有效浓度(IC50)分别为360.00μmol/L和1.80μg/mL;用180.00μmol/L TA和0.90μg/mL CDDP联合处理肝癌HepG2细胞后,能够加强对细胞生长的抑制作用;TA和CDDP具有协同抑制肝癌细胞生长的作用。与TA、CDDP组比较,联合组细胞皱缩、变圆加剧且凋亡细胞增多,细胞核固缩、边缘不规则、致密浓染及核裂解碎片增多;细胞GRP78、GRP94的表达均上升。结论 TA能够增强CDDP的抗肝癌HepG2细胞作用,该协同作用的机制可能与激活ERS通路有关。
Objective To investigate the synergistic effects of tannic acid（TA） and cis-dichlorodiamine platinum（CDDP） on hepatocellular carcinoma HepG2 cells and its activation situation of endoplasmic reticulum stress（ERS） pathway. Methods Human hepatocellular carcinoma HepG2 cells were divided into the control group,TA group,CDDP group and TA＋CDDP group, and were cultured in vitro for 24 h. The growth inhibitory effect of medication on HepG2 cells was detected by MTT assay. The pharmacody- namics synergistic effect between the two drugs was analyzed by the drug interaction index,drug dose reduction index and equiva- lent graphical method. The nucleus changes were observed by DAPI staining. Real time fluorescent quantitative PCR（q-RT-PCR） and Western blot were used to detect the levels of ERS markers glucose regulated protein （GRP）78 and GRP94. Results TA and CDDP had dose-dependent growth inhibition effect on HepG2 cells,their median effective concentrations（ICs0）were 360.00 /μmol/ L and 1.80 μg/mL respectively. The combination treatment of 180.00 μmol/L TA and 0.90 μg/mL CDDP on HepG2 cells could enhance the inhibitory effect on cell growth. Ta and CDDP had synergistic effect for inhibiting hepatocellular carcinoma cells growth. Compared with the TA group and CDDP group, cell shrinkage and rounding were accelerated in the combined group,apop- totic cells were increased, nuclear had pyknosis,irregular edge and dense staining, nuclear fragmentations were increased and the ex- pressions of GRP78 and GRP94 were up-regulated. Conclusion TA can enhance the effect of CDDP on anti-hepatic carcinoma HepG2 cells,and the synergy mechanism may be related to the activation of ERS pathway.
出处
《重庆医学》
CAS
2018年第5期594-597,600,共5页
Chongqing medicine
基金
贵州省科技厅资助项目[黔科合J字(2013)2319号
黔科合LH字(2014)7548号]
贵州省教育厅特色药用资源研发创新团队[黔科合人才团队字(2013)15号]
合肥医学院博士启动基金(F-655)
合肥医学院遗传学扶持学科经费(2012-2016)
关键词
单宁酸
顺铂
药物协同作用
肝肿瘤
细胞凋亡
内质网应激
tannic acid
cis-dichlorodiamine platinum
drug synergism
liver neoplasms
apoptosis
endoplasmic＂ reticulumstress
