摘要
目的 探讨p38MAPK磷酸化对HepG2细胞耐药的影响及其意义.方法 采用浓度递增法,建立动态HepG2/顺铂（CDDP）耐药模型,Western blot检测磷酸化p38的表达;予以p38MAPK特异性抑制剂SB203580分别处理24、48、72 h后,流式细胞仪动态分析HepG2/CDDP耐药细胞周期、四甲基偶氮唑盐法检测顺铂对HepG2/CDDP耐药细胞的半数药物抑制浓度（IC50）、Western blot检测HepG2/CDDP耐药细胞凋亡分子Bcl-2、Bax及P-gp蛋白的表达.动态耐药模型磷酸化P38的检测行单因素方差分析; SB203580处理后计量资料行3×3析因设计方差分析,率的比较采用x2检验.结果 成功建立HepG2/CDDP耐药细胞株,其对CDDP的IG50值与对照HepG2细胞差异具有统计学意义（t=99.30,P〈0.01）;随着HepG2/CDDP耐药细胞株耐药表型的增强,P38MAPK活化水平逐渐增强（F=69.39,P〈0.01）.P38MAPK特异性抑制剂SB203580可逐渐降低HepG2/CDDP对顺铂的IC50值（F=2350,P〈0.01）、G0/Gl期细胞比例（x2=520,P〈0.01）、Bcl-2/Bax表达比值（F=83.8,P〈0.01）及细胞膜药物转运相关蛋白P-gP的表达（F=107,P〈0.01）.结论 P38活化水平随着肝癌耐药细胞株耐药表型的增强而逐渐升高;抑制P38的活化可有效逆转肝癌HepG2/CDDP耐药细胞株的耐药性.
Objective To investigat the effectiveness of p38MAPK activity during drug resistance against HepG2. Methods HepG2/CDDP kinetic anti-cancer drug resistance model was constructed using anti-cancer drug inducing method and treated with P38MAPK inhibitor SB203580 for 24, 48 and 72 hours respectively. Flow cytometry （FCM） was used to detect the cell cycle distribution. The IC50 of cisplatin was determined by MTT method in vitro. The expressions of P-P38, P-gp, Bcl-2 and Bax were examined by Western blot. Retults The HepG2/CDDP kinetic drug resistance model was successfully established. The expression of P-P38 increased with the increasing drug resistance against HepG2 cells. The models treated with SB203580 could gradually elevate the sensitivity of HepG2/CDDP to cisplatin, block the detention of cell cycle, up-regulate the expression of Bax and down-regulate the expressions of Bcl-2 and P-gp. Conclusion The expression of P-P38 increased with the increasing drug resistance against hepatocellular carcinoma cells. Supressing the activation of P38 could reverse the drug resistance phenotype against hepatocellular carcinoma cells.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2010年第8期604-608,共5页
Chinese Journal of Hepatology
基金
重庆市教育委员会科学技术研究项目（0200050174KJ090303）
关键词
癌
肝细胞
耐药性
细胞周期
P38
Carcinoma,hepatocellular,Drug resistance
Cell cycle
P38
