摘要
背景与目的热休克蛋白(heatshockprotein,Hsp)90是Hsps家族中的重要成员,其高表达与肿瘤的发生、发展及化疗耐药密切相关。本研究旨在探讨替普瑞酮(geranylgeranylacetone,GGA)作用下人肺癌细胞SPCA-1及H446中Hsp90的表达水平变化及与肺癌细胞对顺铂化疗耐药的相关性。方法将细胞分成实验组与对照组,分别用含有不同浓度(0μM,10μM,50μM,100μM,500μM,1,000μM)的诱导剂GGA处理6h。采用免疫荧光细胞化学及West-ernblot方法检测各组细胞中Hsp90在蛋白水平的表达;应用MTT法测定细胞在化疗药物顺铂作用下生存率,并分析Hsp90的表达在两种肺癌细胞对顺铂耐药中的作用。结果 SPCA-1及H446实验组细胞中Hsp90的表达水平均明显高于相应的对照组细胞,且与GGA具有一定的浓度依赖性。MTT显示两种实验组细胞对顺铂的生存率均明显高于相应的对照组细胞,亦呈一定的GGA浓度依赖性。结论 GGA可以诱导人肺癌细胞SPCA-1及H446中的Hsp90的表达上调,且其表达水平都与GGA具有一定的浓度依赖性。Hsp90高表达的细胞对顺铂的生存率明显高于低表达的细胞,表明Hsp90的表达上调与肺癌细胞对顺铂的耐药有一定的相关性。
Background and objective Hsp90 is a major molecular chaperone, which overexpression is involved in oncogenesis, development and drug resistance in many human cancers. The aim of this study is to investigate the relationship between the GGA-induced overexpression of Hsp90 and chemoresistance to Cisplatin in SPCA-1 and H446 cell line. Methods The protein expressions of Hsp90 induced by GGA at different concentrations were analyzed by Immunofluorescence and Western blotting. Cells survival to Cisplatin was determined using the MTT assays. The effect of Hsp90 expression on the drug resistance to Cisplatin in two Cell Lines was analyzed. Results Compared with the respective control cells, Hsp90 expressions in both experimental cell lines were up-regulated obviously, exhibiting a dose-dependent manner to GGA. MTT assays revealed that the IC50s of cisplatin also showed a substantial elevation for the experimental cells of SPCA-1 and H446, and this elevation was also associated with GGA concerntration. Conclusion GGA is effective for the induction of Hsp90 in the SPCA-1 and H446 cell line. Up-regulated Hsp90 is associated with the chemoresistance to Cisplatin in SPCA-1and H446 cells.
出处
《中国肺癌杂志》
CAS
2011年第6期472-477,共6页
Chinese Journal of Lung Cancer
