摘要
探讨eIF4E对厄洛替尼耐药细胞株细胞增殖的影响及作用机制.在配对的厄洛替尼敏感和耐药细胞株(HCC827-EP、HCC827-ER)上,采用实时定量PCR检测mRNA水平的变化,Western Blot检测蛋白水平的变化;通过SRB检测细胞的生长和药物的抑制率的变化.以及用脂质体转染eIF4E的小干扰RNA,用上述方法检测多种信号分子mRNA、蛋白水平的变化,及对细胞生长和对药物抑制率的影响.结果显示,与HCC827-EP相比,eIF4E在HCC827-ER上高表达;HCC827-ER转染eIF4E siRNA的细胞生长速度明显减慢,从第3 d起差异具有统计学意义(P<0.05),在相同浓度的厄洛替尼作用下,转染eIF4E siRNA的细胞具有更高的抑制率,差异具有统计学意义(P<0.05).结论:下调eIF4E的表达能够增强厄洛替尼对NSCLC的抑制作用,eIF4E的高表达参与了厄洛替尼获得性耐药的产生.
To observe the effects of eIF4E on the proliferation of erlotinib resistant cells and mechanism. Paired erlotinib sensitive and resistant HCC827-EP and HCC827-ER cells were used. Protein and mRNA expression levels were detected by Western blot and quantitative RT-PCR. The 5 ds growth curve and inhibition effects of erlotinib were determined by SRB assay. Downregulation of eIF4E expression were achieved by transfection of cells with siRNA targeting eIF4E using lipofectamine,and nonsense siRNA were used as control. eIF4E expression was increased in HCC827-ER cells compared with HCC827-EP cells. And downregulation of eIF4E expression inhibited the growth of HCC827-ER cells（P〈0. 05）and sensitized their reponses erlotinib possibly through downregualtion of eIF4E expression（P〈0. 05）. Downregulation of eIF4E expression inhibited the growth of NSCLCs and enhanced the inhibitory effects of erlotinib.
出处
《南京师大学报（自然科学版）》
CAS
CSCD
北大核心
2014年第2期91-95,共5页
Journal of Nanjing Normal University(Natural Science Edition)
基金
国家自然科学基金(30873099
81102458
811172004)
基础医学院优势学科
校引进人才启动基金
