HER2在人肺腺癌A549细胞培美曲塞继发性耐药中的作用研究  

The Role of HER2 in Secondary Drug-resistance in A549 Lung Adenocarcinoma Cells Induced by Pemetrexed

作　　者：侯宪鹏[1] 刘冰[1] 于壮[1] 刘相萍[1] 杨堃[1] 

机构地区：[1]青岛大学医学院附属医院,山东青岛266003

出　　处：《现代生物医学进展》2013年第18期3448-3451,共4页Progress in Modern Biomedicine

基　　金：吴阶平医学基金会资助(NO.320.6753.1219)

摘　　要：目的:通过体外诱导耐药建立肺腺癌A549细胞系的培美曲塞耐药细胞株,并进一步研究其耐药性发生与HER2/neu之间的关系。方法:通过高浓度反复间歇诱导法建立A549的培美曲塞耐药细胞系。CCK8法检测耐药性、绘制生长曲线,计算倍增时间;流式细胞术法检测细胞周期分布及细胞凋亡水平;RT-PCR法检测耐药株及亲本株HER2/neu的mRNA表达。结果:历经6个月建立了耐药指数为6.7倍的A549/PEM细胞系,其较亲本株的倍增时间延长(33.5±1.71h vs 27.1±1.15h);流式细胞术检测结果显示:与亲本株比较,A549/PEM处于G1期的细胞比例增加(66.03±1.61%vs 57.92±1.73%),S期细胞比例降低(30.05±1.25%vs 37.51±1.31%),差别均有统计学意义(P<0.05);在PEM作用24h后,A549与A549/PEM细胞凋亡率分别为23.52%和10.99%,差别有统计学意义(P<0.05)。RT-PCR检测结果示:A549/PEM较亲本株HER2/neu基因的mRNA表达水平增高(P<0.05)。结论:A549/PEM与亲本株的生物学特性有差别;肺腺癌培美曲塞继发性耐药的产生可能与HER2/neu表达上调有关。Objective：To establish a PEM-resistant lung adenocarcinoma cell line and to investigate the relationship between HER2/neu and PEM-resistance.Methods：The parental cell line A549 was treated repeatedly with high concentration of PEM.Growth inhibition was determined by the CCK8 assay to draw growth curves,and then the doubling time was calculated.The distribution of cell cycle and the level of cell apoptosis were evaluated by flow cytometry assay.Expression of HER2/neu was analyzed by quantitative real-time reverse transcriptase polymerase chain reaction.Results：A PEM-resistant lung adenocarcinoma cell line——A549/PEM was established,with resistance index of 6.7 times.Cell doubling time of A549/PEM and its parent strain was 33.5 and 27.1 hours,respectively,with a difference in terms of genetics between the two cell lines.Flow cytometry analysis showed G1 arrest in A549/PEM cell line and the apoptosis rate of A549 was higher than that of A549/PEM after 24-hours continuous exposing to PEM（23.52% Vs.10.99%）,with a difference in terms of genetics.A549/PEM expressed more HER2/neu mRNA than the parental cell（P0.05）,by polymerase chain reaction.Conclusion：The A549/PEM cell line showed some differences of the biological characteristics compared with the parental strain.HER2/neu expression appeared to be associated with resistance to PEM in lung adenocarcinoma.

关 键 词：非小细胞肺癌 HER2 NEU 培美曲塞 耐药 

分 类 号：R734.2[医药卫生—肿瘤]