缺氧诱导因子-1α增加肿瘤细胞A549化疗药物耐受性的研究  

Hypoxia-inducible Factor-1alpha Suppresses Apoptosis and Increases Tolerance of Lung Cancer Cells to Chemotherapy

作　　者：张惠兰[1] 郦俊[1] 张珍祥[1] 徐永健[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院呼吸内科,武汉430030

出　　处：《华中科技大学学报（医学版）》2007年第5期578-580,584,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No30500224)

摘　　要：目的构建缺氧诱导因子-1α(hypoxia-inducible factor-1alpha,HIF-1α)基因的表达质粒,转染肺癌细胞系A549后,观察其对化疗药物5-氟尿嘧啶(5-Fu)敏感性的变化。方法采用RT-PCR方法扩增A549细胞内HIF-1α基因功能区片段,克隆至真核表达载体pcDNA3上,经脂质体LipofectamineTM2000转染A549细胞后,G418筛选。West-ern blot检测转染前后多药耐药蛋白(MDR1)的表达变化。加入化疗药物5-Fu(0.1mg/ml),用生长曲线观察A549细胞的生长情况,用流式细胞仪检测细胞凋亡指数,用Western blot检测Caspase3的活性变化。结果转染HIF-1α后,MDR1表达上调。转染HIF-1α组的细胞增殖抑制、凋亡指数、Caspase3的活性均低于单独加5-Fu组。结论HIF-1α能够增加肺癌细胞A549凋亡的阈值,从而增加细胞对化疗药物的耐受性。Objective The plasmid expressing hypoxia-inducible factor-1alpha（HIF-1α） was constructed and transfected into human lung cancer cells A549 in order to observe the changes of sensitivity of A549 to chemotherapy.Methods HIF-1α mRNA structure region was amplified by RT-PCR and inserted into plasmid pcDNA3.The expression plasmid pcDNA3/HIF-1α was transfected into A549 with Lipofectamine^TM2000.HIF-1α protein was detected by Western blot.After 5-Fu was added into A549 transfected with HIF-1α,the growth activity was measured by growth curve,apoptosis was detected by flow cytometry,and the levels of caspase3 MDR-1 were detected by Western blot.Results The constructed expression plasmid was analyzed with restriction enzymes and gel electrophoresis.Two DNA lanes at 2.55 kb and 5.4 kb respectively were found,which was consistent with that expected.The growth rate in 5-Fu group was inhibited as compared with control group and the apoptosis index and caspase3 activity were increased significantly.After HIF-1α was transfected into A549,the activity of MDR-1 was decreased,and the effect of 5-Fu was weakened.Conclusion HIF-1α can promote chemoresistance by increasing the activity of MDR1 and suppressing apoptosis of lung cancer cells A549 incurring 5-Fu.

关 键 词：缺氧诱导因子1Α 化疗 肺癌 

分 类 号：R562.2[医药卫生—呼吸系统]