奥美拉唑逆转人肺腺癌耐顺铂细胞株A549/DDP耐药性的研究  

Omeprazole reverses the drug resistance in cisplatin-resistant human lung adenocarcinoma cell line A549/DDP

作　　者：杨列军[1] 姜达[1] 

机构地区：[1]河北医科大学第四医院肿瘤内科,石家庄050011

出　　处：《肿瘤》2010年第11期924-928,共5页Tumor

摘　　要：目的:探讨液泡ATPase[vacuolar(H+)-ATPase,V-ATPase]非特异性抑制剂奥美拉唑(omeprazole,OME)逆转人肺腺癌耐药细胞株A549/DDP的耐药性及可能的作用机制。方法:以非细胞毒性浓度4μg/mL的OME预处理A549/DDP细胞24 h,再用2μg/mL顺铂(cisplatin,DDP)处理A549/DDP细胞。MTT法检测细胞的增殖抑制率,激光扫描共聚焦显微镜(laser scanning confocal microscope,LSCM)法间接检测细胞内pH值的变化,FCM法检测凋亡相关蛋白Bcl-2和PTEN的表达,RT-PCR法检测VATPase、Bcl-2和PTEN mRNA的表达。结果:OME具有逆转A549/DDP细胞耐药性的作用,逆转倍数为1.45(P<0.01);OME预处理后A549/DDP细胞内pH值明显降低,Bcl-2蛋白表达下降,PTEN蛋白表达增加(P<0.01)。V-ATPase在亲本A549及A549/DDP细胞中相对表达量分别为0.88±0.00和0.99±0.00(P<0.01);A549/DDP细胞中V-ATPase在有无OME预处理的情况下的相对表达量分别为1.09±0.00和1.05±0.02,差异无统计学意义。Bcl-2 mRNA相对表达量下降(P<0.01),PTEN mRNA相对表达量增加(P<0.01)。结论:V-ATPase在A549/DDP细胞中高表达,OME能部分逆转A549/DDP耐药性,其作用机制可能与上调PTEN和下调Bcl-2表达有关。Objective:To study the effects of omeprazole(OME),a nonspecific inhibitor of vacuolar(H+)-ATPase,in reversing the cisplatin resistance of human lung adenocarcinoma cell line A549/DDP and elucidate the possible mechanism.Methods:A549/DDP were pretreated with OME at non-toxic concentration of 4 μg/mL for 24 h and then incubated with cisplatin(DDP) 2 μg/mL.Cell proliferation was measured with MTT assays.The intracellular pH was detected using laser scanning confocal microscopy(LSCM).The intracellular Bcl-2 and PTEN protein expressions were measured using flow cytometry(FCM).The mRNA expressions of V-ATPase,Bcl-2,and PTEN were detected using semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).Results:OME reversed the drug resistance of A549/DDP cells.The reversal multiple was 1.45(P<0.01).LSCM indirectly showed that the intracellular pH was decreased after OME pretreatment.The expression of Bcl-2 in A549/DDP cells was decreased(P<0.01),but the expression of PTEN was increased(P<0.01).The relative expressions of V-ATPase mRNA were 0.88±0.00 and 0.99±0.00 in A549 and A549/DDP cells,respectively(P<0.01).The relative expressions of V-ATPase mRNA of A549/DDP cells were 1.09±0.00 and 1.05±0.02 in OME-treated group and OME-untreated group.RT-PCR assay showed that pretreatment with OME for 24 h inhibited the relative expressions of Bcl-2 mRNA in A549/DDP cells and increased the PTEN mRNA expression(P<0.01).Conclusion:The V-ATPase mRNA is over-expressed in A549/DDP cells.OME partly reverses the drug resistance of A549/DDP cells.The action mechanism may be related with up-regulation of PTEN expression and down-regulation of Bcl-2 expression.

关 键 词：奥美拉唑 人肺腺癌 顺铂 耐药细胞株 耐药性 cell line lung adenocarcinoma human A549/DDP细胞 V-ATPase 表达量 Bcl-2蛋白表达 激光扫描共聚焦显微镜 PTEN蛋白表达 confocal microscope 预处理 OME 作用机制 检测 细胞内 

分 类 号：R73[医药卫生—肿瘤]