靶向阻断LRP提高肺腺癌细胞对紫杉醇的敏感性  

Targeted blocking LRP increases chemotherapeutic sensitivity to taxol in lung carcinoma cells

作　　者：张善锋[1] 王志举[2] 崔景彬[3] 张艳杰[2] 张文瑾[2] 

机构地区：[1]郑州大学基础医学院生物化学与分子生物学系,河南郑州450001 [2]郑州大学基础医学院生理学与神经生物学系,河南郑州450001 [3]郑州大学基础医学院基础肿瘤教研室,河南郑州450001

出　　处：《基础医学与临床》2016年第3期370-374,共5页Basic and Clinical Medicine

基　　金：河南省高校科技创新人才支持计划(2012HASTIT004)

摘　　要：目的利用小干扰RNA(siRNA)技术沉默肺腺癌细胞肺耐药相关蛋白基因(LRP),探讨其对肺腺癌紫杉醇耐药株(A549/TXL20)紫杉醇(TXL)敏感性的影响。方法逐步增加药物浓度法建立A549紫杉醇耐药细胞株(A549/TXL20),用小干扰RNA技术沉默LRP在A549/TXL20细胞中的表达,以MTT法检测紫杉醇对A549/TXL20细胞的半数抑制浓度(IC50);以q PCR检测细胞中LRP mRNA的表达,Western blot检测细胞中LRP蛋白的水平,裸鼠腋窝皮下注射转染后的A549/TXL20细胞,建立裸鼠移植瘤模型,观察LRP靶向siRNA对人肺腺癌耐药细胞株A549/TXL20移植瘤耐药的影响。结果肺腺癌紫杉醇耐药株(A549/TXL20)对紫杉醇的敏感性明显增强(P<0.01),A549/TXL20细胞中LRP mRNA及蛋白表达显著增加(P<0.01);siRNA沉默A549/TXL20细胞LRP基因后,与空白组和空质粒组比较,LRP mRNA及蛋白表达均被抑制(P<0.01);小干扰RNA可提高荷瘤裸鼠对紫杉醇的敏感性。结论 siRNA可有效沉默A549/TXL20肺腺癌耐药细胞株LRP基因的表达,提高耐药的肺腺癌细胞对紫杉醇的敏感性。Objective To explore whether silencing lung resistance protein（LRP） may improve the sensitivity to taxol（TXL） in lung adenocarcinoma. Methods Building the taxol-resistance cell line, the graduated concentra- tions of taxol was incubated with the cells. Then the siRNA technology to silence LRP gene in A549/TXL20 was performed. MTT detected the ICs0 of taxol after A549/TXL20 transfection. Using qPCR to test the level of expres- sion of LRP mRNA. Western blot detected the protein of LRP in the cells. To build the nude mice model of trans- planted tumor by injecting A549/TXL cell into the armpit of nude mice, then detected the function of siRNA aimed at LRP to the resistance of drug in human lung adenocarcinoma cell line, A549/TXL20. Results After siRNA silencing LRP gene in A549/TXL20 cell line, the control group showed the inhibiting effect of LRP mRNAand protein expression （P 〈 0.01 ）. The sensitivity for taxol in A549/TXL20 was raised. A549/TXL20 cell line after interference failed to grow and form transplanted tumors（P 〈 0.01 ）. Conclusions siRNA may silence the LRP in A549/TXL20 which is the gene related to lung adenocarcinoma resistant drugs, and raise the sensitivity of the cell line to taxol.

关 键 词：人肺腺癌细胞株 紫杉醇 肺耐药相关蛋白 RNA干扰 

分 类 号：R734.2[医药卫生—肿瘤]