外源MDR-1基因对小鼠肺腺癌细胞耐药性产生的影响及机理研究.docVIP

外源MDR-1基因对小鼠肺腺癌细胞耐药性产生的影响及机理研究 中国肺癌杂志 2000年第1期第3卷 肺癌分子生物学研究 作者：沙慧芳　董强刚　苏建中　冯久贤　包国良 单位：沙慧芳　董强刚　苏建中　冯久贤　包国良(上海胸科医院胸部肿瘤研究所　200030) 关键词：转基因；Lewis肺癌细胞；多药耐药 　　【摘要】目的　了解MDR-1基因表达与肺癌细胞耐药性的关系。方法　用FUGENTM6转染试剂将MDR-1基因导入小鼠Lewis肺腺癌细胞株(3LL)，建立了耐药细胞株(3LL-MDR-1)。用MTT方法测定细胞耐药性并观察异搏定对耐药的逆转效应，MDR-1基因表达产物P-gp采用免疫组化方法观察，应用流式细胞仪分析其对示踪剂罗丹明123的摄取与排泄。结果　建立了一株能稳定表达P-gp和对长春生物碱类、鬼臼毒素类具有显著耐药性的细胞株。免疫组化证实耐药细胞高度表达P-gp糖蛋白。异搏定能部分逆转该细胞株的耐药性，且效应随着剂量递增，流式细胞仪测定显示耐药细胞对示踪剂的排泄作用显著高于亲代细胞。结论　通过基因转染实验，证明MDR-1基因表达与肺癌细胞多药耐药有关。 　　An establishment of murine drug-resistant cell line transfected with human MDR-1 gene SHA Huifang,DONG Qianggang,SU jianzhong,FENG Jiuxian,BAO Guoliang(Shanghai Institute of Thoracic Tumors,Shanghai Chest Hospital,Shanghai200030,P.R.China) 　　【Abstract】Objective　To investigate the relationship between MDR-1 gene expression and drug resistance in lung cancer cells.Methods　The multidrug resistant cell line (3LL-MDR-1) was established by transfecting human MDR-1 gene expressing vector into a murine lewis lung cancer cell line (3LL).Drug resistance and the effect of valapamil(VLP) on the resistance were evaluated by MTT assay.The expression of MDR-1 gene product,P-gp,was measured immunohistochemically.The influx and efflux of rhodamine 123 was tested with flow cytometry.Results　The MDR cell line expressing P-gp was established,which exhibited a typical drug resistance to vincristine (VCR),vindesin (VDS),etoposide (VP-16),mitomycin (MMC),taxol and navelbine (NOR).Valapamil was able to reverse the resistance of the gene-transfected cells and the reversion was dose-dependent.Flow cytometric analysis showed that the resistant cells had much higher ability to export rhodamine 123 than the parent cells did.The expression of P-gp protein was visualized immunohistochemically.Conclusion　The results indicate that drug resistance in lung cancer cells is closely related to the MDR-1 gene expression. 　　【Key words】Transfection gene　　Lewis lung cancer cells　　Multidrug resistance 　　肿瘤细胞多药耐药(MDR)的产生是导致化疗失败