摘要
为研究人肝癌多药耐药细胞株的耐药机理，应用ＢＥＬ－７４０２细胞株，通过不断提高培养液中阿霉素（Ｄｏｘｏｒｕｂｉｃｉｎ）的浓度，长期筛选培养，得到肝癌多药耐药株ＢＥＬ－７４０２／Ｄｏｘ。经ＭＴＴ法检测ＢＥＬ－７４０２对长春新碱（ＶＣＲ）等８种抗癌药的抗性提高了２７倍～１１００倍。采用流式细胞技术检测了细胞株表面ＭＤＲ１蛋白Ｐ－ｇｐ、多药耐药相关蛋白ＭＲＰ及谷脱甘肽硫转移酶ＧＳＴ－π的表达；用ＲＴ－ＰＣＲ方法检测了ＭＤＲ及ＭＲＰ基因表达水平。流式细胞分析发现，９３．５％～９７．４％的ＢＥＬ－７４０２／Ｄｏｘ细胞表面Ｐ－ｇｐ表达阳性；８４．７％～９０．２％的ＢＥＬ－７４０２／Ｄｏｘ细胞表面ＭＲＰ表达阳性；ＢＥＬ－７４０２细胞与ＢＥＬ－７４０２／Ｄｏｘ细胞ＧＳＴ－π的表达无明显变化。ＲＴ－ＰＣＲ证实此细胞株中有ＭＤＲ及ＭＲＰｍＲＮＡ的较高表达。
In order for establishing human liver cancer multidrug resistance cell strain and reserching its drug resistance mechanism, the BEL7402 cells were cultured by the method of progressively increasing the concentration of doxorubicin, and the drug resistance cell strain BEL7402/Dox was selected for a long period screening culture. The drug resistance of BLE7402 cells to vincristine(VCR) and other 8 anticancer drugs was increased 271100 times respectively by MTT assay. The MDR1 expression productmembrane efflux pglycoprotein, multidrug resistanceassociated protein(MRP) and GSTπ were examined. The MDR and MRP gene expression were also detected using RTPCR. The results showed that pgp levels of BEL7402/Dox were 93.5% to 97.4%, MRP were 84.7% to 90.2%, and the level of MDR and MRP mRNA expression were also relatively higher. No over expression of GSTπ was found.
出处
《肿瘤研究与临床》
CAS
1997年第4期222-225,共4页
Cancer Research and Clinic
关键词
多药耐药
相关蛋白
肝肿瘤
化学疗法
Multidrug resistance Multidrug resistanceassociated protein Liver cancer Chemotherapy
