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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 to study the drug resistance mechanism of human hepatocellular carcinoma multidrug resistant cell line, BEL-7402 cell line was used, and by continuously increasing the concentration of doxorubicin in the culture medium, long-term screening culture was conducted to obtain a multidrug resistant hepatocellular carcinoma BEL- 7402/Dox. The resistance of BEL-7402 to 8 kinds of anticancer drugs such as vincristine (VCR) was increased 27- to 100-fold by MTT assay. Flow cytometry was used to detect the expression of MDR1 protein P-gp, multidrug resistance-associated protein MRP and glutathione S-transferase GST-π on the cell surface; the expression levels of MDR and MRP genes were detected by RT-PCR. . Flow cytometry analysis showed that the expression of P-gp on the surface of BEL-7402/Dox cells was positive from 93.5% to 97.4%, and the expression of MRP was positive on 84.7% to 90.2% of BEL-7402/Dox cells. There was no significant change in the expression of GST-π in BEL-7402 cells and BEL-7402/Dox cells. RT-PCR confirmed the high expression of MDR and MRP mRNA in this cell line.