肝细胞癌SMC7721体内外多药耐药模型的建立及其耐药机制

目的 建立肝细胞癌SMC7721体内外多药耐药细胞株并探讨其耐药机制.方法 通过阿霉素浓度递增筛选出SMC7721/ADM肝癌多药耐药细胞株并建立裸鼠移植瘤模型;噻唑蓝(MTr)检测各组移植瘤对化疗药物的耐药性;逆转录-聚合酶链反应(RT-PCR)和Western blot检测MDR1和BCRP在各组移植瘤中的表达.结果 SMC7721/ADM对阿霉素,丝裂霉素,5-Fu的耐药性均较亲本细胞明显降低(P<0.01);其移植瘤对3种化疗药物的IC50.均明显低于亲本细胞(P<0.05);SMC7721/ADM细胞中MDR1 mRNA表达是亲本细胞的40.13倍(P<0.01);BCRP mRNA表达与亲本细胞差异无统计学意义(P>0.05).其移植瘤中BCRP mRNA表达是亲本细胞移植瘤的3.69倍(P<0.01);而MDR1 mRNA表达与亲本细胞差异无统计学意义(P>0.05);SMC7721/ADM细胞株中两种蛋白表达显著高于亲本细胞株(P<0.01);SMC7721/ADM移植瘤中BCRP蛋白表达显著高于亲本移植瘤(P<0.01),而两者中MDR1表达差异无统计学意义(JP>0.05).结论 MDR1和BCRP在SMC7721/ADM多药耐药的不同阶段起作用并介导肝细胞癌对不同药物的耐药性.

Establishment and the possible multidrug-resistance mechanism of Hepatoma cell SMC7721 MDR model:in vitro and in vivo studies

Objective To study the role of MDR1 and BCRP in the multi-drug resistance (MDR) of hepatocellular carcinoma (HCC) in vivo and in vitro. Methods SMC7721 cell line resistant to adriamycin was induced by exposing SMC7721 cell line to gradually increased concentration of adriamycin and then transplanted in the Hypo of athymic mouse. The MDR index to chemotherapeuties of different groups was measured by the method of MTT in vivo an in vitro. The expression of BCRP and MDR1 in cell lines and their transplantation tumors was detected by RT-PCR and Western blot. Results The IC50 of SMC7721/ADM to ADM, MMC and 5-Fu were higher than that of SMC7721 obviously in vitro and in vivo(P<0.01). The expression of MDR1 mRNA in SMC7721/ADM cell lines was 40. 13 times of that in SMC7721 cells (P<0.01). The expression of BCRP mRNA in SMC7721/ADM transplantation tumor was 3.69 times of that in SMC7721 transplantation tumor (P< 0.01). The expression of MDR1 and BCRP proteins in SMC7721/ADM cell line was sinificantly higher than in SMC7721 cell line (P <0. 01). The expression of BCRP protein in SMC7721/ADM transplantation tumor was sinificantly higher than in SMC7721 transplantation tumor (P<0.01). Conclusion MDRI and BCRP play important roles in the different stages during the HCC MDR,and they are responsible for SMC'7721/ADM' s resistance to different anticancer agents.