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目的:研究索拉非尼对人肝癌耐药细胞BEL-7402/5-FU多药耐药性逆转作用及可能机制。方法:MTT法检测索拉非尼对BEL-7402/5-FU细胞的抑制率及无细胞毒性剂量的索拉非尼对化疗药物的逆转作用;流式细胞仪检测细胞内罗丹明-123和多柔比星荧光强度;RT-PCR检测MDR1、AB-CG2和LRP等耐药基因mRNA表达情况;蛋白质印迹法检测P-gp表达情况。结果:2.5μg/mL索拉非尼对BEL-7402/5-FU细胞无细胞毒作用,对氟尿嘧啶、表柔比星和紫杉醇的逆转倍数分别为2.54、4.92和2.23倍,其相对逆转率分别为66.7%、96.5%和76.8%;流式细胞仪检测显示,BEL-7402/5-FU细胞内多柔比星(P<0.05)和罗丹明-123(P<0.05)浓度明显增加;RT-PCR显示,MDR1(P<0.05)和ABCG2(P<0.01)表达明显下降,对LRP表达无明显影响(P>0.05);蛋白质印迹法显示,P-gp表达明显下降(P<0.01)。结论:索拉非尼具有部分逆转肝癌多药耐药的作用,可能与下调MDR1/P-gp和ABCG2的表达、抑制P-gp功能以及增加细胞内化疗药物浓度有关;而此作用可能与LRP无关。
Objective: To investigate the reversal effect of sorafenib on multidrug resistance of human hepatocarcinoma cell line BEL-7402/5-FU and its possible mechanism. METHODS: The inhibitory rates of sorafenib on BEL-7402/5-FU cells and the reversal effects of sorafenib on chemotherapeutic agents were measured by MTT assay. The expressions of intracellular rhodamine-123 and The fluorescence intensity of doxorubicin was detected by RT-PCR. The mRNA expression of MDR1, AB-CG2 and LRP were detected by RT-PCR. The expression of P-gp was detected by Western blotting. Results: The cytotoxicity of 2.5μg / mL sorafenib on BEL-7402/5-FU cells was 2.54, 4.92 and 2.23 times higher than that of fluorouracil, epirubicin and paclitaxel, respectively. The relative reversal rates were (66.7%, 96.5% and 76.8% respectively). The results of flow cytometry showed that doxorubicin (P <0.05) and rhodamine-123 -PCR showed that the expression of MDR1 (P <0.05) and ABCG2 (P <0.01) decreased significantly, but had no effect on the expression of LRP (P> 0.05); Western blotting showed that the expression of P-gp was significantly decreased (P <0.01). CONCLUSION: Sorafenib can partially reverse the multidrug resistance of hepatocellular carcinoma, which may be related to down-regulating the expression of MDR1 / P-gp and ABCG2, inhibiting the function of P-gp and increasing the concentration of intracellular chemotherapeutic drugs. However, this effect may be associated with LRP Nothing to do