摘要
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 determine the reversal effects of sorafenib on multidrug resistance in resistant human hepatocellular carcinoma cell line BEL-7402/5-FU,and discuss its possible mechanism.METHODS：The inhibitory rate of BEL-7402/5-FU cells and the reversal effects of sorafenib on multidrug resistance were measured by MTT.Intracelluar accumulation of doxorubicin and rhodamine-123 were detected by flow cytometry（FCM）.MDR1,ABCG2 and LRP mRNA were detected by RT-PCR,while P-gp was detected by Western blot.RETHODS：Reversal folds（RF） of sorafenib（2.5 μg/mL） on fluoropyrimidine,epirubicin and paclitaxel were 2.54,4.92 and 2.23-fold,and relative reversal rates were 66.7%,96.5% and 76.8%,respectively.FCM showed that sorafenib significantly enhanced intracelluar accumulation of doxorubicin（P0.05） and rhodamine-123（P0.05） in BEL-7402/5-FU cells.RT-PCR indicated that the mRNA expressions of MDR1（P0.05） and ABCG2（P0.01）were significantly decreased,but LRP was not significantly changed by sorafenib（P0.05）.In addition,Western blot revealed that P-gp was significantly decreased by sorafenib（P0.01）.CONCLUSION：Sorafenib has the capacity of partially reversing MDR of hepatocellular carcinoma in vitro,which may be caused by down-regulating MDR1/P-gp and ABCG2,inhibiting the function of P-gp and enhancing intracelluar accumulation of chemotherapeutics,but the reversal effects may be irrelevant to LRP.
出处
《中华肿瘤防治杂志》
CAS
2010年第21期1743-1747,共5页
Chinese Journal of Cancer Prevention and Treatment
关键词
肝肿瘤
索拉非尼
多药耐药性
逆转
hepatocellular neoplasms
sorafenib
multidrug resistance
reverse
