摘要
[目的]研究槐耳清膏对人肝癌耐药细胞BEL-7402/5-Fu多药耐药性逆转作用及可能机制。[方法]MTT法检测槐耳清膏对BEL-7402/5-Fu细胞的抑制率及无细胞毒性浓度的槐耳清膏对化疗药物的逆转作用;流式细胞仪检测细胞内罗丹明-123和阿霉素荧光强度;RT-PCR检测MDR1、ABCG2和LRP基因表达情况;Western blot检测P-gp表达情况。[结果]0.01mg/ml槐耳清膏对BEL-7402/5-Fu细胞无细胞毒作用,对氟尿嘧啶、表阿霉素和紫杉醇的逆转倍数分别为3.67倍、2.38倍和2.16倍,相对逆转率分别为72.8%、70.1%和74.9%;流式细胞仪检测显示BEL-7402/5-Fu细胞内阿霉素和罗丹明-123浓度明显增加;RT-PCR显示MDR1表达明显下降,对ABCG2和LRP表达无明显影响;Western blot显示P-gp表达明显下降。[结论]槐耳清膏具有部分逆转肝癌多药耐药的作用,可能与下调MDR1/P-gp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ABCG2和LRP无关。
Purpose To evaluate the reversal effect of Trametes robiniophila Murr. on multidrug resistance in drug resistant human hepatocellular carcinoma cell line BEL-7402/5-Fu,and explore its mechanism. Methods The growth inhibition of BEL-7402/5-Fu cell and the reversal effects of Trametes robiniophila Murr. were measured by MTT assay. Intracelluar accumulation of doxorubicin and rhodamine-123 were detected by flow cytometry （FCM）. The mRNA expressions of MDR1,ABCG2 and LRP were detected by RT-PCR. The expression of P-gp was detected by Western Blot.Results The IC 50 of chemotherapeutics agents in BEL-7402/5-Fu cells decreased obviously by adding non-cytotoxic concentration of Trametes robiniophila Murr. Reversal fold （RF） of Trametes robiniophila Murr. on 5-Fu,EPI and PTX were 3.67-fold,2.38-fold and 2.16-fold,respectively,and relative reversal rate （RRR%） were 72.8%,70.1% and 74.9%, respectively. FCM results showed that both Trametes robiniophila Murr. significantly enhanced intracelluar accumulation of doxorubicin and rhodamine-123 compared to the control in BEL-7402/5-Fu cells. RT-PCR results showed that the mRNA expression level of MDR1 decreased significantly after treated with Trametes robiniophila Murr. compared to the control in BEL-7402/5-Fu cells,while the mRNA expression levels of ABCG2 and LRP are not significantly changed. Western blot showed P-gp protein expression level was decreased in BEL-7402/5-Fu cells after treated with Trametes robiniophila Murr. Conclusions Trametes robiniophila Murr. can partially reverse MDR by down-regulating the mRNA and protein expression levels of MDR1/P-gp,inhibiting the function of P-gp and enhancing intracelluar accumulation of chemotherapeutic agents in human hepatocellular carcinoma in vitro. The reversal effect of Trametes robiniophila Murr. on MDR may be irrelevant to ABCG2 and LRP.
出处
《肿瘤学杂志》
CAS
2013年第6期443-447,共5页
Journal of Chinese Oncology
关键词
肝肿瘤
人肝癌细胞株
槐耳清膏
多药耐药性
P-糖蛋白
liver neoplasms
human hepatocellular cell line
Trametes robiniophila Murr.
multidrug resistance
P-glycoprotein
