喻 喆,吴 涛,张 阳.槐耳清膏体外逆转人肝癌耐药细胞BEL-7402/5-Fu多药耐药性[J].肿瘤学杂志,2013,19(6):443-447. |
槐耳清膏体外逆转人肝癌耐药细胞BEL-7402/5-Fu多药耐药性 |
The Reversal Effects of Trametes Robiniophila Murr. on Multidrug Resistance in Resistant Human Hepatocellular Carcinoma Cell Line BEL-7402/5-Fu |
投稿时间:2013-03-12 |
DOI:10.11735/j.issn.1671-170X.2013.06.B008 |
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中文关键词: 肝肿瘤 人肝癌细胞株 槐耳清膏 多药耐药性 P-糖蛋白 |
英文关键词:liver neoplasms human hepatocellular cell line Trametes robiniophila Murr. multidrug resistance P-glycoprotein |
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中文摘要: |
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。 |
英文摘要: |
Abstract:[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 IC50 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. |
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