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EGCG对人耐药口腔表皮样癌细胞株耐药逆转的实验

MDR-reversing Effect of (-)epigallocatechin-3-gallate on Human MDR Cell Lines KBV200

  • 摘要: 目的 研究EGCG对人多药耐药口腔癌细胞KBV200的细胞毒增敏作用及裸鼠移植瘤的抑瘤作用。方法 MTT法检测药物对细胞的毒性作用,流式细胞术分别检测细胞P糖蛋白的表达,HPLC检测细胞内VCR浓度,采用KB和KBV200细胞分别种植同一裸鼠左、右腋下,观察用药后体重、抑瘤率的改变。RT-PCR检测瘤组织mdr1的表达。结果 EGCG在100mg·L-1以下剂量对两株肿瘤细胞的抑制率均小于10%,EGCG与VCR联合应用可明显提高VCR的细胞毒作用;EGCG联合VCR作用后KBV200细胞内VCR浓度升高,P糖蛋白的表达下降;EGCG可增加VCR对KBV200的抑瘤作用,可降低瘤组织MDR1的表达量。结论 EGCG可增强VCR对多药耐药肿瘤细胞KBV200的细胞毒作用,机制可能与降低MDRI-mRNA、P-gp表达,提高细胞内药物浓度有关。

     

    Abstract: Objective  Experiment s were carried out to examine the potential of EGCG(epigallocatechingallate) as a multidrug resistance (MDR) reversal agents. Methods  MTT assay was used to detect cytotoxicity of EGCG and vincristine (VCR) . Int racellular concent ration of VCR was detected by high performance liquid chromatograghy( HPLC) . Flow cytomet ry was used to determine the expression of P-gp.In a BALB/ C-nu/ nu mouse model, cells of drug-sensitive KB and KBV200 (MDR) cell lines were inoculated to yield tumors in opposite flanks. EGCG and VCR were injected to the peritoneal of nude mice with carcinoma xenograf t s. MDR1 mRNA expression was observed with reverse-transcriptase PCR. Results  Survival of cells incubated with EGCG at 75 mg/ l for 72 h was over 80 %. EGCG at 8 mg/ l almost completely reversed resistance to VCR in KBV200 cells and produced a 13. 0-fold reversal of MDR. It increased int racellular concent ration of VCR in KBV200 cells while not influence that in KB cells. In KBV200 xenograf t model, neither EGCG nor VCR inhibited tumor growth. However, VCR and EGCG combined inhibited tumor growth by 72. 8 %. EGCG inhibited MDR1 expression and augmented accumulation of VCR in KBV200 cells. Conclusion  The result s suggest that EGCGis a potent MDR-reversing agent in vit ro and in vivo. The mechanism is probably associated with down-regulating the expression of MDR1 and P-gp . So that increases the concent rations of anticancer drug in tumor cells.

     

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