H1, a novel derivative of tetrandrine reverse P-glycoprotein-mediated multidrug resistance by inhibiting transport function and expression of P-glycoprotein
H1, a novel derivative of tetrandrine reverse P-glycoprotein-mediated multidrug resistance by inhibiting transport function and expression of P-glycoprotein
复制标题
DOI：
10.1007/s00280-010-1397-7
复制
发表时间：
2011-05
影响因子：
3
通讯作者：
Ning Wei;Hua Sun;Fengpeng Wang;Geng-tao Liu
中科院分区：
文献类型：
--
作者：
Ning Wei;Hua Sun;Fengpeng Wang;Geng-tao Liu
目的H1是粉防己碱(Tet)的新衍生物。方法采用KBv200、MCF-7/ADR及其亲本敏感细胞株KB、MCF-7进行逆转研究。用流式细胞仪检测阿霉素和罗丹明123在Pgp底物上的细胞内蓄积和外排。用Western印迹和RT-PCR方法检测Pgp的表达。用PgP-Glo™分析系统检测Pgp的ATPase活性。免疫沉淀法测定Pgp泛素化水平。结果H1显著增强KBv200和MCF-7/ADR细胞对Pgp底物的敏感性，但对亲本细胞KB和MCF-7无明显影响。H1以剂量依赖方式抑制KBv200细胞中Pgp的表达，但对MDR1的表达无影响。进一步的研究表明，H1通过促进Pgp的泛素化而促进Pgp的降解，减少Pgp蛋白的半衰期，这可能与下调MEK-ERK信号通路有关。结论H1通过抑制Pgp的转运功能和表达，是逆转Pgp介导的多药耐药的有效药物。
PurposeH1 is a novel derivative of tetrandrine (Tet). Here we investigate the ability of H1 to reverse P-glycoprotein (Pgp)-mediated multidrug resistance (MDR) and its mechanisms.MethodsKBv200, MCF-7/adr and their parental sensitive cell lines KB, MCF-7 were used for reversal study. The intracellular accumulation and efflux studies with Pgp substrates of doxorubicin and rhodamine 123 were determined by flow cytometry. The expression of Pgp was investigated by Western blot and RT-PCR analysis. ATPase activity of Pgp was performed by Pgp-Glo™assay systems. The ubiquitination level of Pgp was determined by immunoprecipitation analysis. The effect of ERK1/2 on Pgp expression in KBv200 cells were investigated by RNA interference.ResultsH1 significantly potentiated the sensitivity of Pgp substrates in KBv200 and MCF-7/adr cells, but not in parental cells KB and MCF-7. H1 inhibited Pgp expression in KBv200 cells in a dose-dependent manner, but had no effect onMDR1expression. Further studies showed that H1 prompted the degradation of Pgp and decreased Pgp protein half-life by enhancing the ubiquitination of Pgp, which may be related to downregulated MEK-ERK signal pathway. We also found H1 inhibited ATPase activity of Pgp in a dose-dependent manner.ConclusionsH1 is an effectively and potential agent in reversing Pgp-mediated MDR by inhibiting the transport function and expression of Pgp.