蝎毒多肽逆转白血病干细胞多药耐药作用研究.docVIP

蝎毒多肽逆转白血病干细胞多药耐药作用研究 　　[摘要] 该实验采用BALB/c小鼠多药耐药（MDR）白血病模型，观察蝎毒多肽（PESV）对小鼠瘤体MDR白血病干细胞（LSC）P-gp（P-gp）蛋白上游信号的影响，研究PESV逆转白血病干细胞（LSC）多药耐药的效果及机制；同时通过流式细胞术，RT-PCR，Western blot，Elisa法分别检测 P-gp，MDR1 mRNA，PI3-K，NF-κB表达，并通过组织病理学方法检测小鼠肝脏、脾脏等。结果显示，正常对照组小鼠无明显变化，其余6组小鼠均出现弓背、消瘦等表现，肝脏肿大，肝内均出现炎症坏死；PESV下调小鼠瘤块中LSC细胞膜P-gp，PI3K蛋白表达下调；胞浆中MDR1 mRNA表达在PESV低剂量组显著下降，在中、高剂量组出现不同程度的升高；PESV显著降低LSC细胞核内NF-κB因子水平。PESV具有明显下调P-gp上游信号PI3K/NF-κB/MDR1的作用，在一定程度上增强了LSC对化疗药物阿霉素（ADM）的敏感度，从而?U述了PESV逆转LSC多药耐药的可能机制之一，为PESV联合抗肿瘤作用的进一步研究提供了基础。 　　[关键词] 蝎毒多肽；多药耐药；BALB/c小鼠模型；P-gp上游信号 　　[Abstract] Using the BALB/c mouse multidrug resistance model of leukemia，the effect of peptide extract from scorpion venom （PESV） to the upstream signal factors of P-gp of MDR leukemia stem cells on the mouse tumor block was observed，and the mechanism of PESV to reverse the MDR of LSC was studied. At the same time，the expression of P-gp，MDR1 mRNA and PI3-K，NF-κB were respectively detected through flow cytometry，RT-PCR，Western blot and Elisa，and the mouse liver，spleen were examined via histopathological methods. The results of the experiment were as follows： mice of the control group didn’t show any obvious changes，while mice of the other six groups all showed arched back，emaciation，liver swell，and inflammation was found in all liver tissue. The expression level of P-gp and PI3K on the LSC membrane of mouse tumor block was down-regulated；the expression of MDR1 mRNA in the cytoplasm was obviously down in the PESV low dose group，and which was inordinately up in the middle dose group and the high dose group. The expression level of NF-κB in the leukemia stem cell nucleus remarkably decreased. PESV had a outstanding role of down-regulating PI3K，NF-κb，MDR1 which were all upstream factors of P-gp，and to a certain degree enhanced the sensitivity of LSC to ADM. Therefore，this experiment explained one of the mighty mechanism of PESV to reverse MDR of LSC，and provided a foundation to further study of combinational anti-cancer effects of PESV.