蝎毒多肽对白血病干细胞MDR1mRNA和P-gp表达的影响  

Effect of PESV on Expression Levels of MDR1mRNA and P-gp in Leukemic Stem Cells

作　　者：杨向东[1] 史哲新[1] 姚芳[1] 闫理想 李德冠[3] 杨文华[1] 颜田赅[1] 王兴丽[1] 杨曦[1] 

机构地区：[1]天津中医药大学第一附属医院,天津300381 [2]天津中医药大学,天津300193 [3]中国医学科学院放射医学研究所,天津300192

出　　处：《中医药学报》2016年第1期36-40,共5页Acta Chinese Medicine and Pharmacology

基　　金：天津市科委课题(12JCQNJC09000);天津市教委课题(20100206)

摘　　要：目的:探讨蝎毒多肽(PESV)对多药耐药白血病干细胞MDR1 mRNA和P-gp表达的影响。方法:以K562/A02细胞株为例,免疫磁珠法分选对数生长期细胞后经CCK-8法确认其耐药性,将耐药的K562/A02干细胞接种于BABL/c裸鼠右腋下形成瘤块,再将瘤块皮下包埋建立耐药性稳定的白血病模型。成模裸鼠随机分为6组:模型对照组、ADM组、PESV组、ADM+高剂量PESV组(高剂量组)、ADM+中剂量PESV组(中剂量组)和ADM+低剂量PESV组(低剂量组)。模型对照组给予等体积0.9%氯化钠溶液腹腔注射,其余组予相应剂量ADM和(或)PESV腹腔注射,连续给药14天。第21天观察各组裸鼠移植瘤生长情况,流式细胞术检测各组瘤组织P-gp表达,RT-PCR法检测各组瘤组织MDR-1 mRNA表达水平。结果:K562/A02细胞经免疫磁珠分选前后的CD34+CD38-细胞比率和耐药率分别为31.5%、(60.33±10.68)μg/m L,92.8%、(58.33±9.72)μg/m L,分选后细胞干性显著提高,而耐药性无差异性损失。各组造模小鼠成瘤率100%,P-gp表达结果显示模型对照组89.8%、ADM组91.9%、PESV组88.4%、高剂量组53.9%、中剂量组78.0%、低剂量组78.7%,PESV+ADM下调P-gp表达,并与PESV呈现浓度依赖关系。MDR1mRNA的表达量PESV组>低剂量组>高剂量组>中剂量组>ADM组,其中PESV组、中剂量组、低剂量组高表达,P-gp表达与MDR1 mRNA水平具有一定的一致性。结论:PESV降低ADM作用K562/A02干细胞耐药性的强度,并与PESV剂量呈现正相关,其机制可能为PESV通过调控P-gp和MDR-1 mRNA的表达,增强了K562/A02干细胞对ADM的敏感度。Objective： To investigate the impact of PESV（ peptide extract from scorpion venom） on the expression level of MDR1 mRNA and P- gp in multi- drug- resistant leukemia stem cells. Methods： Take K562 / A02 cell line for example,immunomagnetic beads were used to select cells in log phase of growth,then confirm the drug resistance with CCK- 8 method. The drug- resistant K562 / A02 stem cells were inoculated to right subaxillary of BABL / c nude mice to form tumor block,then tumor blocks were subcutaneously embedded into the nude mice to establish stable drug- resistant leukemia models. Model mice were randomly divided into six groups： a model control group,an ADM group,a PESV group（ middle dose）,an ADM ＋ PESV high dose group（ high dose group）,an ADM ＋ PESV middle dose group（ middle dose group） and an ADM ＋ PESV low dose group（ low dose group）. The model control group received an equal volume of0. 9% sodium chloride solution by means of intraperitoneal injection,and other groups received the corresponding dose of ADM and（ or） PESV in the same way. All groups were continuously medicated for 14 days. On the 21 st day,the transplanted tumor growth was observed in nude mice in each group,and then detect P- gp expression level of the tumor tissues in each group by flow cytometry,then test MDR- 1mRNA expression level of the tumor tissues in each group by RT- PCR method. Results： Before and after K562 / A02 cells were selected with immunomagnetic beads,CD34 ＋CD38- cell ratio and drug- resistant dose were 31. 5%（ 60. 33 ± 10. 68） ug / ml,92. 8%（ 58. 33 ± 9. 72） ug / ml respectively. After the selection,commitment of the cells was significantly improved,while no difference of drug- resistance was shown. Tumor formation rate of model mice in each group was 100%,and P- gp expression results were respectively as follows： 89. 8% in model control group,91. 9% in ADM group,88. 4% in PESV group,53. 9% in high- dose group,78. 0% in middle dose group,78. 7% in the low- dose group. There

关 键 词：蝎毒多肽 K562/A02 P-糖蛋白 MDR-1 

分 类 号：R285.5[医药卫生—中药学]