Ad-EGFP-MDR1转染对小鼠骨髓细胞的影响  

Influence of Ad-EGFP-MDR1 on Mice Bone Marrow Cells

作　　者：赵珍珍[1] 罗庆[1] 宿玉玺[2] 郑改焕[3] 刘伟[2] 金先庆[2] 

机构地区：[1]重庆医科大学附属儿童医院儿科研究所外科实验室,重庆400014 [2]重庆医科大学附属儿童医院普外科,重庆400014 [3]重庆医科大学附属儿童医院血液科,重庆400014

出　　处：《肿瘤防治研究》2010年第8期855-858,共4页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金资助项目(30330590)

摘　　要：目的通过研究Ad-EGFP-MDR1转染的骨髓细胞的细胞周期,凋亡相关蛋白表达和增殖分化有无改变,探讨基因修饰的骨髓细胞用于治疗的可行性。方法用重组腺病毒Ad-EGFP-MDR1感染小鼠骨髓细胞,采用体外集落培养观察转染骨髓细胞与未转染细胞的增殖能力。采用免疫印迹检测两组细胞有无凋亡相关蛋白Bax、Caspase-9、p53的表达差异。使用流式细胞仪检测两组细胞的细胞周期有无差异。通过锥虫蓝拒染率判断细胞存活情况。结果与对照组骨髓细胞比较,转染Ad-EGFP-MDR1的骨髓细胞体外集落形成差异无统计学意义。在转染的骨髓细胞和对照细胞中均未检测到凋亡相关基因Bax,Caspase-9,p53的表达,转染骨髓细胞与未转染对照骨髓细胞比较细胞周期差异无统计学意义(P>0.05)。锥虫蓝染色两组细胞的拒染率差异无统计学意义(P>0.05)。结论骨髓细胞转染重组腺病毒Ad-EGFP-MDR1后其细胞凋亡相关基因,细胞增殖分化能力及细胞活性无明显变化,有利于进一步开展基因修饰骨髓细胞体内回输和体内安全性检测。Objective To study the cell cycle,apoptosis associated genes and proliferation of mice bone marrow cells（BMCs）infected by Ad-EGFP-MDR1 and to approach the genetic modified BMCs in somatic cell therapy.Methods To assess colony formation ability of BMCs infected by adenovirus.Expression of Bax,Caspase-9 and p53 were measured by western-blot.Cell cycle was analyzed with flow cytometry.And trypan blue was used to evaluate cell viability.Results The colony formation ability of BMCs infected by Ad-EGFP-MDR1 was not abnormal compare with the control group.No expression of Bax,Caspase-9 and p53 were detected in all of the cells.Cell cycle of each group was no change（P0.05）.Cells stained with trypan blue showed no significant statistic difference between the two groups（P0.05）.Conclusion Ad-EGFP-MDR1 has no obvious toxic effect on BMCs.It is useful to research the safety of gene therapy in vivo.

关 键 词：多药耐药 腺病毒 骨髓细胞 集落培养 凋亡 

分 类 号：R730.54[医药卫生—肿瘤]