PTD-OD-HA融合蛋白对BCR-ABL阳性细胞增殖的影响  

Effects of PTD-OD-HA fusion protein on the proliferation of BCR/ABL positive cell lines

作　　者：黄峥兰[1] 季茂胜[1] 袁颖[2] 黄世峰[1] 刘钉宾[1] 曾建明[1] 温健萍 冯文莉[1] 

机构地区：[1]重庆医科大学临床血液学教研室,临床检验诊断学教育部重点实验室,重庆400016 [2]重庆医科大学附属第一医院普外科,重庆410006 [3]Canadian Blood Services, Hamilton, ON LSS 1H8 Canada

出　　处：《解放军医学杂志》2010年第7期849-852,856,共5页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(30670901)

摘　　要：目的研究PTD-OD-HA融合蛋白对BCR-ABL阳性细胞株增殖的影响。方法将前期制备的PTD-OD-HA融合蛋白作用于两种BCR-ABL阳性细胞株BaF3-P210和K562,同时设立PBS和OD-HA融合蛋白对照组,用MTT法检测细胞的生长情况。48h后离心收集细胞,用瑞氏染色及透射电镜观察细胞形态改变,流式细胞仪(FCM)检测细胞周期变化,RT-PCR、Westernblot-ting检测c-myc、cyclinD1mRNA及蛋白表达的变化。结果 MTT检测发现,与PBS和OD-HA对照组比较,PTD-OD-HA能抑制BCR-ABL阳性细胞株的生长。瑞氏染色及透射电镜观察见PTD-OD-HA处理的细胞胞质内有大量空泡样变,出现大量的溶酶体,线粒体广泛扩张;FCM检测发现G1期细胞比例增加,S期和G2期细胞比例减少;RT-PCR及Westernblotting检测发现c-myc、cyclinD1的mRNA及蛋白水平均降低。结论 PTD-OD-HA融合蛋白可抑制BCR-ABL阳性细胞生长,使细胞形态发生改变,并使细胞阻滞在G1期,这些改变可能是通过下调c-myc、cyclinD1的表达实现的。Objective To study the effects of PTD-OD-HA fusion protein on the proliferation of BCR/ABL positive cell lines.Methods PTD-OD-HA fusion protein had been purified formerly.The BCR/ABL positive cell lines BaF3-P210 and K562 were then treated with PTD-OD-HA fusion protein,and PBS and OD-HA fusion protein treated groups were set as control at the same time.The cell growth was detected by MTT test.Forty-eight hours after treatment,cells were collected,the cell morphology was observed with Wright′s stain and transmission electron microscopy （TEM）,the cell cycles were determined by flow cytometry （FCM）,and the levels of c-myc and cyclin D1 were determined by RT-PCR and Western blotting.Results MTT test revealed that,compared to PBS and OD-HA fusion protein treated groups,PTD-OD-HA fusion protein inhibited the growth of BCR/ABL positive cells;a large number of vacuoles were found in the cytoplasm after PTD-OD-HA protein treatment,and lysosome increased significantly and mitochondria expanded extensively in BaF3-P210 cells.FCM showed that cells of G_1 phase increased,while of S phase and G_2 phase decreased.RT-PCR and Western blotting revealed that the mRNA and protein levels of c-myc and cyclin D1 decreased in BaF3-P210 and K562 cells.Conclusions PTD-OD-HA fusion protein can inhibit the growth of BCR/ABL positive cells,induce cell morphological changes,and the cell cycle may be blocked in G1 phase.These changes may be caused by down-regulation of the expression of c-myc and cyclin D1.

关 键 词：白血病 髓样 慢性 融合蛋白质类 bcr-abl 细胞增殖 

分 类 号：R730.54[医药卫生—肿瘤]