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PTD-OD-HA融合蛋白对K562细胞中Bcr-Ab1的抑制作用观察
引用本文:高淼,黄峥兰,季茂胜,黄世峰,曾建明,李千音,冯文莉. PTD-OD-HA融合蛋白对K562细胞中Bcr-Ab1的抑制作用观察[J]. 解放军医学杂志, 2012, 37(10): 951-954
作者姓名:高淼  黄峥兰  季茂胜  黄世峰  曾建明  李千音  冯文莉
作者单位: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
基金项目:国家自然科学基金(30670901)~~
摘    要:目的探讨PTD-OD-HA融合蛋白转导K562细胞的动力学、定位及其与Bcr-Abl定位的关系,以及对Bcr-Abl寡聚化和Bcr-Abl酪氨酸激酶活性的影响。方法采用FITC标记PTD-OD-HA融合蛋白,观察蛋白转导K562细胞的效率与剂量和时间的关系,激光共聚焦显微镜检测融合蛋白在细胞内的定位,免疫共沉淀法检测融合蛋白与Bcr-Abl的相互作用,Western blotting检测Bcr-Abl的磷酸化水平。结果 PTD-OD-HA转导K562细胞呈剂量和时间依赖性。PTD-OD-HA定位于K562细胞胞质,与Bcr-Abl共定位,且能与Bcr-Abl蛋白相互作用,进而干扰Bcr-Abl同源寡聚化,并可降低Bcr-Abl的磷酸化水平。结论在K562细胞中,PTD-OD-HA融合蛋白能有效抑制Bcr-Abl同源寡聚化及磷酸化水平。

关 键 词:白血病,髓样,慢性  蛋白转导域  融合蛋白质类,bcr-abl

Inhibitory effect of PTD-OD-HA fusion protein on Bcr-Abl in K562 cells
GAO Miao,HUANG Zheng-lan,JI Mao-sheng,HUANG Shi-feng,ZENG Jian-ming,LI Qian-yin,FENG Wen-li. Inhibitory effect of PTD-OD-HA fusion protein on Bcr-Abl in K562 cells[J]. Medical Journal of Chinese People's Liberation Army, 2012, 37(10): 951-954
Authors:GAO Miao  HUANG Zheng-lan  JI Mao-sheng  HUANG Shi-feng  ZENG Jian-ming  LI Qian-yin  FENG Wen-li
Affiliation:Department of Clinical Hematology,Chongqing Medical University,Chongqing 400016,China
Abstract:Objective To study the transduction dynamics,location of PTD-OD-HA fusion protein and its interaction with Bcr-Abl oncoprotein in K562 cell lines,and explore the influence of PTD-OD-HA fusion protein on oligomerization and tyrosine kinase activity of Bcr-Abl.Methods PTD-OD-HA fusion protein was labeled with FITC and co-cultured with K562 cells.The transduction efficiency of labeled PTD-OD-HA at different doses and time intervals was observed under fluorescence microscope.The location of labeled PTD-OD-HA fusion protein in K562 cells was detected by confocal microscopy.The interaction of PTD-OD-HA fusion protein with Bcr-Abl oncoprotein was confirmed by coimmunoprecipitation.The phosphorylation of Bcr-Abl oncoprotein was detected by Western blotting.Results PTD-OD-HA fusion protein labeled with FITC was transduced into K562 cells in a dose-and time-dependent manner.PTD-OD-HA fusion protein was located in the cytoplasm of K562 cells and was consistent with the location of Bcr-Abl oncoprotein.The interaction of PTD-OD-HA fusion protein with Bcr-Abl oncoprotein was proved in K562 cells.This interaction could interrupt the homologous oligomerization of Bcr-Abl oncoprotein and reduce the phosphorylation of Bcr-Abl oncoprotein.Conclusion PTD-OD-HA fusion protein could be transduced into K562 cells efficiently,inhibiting the oligomerization and reducing the phosphorylation of Bcr-Abl oncoprotein.
Keywords:leukemia,myeloid,chronic  protein transduction domain  fusion proteins,bcr-abl
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