Effects of PTD-ABD fusion protein on proliferation and apoptosis in myeloid leukemia cells

康志杰; 闫金松; 崔嵩; 李国辉; 黄斯勇; 梁亮; 尹郸丹; 梁英民

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Effects of PTD-ABD fusion protein on proliferation and apoptosis in myeloid leukemia cells

更新时间：2026-01-09

- Effects of PTD-ABD fusion protein on proliferation and apoptosis in myeloid leukemia cells
- Journal of Army Medical University Vol. 36, Issue 5, Pages: 417-421(2014)
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  CLC： R394.3;R730.23;R733.7
- Published：2014
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康志杰,闫金松,崔嵩,李国辉,黄斯勇,梁亮,尹郸丹,梁英民.PTD-ABD融合蛋白对髓系白血病细胞的增殖和凋亡的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(05):417-421.

　Kang Zhijie,Yan Jinsong,Cui Song,Li Guohui,Huang Siyong,Liang Liang,Yin Dandan,Liang Yingmin.Effects of PTD-ABD fusion protein on proliferation and apoptosis in myeloid leukemia cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(05):417-421.
康志杰,闫金松,崔嵩,李国辉,黄斯勇,梁亮,尹郸丹,梁英民.PTD-ABD融合蛋白对髓系白血病细胞的增殖和凋亡的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(05):417-421. DOI：

　Kang Zhijie,Yan Jinsong,Cui Song,Li Guohui,Huang Siyong,Liang Liang,Yin Dandan,Liang Yingmin.Effects of PTD-ABD fusion protein on proliferation and apoptosis in myeloid leukemia cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(05):417-421. DOI：

摘要

目的研究癌基因BCR/ABL C末端肌动蛋白结合域(actin-binding domain，ABD）蛋白对白血病细胞增殖和凋亡的影响。方法构建含有ABD基因的重组表达载体，应用蛋白转导结构域（protein transduction domain，PTD）将蛋白转入白血病细胞内，应用Western blot与免疫荧光法确定蛋白是否进入白血病细胞，然后分别应用流式细胞仪及MTT法检测融合蛋白对白血病细胞凋亡及增殖的影响。结果成功构建PTD-ABD融合表达载体，获得高纯度PTD-ABD融合蛋白。同时构建PTD表达载体并获得高纯度表达蛋白。将PTD、PTD-ABD蛋白与白血病细胞K562、HL60共培养，Western blot与免疫荧光法确定PTD、PTD-ABD蛋白成功进入细胞内。与阴性对照比较，PTD-ABD蛋白能显著抑制HL60细胞的生长，并且对HL60细胞的凋亡具有明显的促进作用，差异具有统计学意义（

0.05），但对K562细胞的增殖及凋亡无影响（

0.05）；PTD蛋白对K562、HL60细胞的增殖及凋亡无影响，与阴性对照比较，差异无统计学意义（P0.05）。结论 PTD 转导的ABD蛋白抑制HL60细胞的增殖，并诱导细胞凋亡。

Abstract

Objective To study the function of BCR-ABL protein C-terminal actin-binding domain (ABD) on cell cycle and apoptosis in K562 cells and HL60 cells

and to provide the experimental basis for chronic myeloid leukemia (CML) treatment. Methods The recombinant expression vector harboring ABD was constructed

and then recombinant proteins were transferred into leukemia cells with protein transduction domain (PTD). The localization of recombinant proteins inside the leukemia cells was confirmed by Western

blotting and immunofluorescence assay. Growth curves and apoptosis of leukemia cells transfected by PTD-ABD fusion protein were determined by MTT assay and flow cytometry respectively. Results The PTD-ABD and PTD recombinant expression vectors were constructed successfully

and proteins with high purity were obtained. The growth of HL60 cells was inhibited and the apoptosis was accelerated by PTD-ABD fusion protein. The difference between experimental and control groups had statistical significance (

0.05). However

PTD-ABD fusion protein showed no difference on K562 cells (

0.05). In addition

PTD protein had no effect on the apoptosis and growth of K562 and HL60 cells in comparison with negative control (

0.05). Conclusion PTD-ABD fusion protein inhibits the growth and induces the apoptosis in HL60 cells.

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