TATPTD-BCR/ABL SH3融合蛋白对白血病细胞系K562凋亡诱导作用

TAT PTD-BCR/ABL SH3 fusion protein induces the apoptosis of K562 leukemic cell line

目的 研究基因重组 HIV- 1反式激活蛋白 (TAT)的蛋白转导域 (PTD)与 BCR/ ABL SH3结构域融合蛋白 (PTD-BCR/ ABL SH3)导入白血病细胞株 (K5 6 2 )的细胞后对 K5 6 2细胞增殖、凋亡的影响 .方法 通过台盼蓝拒染法、流式细胞术、透射电镜 (TEM)、激光共聚焦显微镜 (CL SM)及凋亡细胞原位标记 (TU NEL)等方法 ,测定了 PTD- BCR/ ABL SH3融合蛋白作用于 K5 6 2细胞后细胞的增殖、形态学变化、细胞凋亡和细胞周期参数的改变 .结果 ≥ 5μmol· L- 1 的 PTD-BCR/ ABL SH3融合蛋白对 K5 6 2细胞生长有抑制作用 ,浓度越高 ,抑制作用越强 ;苔盼蓝拒染法、细胞形态学、电镜超微结构、TUNEL原位杂交和 FCM检测的观察证明 ,PTD- BCR/ABL SH3融合蛋白可引起 K5 6 2细胞的凋亡和坏死 ;FCM检测显示该融合蛋白可将细胞阻滞在 G2 / M期 .结论 PTD-BCR/ ABL SH3融合蛋白进入 K5 6 2细胞后可引起细胞凋亡和坏死 ,导致细胞增殖抑制 .这一结果可能为慢性粒细胞性白血病 (CML )

AIM To investigate the effect of genetic recombined HIV 1 TAT protein transduction domain (PTD) and BCR/ABL SH3 domain (PTD BCR/ABL SH3) fusion protein on the cell proliferation and apoptosis of K562 leukemia cell line after its transduction into K562 cells. METHODS The morphology, apoptosis and cell cycle of K562 cells were detected by means of trypan blue tropchrome cells, flow cytometry, transmission electron microscopy, confocal laser scanning microscopy and tunel in situ hybridization to investigate the changes of cell proliferation after PTD BCR/ABL SH3 fusion protein cultured with K562 cells. RESULTS ≥5 μmol·L -1 PTD BCR/ABL SH3 fusion protein inhibited the proliferation of K562 cells and the higher the concentration of fusion protein, the stronger the inhibitory effect was. CONCLUSION PTD BCR/ABL SH3 fusion protein could induce the apoptosis and necrosis of K562 cells and lead to the inhibitory proliferation of K562 cells. It seems to offer a new therapeutic strategy for chronic myelogenous leukemia (CML).