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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 )的治疗提供一种新的治疗策略
Objective To study the effect of PTD-BCR / ABL SH3 fusion protein transduction domain (PTD) and leukemia cell line (K5 6 2) on the expression of recombinant human HIV-1 transactivator protein (TAT) On the proliferation and apoptosis of K562 cells.METHODS The proliferation and apoptosis of K562 cells were detected by trypan blue exclusion method, flow cytometry, transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM) and in situ labeling of apoptotic cells NEL) were used to determine the proliferation, morphological changes, apoptosis and cell cycle parameters of PTD-BCR / ABL SH3 fusion protein in K562 cells.Results PTD- BCR / ABL SH3 fusion protein could inhibit the growth of K562 cells. The higher the concentration, the stronger the inhibitory effect. The trypan blue exclusion assay, cell morphology, electron microscopy, TUNEL in situ hybridization and FCM It was demonstrated that the PTD-BCR / ABL SH3 fusion protein induced apoptosis and necrosis of K562 cells, and the fusion protein blocked the G2 / M phase by FCM.Conclusion The PTD-BCR / ABL SH3 fusion protein enters K5 6 2 cells can cause apoptosis and necrosis, leading to cells Proliferation inhibition.This result may provide a new therapeutic strategy for the treatment of chronic myeloid leukemia (CML)