Triad1调节K562细胞增殖、凋亡和耐药研究

Triad1 regulates cell proliferation, apoptosis, and chemo-resistance In K562 cells

目的:研究Triad1对慢性髓细胞白血病(CML)细胞株K562增殖和凋亡的作用以及与伊马替尼(IM)耐药的关系。方法:将K562细胞进行血清饥饿培养,流式细胞仪检测细胞周期变化,蛋白免疫印迹法检测PCNA等增殖相关蛋白的表达。sh RNA干扰K562细胞后细胞周期和凋亡的变化。构建耐药细胞K562R,蛋白免疫印迹法检测K562和K562R细胞中Triad1以及凋亡相关蛋白的表达。结果:(1)血清饥饿培养后K562细胞阻滞在G0/G1期,恢复血清培养后随着K562细胞增殖,Triad1表达减少,而PCNA等蛋白表达增加。(2)干扰Triad1表达后,K562细胞增殖能力增强,G1期细胞减少。(3)K562R细胞Triad1的表达下降,凋亡相关蛋白表达降低。结论:Triad1通过调节细胞周期来调节K562细胞的增殖,干扰Triad1可促进K562细胞增殖,凋亡减少,Triad1表达下降可能与K562细胞耐药相关。

Objective： o investigate the effects of Triad1 on the proliferation and apoptosis of K562 cells in chronic myeloid leukemia（CML） and its relationship with imatinib（IM）-mediated CML cell resistance, and to explore its mechanism and significance. Methods： The K562 cells underwent serum starvation. Cell cycle progression was determined by flow cytometry. The changes in expression of PCNA in the cell proliferation was measured by Western blot analysis.K562 cells were transfected with either triad1 sh RNA or control sh RNA. Triad1 level was significantly decreased with sh RNA transfection, which was determined by Western blot. Flow cytometry was used to measure cell cycle distribution and apoptosis. K562 cells were added into imatinib（IM） according to different concentration gradients to construct imatinib（IM）resistant cell K562 R. Western blot analysis was used to detect the level of Triad1 and related protein. Results：（1）After serum deprivation in K562 cells, the cell cycle was arrested in G0/G1 phase after re-feeding. With the stimulation of serum in K562 cells, the expression of Triad1 was decreased. The expression of PCNA is increased.（2）Cell proliferation ability was promoted when expression of Triad1 was lost. Cells in G1 phase were decreased.（3）The expression of Triad1 was decreased with imatinib resistance in K562 R, and the level of apoptosis-related protein was decreased. Conclusions：Triad1 regulated the proliferation of K562 cells via the regulation of cell cycle. Knockdown of Triad1 promoted the proliferation of K562 cells. Triad1 may promote apoptosis in K562 cells. The expression of Triad1 is related to the resistance of K562 cells.