分化抑制因子Id4对K562细胞株凋亡的影响

Effects of Id4 on Apoptosis of K562 Cell

目的探讨Id4基因的反义核酸对人红白血病K562细胞凋亡的影响。方法采用脂质体介导的反义寡核苷酸转染DNA重组技术,将人反义Id4基因插入到真核表达载体pXJ41-neo上,重组质粒和空载体分别转染K562细胞,加入凋亡诱导剂三尖杉酯碱(HT)后采用流式细胞术、TUNEL和免疫印迹(Western blot)技术检测细胞的凋亡率以及相关蛋白Bcl-xl、c-Myc、p27的表达变化。结果获得了pAId4(反义基因重组质粒)和pK562(对照)克隆细胞株;加入诱导剂HT后,反义寡核苷酸组的凋亡率明显增高到43.5%;检测K562细胞凋亡过程中,Bcl-xl蛋白在pAId4细胞株中表达减低。结论反义Id4基因的转入对K562细胞起到诱导凋亡作用,并推测Id4在K562细胞中可能通过影响Bcl-xl的表达量,进一步诱导细胞凋亡。

Objective To study the effects of Id4 on cell apoptosis of humen erythroleukemia cell line K562. Methods The antisense Id4 gene was inserted into eukaryotic expression vector pXJ41-neo, the recombinant vector was transfected into K562 cells. The percentage of apoptotic cells induced by HT was analysed by flow cytometry, UNEL. Western blot for detection of the expression of Bcl-xl, c-Myc, p27. Results The cell apoptosis was most significant in pAid4 cells induced by HT,The expression of Bcl-xl was found down-regulated in pAid4 cells in comparison with the control. Conclusion The antisense Id4 gene was found to be a strong inducer of apoptosis in K562 cells. In addition, Id4 may be an upstream regulator expression of Bcl-xl, leading to apoptosis.