Chk1/2反义寡核苷酸对顺铂作用下K562细胞凋亡的影响

Influence of Antisense Oligonucleotide Targeting Chk1/2 on Apoptosis of K562 Cell Induced by DDP

为了观察顺铂作用下细胞周期变化规律和Chk1 2反义寡核苷酸转染K5 6 2细胞后对顺铂诱导凋亡的影响 ,采用流式细胞术检测顺铂作用下K5 6 2细胞周期变化 ;以脂质体作为载体 ,转染Chk1 2反义寡核苷酸于K5 6 2细胞 ,用Westernblot和共聚焦显微镜检测转染Chk1 2反义寡核苷酸后Chk1 2蛋白表达 ;用流式细胞术检测转染Chk1 2反义寡核苷酸后顺铂作用下细胞凋亡率。结果发现 ,10 μmol L顺铂作用下K5 6 2细胞出现S期阻滞 ,Chk1 2反义寡核苷酸对K5 6 2细胞中Chk1 2蛋白表达有明显抑制作用 ,转染Chk1 2反义寡核苷酸可明显增加顺铂诱导下K5 6 2细胞凋亡率 ,Chk1和Chk2联合转染作用优于单独转染。结论 :Chk1 2可作为白血病增敏治疗的有效靶点。

In order to investigate the change of cell-cyc le of K562 cells indu ced by cisplatin (DDP) and role of antisense oligonucleotide targeting Chk1/2 on apoptosis o f K562 cell induced by DDP, the change of cell-cycle was observed by means of f l ow cytometry after different intervals in which the K562 cell were treated by DD P . Chk1/2 protein expression was investigated by Western blot and confocal micros copy in best condition of transfection of antisense oligonucleotide targeting Ch k1/2 by lipofection. Apoptosis of K562 induced by DDP was investigated by flow c ytometry after transfection of antisense oligonucleotide targeting Chk1/2. The results showed that K562 cells were arrested at S phase at 10 μmol/L of DDP. Tr ansfection with antisense oligonucleotide targeting Chk1/2 could inhibit expression of Chk1/2 at differen t levels. The frequency of apoptosis induced by DDP was increased when transfecte d wit h antisense oligonucleotide targeting Chk1 and/or Chk2. The effect of antisense olig onucleotide targeting Chk1 and Chk2 synchronously exceeded that of antisense oli g onucleotide targeting either Chk1 or Chk2 alone. In conclusion, Chk1 and C hk2 may be regarded as targets of therapy for leukemia.