【摘要】 目的:观察白血病原代细胞在顺铂作用下细胞周期变化和转染Chk1/2反义寡核苷酸对顺铂诱导原代细胞凋亡的影响。方法:用RT-PCR、westernblot及免疫组化检测白血病骨髓单个核细胞(BMMNC)中Chk1和Chk2的表达,用流式细胞仪检测顺铂作用下白血病BMMNC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BMMNC,用流式细胞仪检测转染Chk1/2反义寡核苷酸后顺铂作用下BMMNC细胞凋亡率。结果:在白血病BMMNC中,Chk1和Chk2mRNA水平均存阳性表达,Chk1蛋白表达强,而Chk2蛋白表达弱。10μmol/L顺铂作用下BMMNC出现S期阻滞,转染Chk1反义寡核苷酸可明显增加顺铂诱导下BMMNC凋亡率,转染Chk2反义寡核苷酸不能增加顺铂诱导下BMMNC凋亡率。结论:Chk1可作为白血病增敏治疗的有效靶点。更多还原

【Abstract】 Objective:To investigate the change of cell-cycle of leukemic bone marrow mononucleate cells (BMMNC) induced by DDP and role of antisense oligonucleotide targeting Chk1/2 on apoptosis of BMMNC induced by DDP.Method:To investigate the expression of Chk1 and Chk2 in BMMNC by RT-PCR, western blot and Immunohistochemical analysis, to observe the change of cell-cycle by flow cytometer after different interval in which BMMNC were treated by DDP. Apoptosis of BMMNC induced by DDP was investigated by flow cytometer after transfection with antisense oligonucleotide targeting Chk1/2 by lipofection.Result:The level of Chk1 and Chk2 mRNA and Chk1 protein had no significant difference between the leukemia and control, but the level of Chk2 protein was low. BMMNC arrested at S phase at 10uM of DDP. The rate of apoptosis of BMMNC induced by DDP can be increased by transfected with antisense oligonucleotide targeting Chk1, but without same result by transfected with antisense oligonucleotide targeting Chk2.Conclusion:Chk1 may be used as target of therapy of leukemia.更多还原