摘要
目的研究外源性Notch1过表达对慢性粒细胞白血病细胞株K562增殖和细胞周期的影响。方法用脂质体将携带Notch1胞内段(ICN1)的质粒转染入K562细胞,倒置相差显微镜观察转染前后K562细胞形态学变化,RT-PCR和W estern b lot检测Notch1 mRNA和蛋白的表达,MTT比色法检测细胞增殖,流式细胞仪检测细胞周期分布。结果与未转染组相比,ICN1转染组K562细胞数量减少,成团生长,直径和体积均变小;Notch1 mRNA及其蛋白表达均比未转染组增强;ICN1转染组K562细胞增殖显著受抑(P<0.01),且随时间延长而明显(P<0.05);细胞周期检测表明G1期细胞增多(P<0.05),S期细胞显著减少(P<0.01)。结论 Notch1胞内段过表达可能通过阻滞细胞于G1期而抑制K562细胞增殖。
Objective To study the effects of overexpression of exogenous Notch1 on cell proliferation and cell cycle of human chronic myeloid leukemia cell line K562.Methods Plasmid pcDNA3.1-ICN1 containing exogenous intracellular fragment of Notch1 was transfected in K562 cells with LipofectamineTM 2000.Inverted phase contrast microscope was used to observe the morphology change of K562 cells,RT-PCR and Western blot analysis were used to detect the expression of Notch1 mRNA and protein.MTT assays and flow cytometry were used to detect the proliferation and cell cycle of K562 cells respectively.Results As compared with untransfected group,the cell number of transfected K562 cells was decreased with cells aggregating.Both Notch1 mRNA and protein were up-regulated after transfection.The proliferation of transfected K562 cells were inhibited significantly（P0.01） and as we cultured more time,the cells were inhibited more significantly（P0.05）.The cell cycle assay indicated that the cells at G1 phase were increased（P0.05） and those at S phase were decreased significantly（P0.01） as compared to control group.Conclusion The overexpression of exogenous intracellular fragment of Notch1 can inhibit the proliferation of K562 cells by inhibiting the cell cycle in G1 phase.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第6期574-577,共4页
Journal of Third Military Medical University
基金
重庆市自然科学基金(CSTC2009BB5401)~~
