摘要
目的:探讨细胞周期素A(cyclinA)的特异反义脱氧寡核苷酸(ASON)对HL60细胞及原代白血病细胞增殖的调控作用。方法:采用反义技术将cyclinA特异的ASON与HL60细胞及原代白血病细胞共同培养,用流式细胞术及RT-PCR等方法检测cyclinA蛋白及mRNA表达水平,用苔盼蓝拒染法检测细胞生长曲线、软琼脂培养法检测CFU-HL60的集落形成情况。结果:在HL60及原代白血病细胞中,cyclinA的ASON能特异地抑制其mRNA的表达及其蛋白合成,与cyclinA的正义脱氧寡核苷酸(SON)组及对照组相比,有显著性差异(P<0.01);而且当ASON的浓度达到一定程度时,白血病细胞的增殖及其CFU-L的集落形成率均明显受抑制(P<0.01)。结论:cyclinA的ASON通过封闭cyclinA的mRNA水平,使cyclinA蛋白合成减少,能抑制白血病细胞增殖,即cyclinA是调控白血病细胞增殖的关键因子。
Objective:To investigate the function of cyclin A antisense oligodeoxynudeotide(ASON)in mediating proliferation of HL60cell and primary cell of leukemia.Methods:After cyclin A ASON was used in vitro co-culture with HL60cell and primary cell of leukemia,the protein and mRNA expression levels of cyclin A were measured by flow cytometry and RT-PCR.Growth curve of leukemia cell was determined using the trypan blue dye exclusion assay.The colony formation condition of CFU-L was detected by soft agar cultivation.Results:In cyclin A ASON group the protein and mRNA expression levels of cyclin A were significantly inhibited than those in sense oligodeoxynudeotide(SON)and control groups(p<0.01).Moreover as the ASON concentration increased,the proliferation rates of HL60cell or primary cell of leukemia and CFU-L colony unit were also significantly inhibited.Conclusions:cyclin A ASON can specifical-ly inhibite its mRNA expression levels and protein synthesis as well as the AL cell proliferations.Cyclin A is a key factor on regulation of leukemia cells proliferation.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2003年第11期772-775,共4页
Chinese Journal of Clinical Oncology
基金
河北省科委科研基金(02276104d-13)
广西卫生厅科研基金资助(编号:Z2001019)
