摘要
背景与目的:近几年研究发现,CRKL(CT10regulatorofkinaselike)基因编码的蛋白在慢性粒细胞白血病(chronicmyeloidleukemia,CML)发病中起重要作用。本实验研究CRKL基因反义寡核苷酸对慢性粒细胞白血病细胞系K562细胞凋亡的影响。方法:以脂质体为载体,用CRKL基因反义寡核苷酸转染K562细胞,通过活细胞计数、透射电镜、流式细胞术、DNALadder测定观察K562细胞形态学、细胞周期、基因表达等的变化。结果:在CRKL基因反义寡核苷酸作用下,K562细胞的生长明显出现抑制;流式细胞仪检测见二倍体峰前出现凋亡峰;在电镜下可见处于凋亡各期的K562细胞;提取其基因组DNA进行琼脂糖凝胶电泳,可见明显的细胞凋亡梯状条带。而对照组K562细胞未观察到上述变化。结论:CRKL基因是Ph染色体阳性CML发病的重要因素。
Background &Objective:It was recently reported that the pro teins coded by CRKL(CT10regulator of kinase like,CRK like)gene play an important role in the pathogenesis of chronic myeloid leukem ia(CML ).This study was designed to investigate the effect o f antisense oligonucleotides of CRK L gene(CRKL-ASDON)on CML K562cell lines.Methods:K562cells were transfected with CRKL-ASDON,using liposome as the vecto r.The changes of cell morphology,cell cycle,and gene expression were observed through living cell count,transmission electron microscopy,flow cytometry(FCM),DNA Ladder assay.Results:Under the action of CRKL-ASDON,grow th of K562cell was markedly inhibite d.An apoptotic peak appeared before dipl oid peak in FCM;various apoptotic stages of K562cells were observed under electron microscope;overt ladder-shaped band of cell apoptosis was seen in abstr acted DNA of the gene group by gel electrophoresis.Meanwhile,the above changes did not appear in the control.Conclusion:CRKL gene is an important factor in th e pathogenesis of Ph-positive CML.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2002年第9期934-938,共5页
Chinese Journal of Cancer
基金
国家自然科学基金(39770336)
陕西省科学基金(98SM40)
