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脂质体转染细胞周期素B1反义脱氧寡核苷酸对HL60细胞增殖调控的影响
引用本文:贾晋松,徐世荣,贾存荣,马劼,马卫东,姚银荣,王毅,石翠英. 脂质体转染细胞周期素B1反义脱氧寡核苷酸对HL60细胞增殖调控的影响[J]. 中国病理生理杂志, 2004, 20(2): 228-233
作者姓名:贾晋松  徐世荣  贾存荣  马劼  马卫东  姚银荣  王毅  石翠英
作者单位:1. 河北医科大学第二医院血液内科, 河北 石家庄 050000;
2. 河北省血液中心, 河北 石家庄 050000
基金项目:河北省科委基金资助项目 (No .0 2 2 76 10 4d - 13)
摘    要:目的:探讨脂质体转染细胞周期素B1(cyclinB1)反义脱氧寡核苷酸(ASON)对HL60细胞增殖调控的作用。方法:用针对cyclinB1mRNA5’端编码区起始密码子(ATG/AUG)的ASON,通过脂质体导入HL60细胞共培养后,用流式细胞术(FCM)和RT-PCR分别检测cyclinB1蛋白和mRNA的表达水平,电镜和原位细胞凋亡检测法(POD)、FCM及DNA凝胶电泳法检测细胞凋亡。结果:CyclinB1ASON组与SON及空白对照组相比,ASON能特异地抑制cyclinB1蛋白及mRNA水平的表达,当ASON的浓度达到一定程度时,HL60细胞的增殖及集落形成率均明显受抑制,出现细胞凋亡,并且此作用随ASON浓度的升高而增强。结论:CyclinB1的特异ASON能封闭其蛋白及mRNA的表达水平,可剂量依赖性地抑制白血病细胞增殖,诱导细胞凋亡。

关 键 词:细胞周期蛋白B1  寡核苷酸类  反义  脂质体  白血病  HL细胞  细胞凋亡  
文章编号:1000-4718(2004)02-0228-06
收稿时间:2003-05-19
修稿时间:2003-08-11

Effect of cyclin B1 antisense oligodeoxynucleotides with liposomal transfection on proliferation of HL60 cells
JIA Jin-song,XU Shi-rong,JIA Cun-rong,MA Jie,MA Wei-dong,YAO Yin-rong,WANG Yi,SHI Cui-ying. Effect of cyclin B1 antisense oligodeoxynucleotides with liposomal transfection on proliferation of HL60 cells[J]. Chinese Journal of Pathophysiology, 2004, 20(2): 228-233
Authors:JIA Jin-song  XU Shi-rong  JIA Cun-rong  MA Jie  MA Wei-dong  YAO Yin-rong  WANG Yi  SHI Cui-ying
Affiliation:1. Department of Hematology, the Second Hospital, Hebei Medical University, Shijiazhuang 050000, China;
2. the Blood Center of Hebei Province, Shijiazhuang 050000, China
Abstract:AIM: To investigate the effect of cyclin B1 antisense oligodeoxynucleotide (ASON) with liposomal transfection on proliferation of HL60 cells. METHODS: After cyclin B1 ASON with liposomal transfection was used in vitro co-culture with HL60 cells,the protein and mRNA expression levels of cyclin B1 were measured by flow cytometry and RT-PCR. The cell apoptosis was detected by electron microscopy, in situ cell apoptosis detection kit (POD),DNA agarose gel electrophoresis and flow cytometry. RESULTS: In cyclin B1 ASON group the protein and mRNA expression levels of cyclin B1 were significantly inhibited than those in sense oligodeoxynucleotide (SON) group and blank group. Morever,when the ASON concentration increased,the proliferation ratio of HL60 cells and CFU-HL60 clony unit were also significantly inhibited. The apoptosis of HL60 cells was also observed. CONCLUSION: Cyclin B1 ASON specifically inhibited its protein and mRNA expression levels as well as the HL60 cell proliferations and induced leukemia cell apoptosis. It's effect depended upon the concentration of ASON.
Keywords:Cyclin B1  Oligonucleotides  antisense  Liposomes  Leukemia  HL cells  Apoptosis
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