细胞周期蛋白E2反义脱氧寡核苷酸对K562细胞增殖及凋亡的调控作用  

Effect of cyclin E2 antisense oligonucleotide on human leukemic cell line K562

作　　者：徐丽粉[1,2,3] 郭晓楠[1] 刘英芳 

机构地区：[1]河北医科大学第二医院血液内科,河北石家庄050000 [2]宁晋县医院内科,河北宁晋055550 [3]宁晋县医院心内科,河北宁晋055550

出　　处：《临床荟萃》2012年第1期39-41,45,共4页Clinical Focus

摘　　要：目的研究细胞周期蛋白E2(cyclin E2)反义脱氧寡核苷酸(ASON)对人红白血病细胞K562增殖的调控作用。方法采用反义技术合成ASON并与K562细胞共培养。用四甲基偶氮唑蓝(MTT)法检测转染ASON和脂质体lipofectamineTM2000后的细胞活力,逆转录-聚合酶链式反应(RT-PCR)方法检测转染细胞cyclin E2mRNA表达水平及流式细胞术和形态学观察检测细胞凋亡。结果 Cyclin E2特异的ASON能显著地抑制cyclin E2mRNA水平的表达(F=26.442,P<0.01);白血病细胞的生长明显受抑制(P<0.01),细胞凋亡明显增加。结果表明反义脱氧寡核苷酸能有效地抑制K562细胞的增殖,抑制K562细胞cyclin E2mRNA表达上调,并显著地诱导细胞凋亡。结论脂质体转染cyclin E2的反义寡核苷酸能够有效地抑制K562细胞cyclin E2mRNA的表达,同时对白血病细胞K562的生长有明显的抑制作用,并可诱导K562细胞凋亡。提示cyclin E2在细胞周期调控中起作用,cyclinE2基因有望作为反义技术治疗急性白血病中的一个较有意义的新靶点。Objective To investigate the effect of cyclin E2 antisense oligonucleotide（ASON） on human leukemic cell line K562.Methods Cyclin E2 ASON was used in vitro culture K562 cell study.MTT assay was used to measure the growth inhibitory effect of transfection of ASON and lipofectamineTM2000.The mRNA expression levels of cyclin E2 were examined by semi-quantitative reverse transcription polymerase chain reaction（RT-PCR）.Apopsis was detected by flow cytometry and morphology method.Results Cyclin E2 ASON specifically inhibited K562 cell mRNA expression level as well as the K562 cell proliferation.After transfection with cyclin E2 ASON,K562 cells developed apoptosis.Conclusion Cyclin E2 ASON can specifically inhibit K562 cell mRNA expression levels as well as the K562 cell proliferation.After being transfected with cyclin E2 ASON,K562 cells developed apoptosis.Cyclin E2 gene is likely to be a new target for antisense nucleotides techonology therapy of leukemia.

关 键 词：白血病 细胞周期蛋白E2 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤]