文档介绍：CRKL基因反义寡核苷酸对K562细胞增殖的影响
作者:王莎,孙秉中,黄高升日,于文强,柴玉波
【关键词】反义寡核苷酸
关键词: 反义寡核苷酸;CRKL基因;慢性粒细胞性白血病
摘要:目的通过研究CRKL基因反义寡核苷酸对K562细胞增殖活性的影响,探讨CRKL基因在Ph+ 慢性粒细胞性白血病(CML)发病中的作用. 方法脂质体为载体,CRKL基因反义寡核苷酸转染K562细胞,通过活细胞记数、H3 -TdR掺入、RT-PCR等方法观察K562细胞的增殖活性及基因表达的变化. 结果 CRKL反义寡核苷酸对K562细胞的增殖有抑制作用. 结论 CRKL基因在Ph+ CML的发病中可能有重要作用,可作为CML治疗的新靶点.
Keywords:antisense-oligonucleutide;CRKL gene;chronic mylocytic leukemia
Abstract:AIM To investigate the effects of CRKL anti-sense-oligonucleutides(ASODN)on cell growth and prolifer-ation in K562cell line and thus assess CRKL gene in the pathogenesis of Ph+ K562cells were cul-tivated with CRKL oligos which were mediated by cationin li- cell viability,
proliferation,and gene expres-sion were examined by H 3 -TdR doping and RT-┐SULTS CRKL ASODN could inhibit the viability and prolif-eration of CRKL gene may play a key role in the pathogenisis in Ph+ CML.
0 引言
性粒细胞性白血病(CML)(Ph+ CML),有其形成的bcr-abl融合基因,编码的P210 bcr-abl 融合蛋白具有异常增高的酪氨酸激酶活性,通过异常激活RAS、JAK/STAT、JUN等激酶,引起调控细胞生长、发育的多条信号转导途径异常,从而使细胞发生恶性转化[1] .CRKL系bcr-abl通路中一个重要接头蛋白,为bcr-+ 的K562细胞,观察对其增殖活性的影响,以探讨CRKL基因在CML发病中的作用.
1 材料和方法
反义寡核苷酸
针对翻译起始位点的反义寡核苷酸[2] ASODN:5′-GTC T GGC GGA GGA-3
’(18bp);无意义寡核苷酸NASODN:5′-GAA GGG CTT CTG CGTC-3′(16bp).上述寡聚物均为全硫代修饰,由上海S