摘要
目的:研究重组反义c-myc腺病毒(Ad-AS-c-myc)对人急性早幼粒HL-60细胞系的c-myc、增殖细胞核抗原(PCNA)表达及细胞生长抑制。方法:采用重组腺病毒Lac-Z(Ad-LacZ)及Ad-AS-c-myc联合多聚阳离子转染HL-60细胞,X-gal染色判断转染效率。采用MTT、RT-PCR及免疫细胞化学等方法进行研究。结果:多聚阳离子可提高腺病毒对HL-60细胞的转染,Ad-LacZ加入鱼精蛋白硫酸盐转染率达79.8%。Ad-AS-c-myc能抑制HL-60细胞c-myc基因在转录及蛋白水平的表达。Ad-AS-c-myc抑制HL-60细胞生长及活力(抑制率51%),细胞生长停滞后PCNA蛋白表达降低。结论:Ad-AS-c-myc对HL-60细胞具有使生长减缓、增殖能力降低作用,其生物学作用与HL-60细胞c-myc和PCNA的表达受抑有关。Ad-AS-c-myc在急性早幼粒白血病基因治疗中具有潜在的临床价值,其生物学活性与HL-60细胞c-myc和PCNA表达受抑有关。
OBJECTIVE： To study recombinant antisense c-myc adenovirus （Ad AS-c-myc） how to cause the alterations of e-myc and proliferating cell nuclear antigen expression in HL-60 cells. METHODS： The cultured HL-60 cells treated with Ad-AS-c-myc or Ad-LacZ with protamine sulfate were analysed by X-gal staining, MTT, RT PCR and immunocytochemical techniques in vitro. RESULTS： The HL-60 cells were transducted effectively by Ad-LacZ＋protamine sulfate （79. 8%）. The levels of c myc transcription and the version were strongly inhibited by Ad-AS-c-myc in the transfected HL-60 cells. Ad-AS-c myc strongly inhibited the cell growth in HL-60 cells（51%）. Ad-AS-c myc also decreased the expression of PCNA in HL-60 cells. CONCLUSIONS：Ad-AS-c myc can inhibit the growth of HL-60 cells in vitro. Ad AS-c-myc relates to the activity of PCNA and c myc gene closly and takes a key role of biologic effects in HL 60 cells. Ad AS-c-myc is a potential clinical utility in gene therapy for acute promyelocytic leukemia.
出处
《中华肿瘤防治杂志》
CAS
2008年第21期1643-1646,共4页
Chinese Journal of Cancer Prevention and Treatment
关键词
寡核苷酸类
反义
基因
MYC
腺病毒科
重组
遗传
增殖细胞核抗原
oligonucleotides, antisense
genes, myc
adenoviridae
recombination, genetic
proliferating cell nuclear antigen
