多胺生物合成抑制诱导HL_(60)细胞凋亡相关基因的克隆及表达

Cloning and expression of a gene associated with HL 60 cell apoptosis induced by inhibition of polyamine biosynthesis

目的克隆多胺生物合成抑制剂二氟甲基鸟氨酸（ＤＦＭＯ）诱导ＨＬ６０细胞凋亡的相关基因，分析该基因表达及诱导凋亡的活性。方法采用差示杂交法筛选目的基因克隆；以Ｎｏｒｔｈｅｒｎ杂交、形态观察、流式细胞光度法（ＦＣＭ）分析及ＤＮＡ电泳梯形图，分别证明转染细胞中该基因表达及凋亡诱导。结果成功地克隆了ＤＦＭＯ诱导ＨＬ６０细胞凋亡的相关基因ｄＦ４，并证明了ｄＦ４在转染细胞中的表达及其诱导凋亡的活性。结论提示本研究克隆的ｄＦ４基因可能是与调控ＨＬ６０细胞凋亡有关的基因。

Objective To clone the gene associated with apoptosis induced by an inhibitor of polyamine biosynthesis, α difluoromethylornithine (DFMO). Methods The differential substraction screening was used for gene cloning. The gene expression and apoptosis of transfected HL 60 cells were detected by Northern blot, morphological assay, FCM and ladder map of DNA electrophoresis, respectively. Results An apoptosis associated gene named dF4 was cloned from HL 60 cells treated with polyamine biosynthesis inhibitor. The programmed cell death was demonstrated in the HL 60 cells transfected by dF4 gene. Conclusion dF4 gene cloned in this study could be a gene regulating apoptosis of HL 60 cells.