人类凋亡相关新基因HDDC2功能的初步研究

摘要

To detect the function of human new gene HDDC2 to HEK293T cell.By searching the human reference sequencing in NCBI,human functional gene HDDC2 (NM_016063)with full length 1615bp was cloned from human multi-normal tissue cDNA li-brary by using PCR method.Hddc2 protein character was analysed by bioinformatics.CCK8 assay monitorsed the cell death.FACS as-say was used to evaluate cell death and to test PS externalization.Western blot test the function of HDDC2 to human apoptosis related gene PARP.Bioinformatics showed that HDDC2 was 1615bp length,and from 205 to 819 encoding 204 amino acids with a predicted molecular mass of 23.4 kd.This gene was located on chromosome 6q22.31,and encompassed 6 exons and 5 introns,it also showed that this gene locate in mitochondria.CCK8 assay showed that HDDC2 gene could induce HEK293T cell death,and this new gene could not only induce HEK293T cell PS externalization but also disrupt mitochondrial transmembrane potential tested by Facs assay. Western blot showed that cleaved PARP was observed in HDDC2 transfected HEK293 cell.Human new gene HDDC2 is a new gene which can induce HEK293T cell apoptosis.%利用NCBI数据库分析得到人类基因HDDC2的DNA序列，通过聚合酶链式反应克隆得到该功能基因，运用生物信息学分析该蛋白的特性，同时利用细胞生长曲线检测HDDC2促进HEK293细胞凋亡作用；使用流式细胞术检测HDDC2对HEK293细胞的作用以及HDDC2引起HEK293细胞线粒体膜的影响；运用Western blot检测HDDC2引起细胞凋亡相关基因PARP的作用。生物信息学预测HDDC2基因位于6q22．31，全长1615bp，从205到819有一个编码204个氨基酸的开放阅读框，编码一个分子量约23．4kd的蛋白。HDDC2基因含有6个外显子和5个内含子，定位于线粒体，且在人类正常组织中普遍表达，同时HDDC2基因能够明显引起HEK293细胞的程序性死亡，能够引起其磷脂酰丝氨酸的外翻和线粒体膜电位的下降，同时该基因能够引起细胞凋亡相关基因PARP的活化。人类基因HDDC2是一个可能参与到程序性细胞死亡相关的新基因，它的发现有助于发现新的药物作用靶点或者新的基因治疗药物。