摘要
QKI是信号转导与RNA激活家族(STAR)成员之一,是进化保守的调控蛋白。其KH结构域能与RNA上的quaking反应元件(QRE)结合,参与调控RNA的运输、翻译、剪切、稳定等过程,从而影响细胞增殖、分化、凋亡等。本试验探究QKI是否对猪睾丸成纤维细胞(ST细胞)凋亡存在影响。构建pEF1α-QKI过表达载体,采用qRTPCR、Western blot方法检测QKI及目的基因BCLAF1的mRNA和蛋白的表达量,用流式细胞术检测过表达QKI对ST细胞凋亡的影响。结果显示:转染pEF1α-QKI质粒在ST细胞中可显著提高QKI mRNA的表达量(P<0.01),并且与对照组相比能显著降低BCLAF1mRNA和蛋白的表达量(P<0.05);流式细胞术检测细胞凋亡率证明过表达QKI具有抑制ST细胞凋亡的作用(P<0.05)。结果表明:在ST细胞中过表达QKI能够抑制BCLAF1基因的表达,并且降低ST细胞凋亡率。
QKI is the evolutionary conserved regulatory protein belonging to a member of signal transduction and RNA activation family （STAR）. Its KH domain can bind to the quaking response element （QRE） of RNA,and QKI involved in the process of RNA transport,translation, cleavage and stabilization,thus affecting cell proliferation,differentiation and apoptosis. The purpose of this study was to investigate whether QKI has an effect on the apoptosis of porcine testicular fibro- blasts （ST cells）. We constructed pEFla-QKI overexpression vector and detected the mRNA and protein expression of QKI and BCLAF1 by qRT-PCR and Western blot. Finally, flow cytometry was used to detect the effect of QKI overexpressing on apoptosis in ST cells. The results showed that transfection of pEFlorQKI plasmid significantly increased the expression of QKI mRNA in ST cells （P〈0.01）,and significantly decreased the expression of BCLAF1 mRNA and protein （P〈0.05） ,compared with the control group. In addition,overexpression of QKI can inhibit apop- tosis of ST cells. Overexpression of QKI can inhibit the expression of BCLAF1 gene and decrease the rate of apoptosis in ST cells.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2017年第3期505-508,共4页
Chinese Journal of Veterinary Science
基金
吉林省科技发展计划资助项目(20140204067NY)
