目的研究多效生长因子(Pleiotrophin,PTN)对Schlafen3(Slfn3)和Schlafen4(Slfn4)基因表达的负调控作用。方法采用RT-PCR和Northern blot法检测对照细胞(表达Ptn的Pten-/-小鼠胚胎成纤维细胞)与Ptn-/-细胞(Ptn表达沉默的Pten-/-小鼠胚胎成纤维细胞)中Slfn3/Slfn4基因的表达;分析人脑胶质瘤U87MG细胞和人乳腺癌MDA-MB-231细胞中Ptn与Slfn3/Slfn4基因表达水平的关系;在Ptn-/-细胞培养液中加入50ng/mlPTN,检测Slfn3/Slfn4基因的表达;用Ptn-/-细胞培养液作用对照细胞后,检测Slf3/Slf4基因的表达。结果 Ptn-/-细胞中Slfn3/Slfn4基因高表达;在U87MG和MDA-MB-231细胞中,Ptn与Slfn3/Slfn4基因的表达呈负相关;在Ptn-/-细胞培养液中加入PTN后,Slfn3/Slfn4基因的表达受到抑制;Ptn-/-细胞培养液对Slfn3/Slfn4基因的表达具有诱导作用。结论 PTN对Slfn3/Slfn4基因的表达具有负调控作用。
Objective To investigate the negative regulatory effect of pleiotrophin(PTN) on the expressionof Schlafen3(Slfn3) and Schlafen4(Slfn4)genes.Methods The expressions of Slfn3/Slfn4 genes in control cells(Pten-/-mouse embryo fibroblasts expressing Ptn gene)and Ptn-/-cells(Pten-/-mouse embryo fibroblasts with Ptn gene knocked down)were determined by RT-PCR and Northern blot.The relationship between the Ptn in human malignant glioma U87MG cells as well as human breast cancer MDA-MB231 cells and expression of Slfn3/Slfn4 genes were analyzed.The expression of Slfn3/Slfn4 genes in the Ptn-/-cells after 50 ng/ml of PTN was added into the medium and in the control cells cultured in the medium for Ptn-/-cells were determined.Results Slfn3/Slfn4 genes were highly expressed in Ptn-/-cells.In U87MG and MDA-MB-231 cells,Ptn was negatively related to the expression of Slfn3/Slfn4 genes.However,after PTN was added into the medium of Ptn-/-cells,the expression of Slfn3/Slfn4 genes was inhibited.Ptn-/-cell medium induced the expression of Slfn3/Slfn4 genes.Conclusion PTN showed negative regulatory effect on the expression of Slfn3/Slfn4 genes.