Prdm14对C3H10T1/2细胞增殖及干性的影响

Effects of Prdm14 on proliferation and stemness of C3H10T1/2 cells

目的探讨PR结构域蛋白14(Prdm14)对C3H10T1/2细胞增殖及干性的影响。方法该实验分为空白对照组(正常C3H10T1/2细胞)、阴性对照组(感染慢病毒空载体的C3H10T1/2细胞)以及实验组(感染Prdm14慢病毒的C3H10T1/2细胞),分别采用实时荧光定量PCR(RT-qPCR)、蛋白质印迹法(WB)检测Prdm14 mRNA及蛋白水平。采用CCK-8及细胞克隆形成实验检测C3H10T1/2细胞增殖情况;通过碱性磷酸酶(ALP)染色及活性测定,观察ALP活性情况;采用WB检测干性因子Sox2、Nanog、Oct4表达。结果与阴性对照组相比,实验组Prdm14 mRNA及蛋白水平均明显升高(P<0.05),ALP活性及细胞增殖能力增强(P<0.05),干性因子Sox2、Nanog蛋白水平均明显升高(P<0.05),Oct4蛋白水平差异无统计学意义(P>0.05)。结论Prdm14可促进C3H10T1/2细胞增殖及干性因子表达。

Objective To investigate the effects of PR domain zinc finger protein 14(Prdm14)on the proliferation and stemness of C3H10T1/2 cells.Methods In the experiment,blank control group(normal C3H10T1/2 cells),negative control group(C3H10T1/2 cells infected with lentivirus empty vector)and experimental group(C3H10T1/2 cells infected with Prdm14 lentivirus)were set up,and the mRNA and protein levels of Prdm14 were detected by real time fluorescence quantitative PCR(RT-qPCR)and Western blotting(WB).The proliferation of C3H10T1/2 cells was tested by CCK8 and cell clonal formation assay.The alkaline phosphatase(ALP)staining and activity determination were performed to observe the ALP activity.The expression of Sox2,Nanog and Oct4 were detected by WB.Results Compared with the negative control group,the levels of Prdm14 mRNA and protein in the experimental group increased significantly(P<0.05),the ALP activity and cell proliferation ability increased(P<0.05),the levels of dry factor Sox2 and Nanog protein increased significantly(P<0.05),and the OCT4 protein level had no statistical significant different(P>0.05).Conclusion Prdm14 can promote the proliferation and stem factor expression of C3H10T1/2 cells.