论文部分内容阅读
目的探讨Wnt/β-catenin信号通路在胚胎癌细胞增殖中的作用及其机制。方法取小鼠胚胎癌细胞系P19进行传代培养,将细胞分为正常对照组(Con组)、添加反式维甲酸组(RA组)、添加GSK-3β特异性抑制剂SB216763组(SB组)和添加SB216763+RA组(SB+RA组)四组。采用免疫荧光染色、RT-PCR和Western blot-ting法分别检测细胞中β-catenin、Oct4及C-myc的表达状况;通过BrdU标记与MTT法检测细胞的增殖。结果 加入SB216763可促进P19细胞β-catenin入核,同时促进Oct4和C-myc基因的表达,阻断RA诱导的细胞分化,并可明显促进细胞增殖。结论 激活Wnt/β-catenin信号通路可促进胚胎癌细胞P19的增殖并抑制分化,其作用机制可能是通过上调C-myc基因表达来实现的。
Objective To investigate the role of Wnt / β-catenin signaling in the proliferation of embryonic carcinoma cells and its mechanism. Methods The mouse embryonic carcinoma cell line P19 was subcultured and divided into normal control group (Con group), trans retinoic acid group (RA group) and GSK-3β specific inhibitor SB216763 group (SB group) And four groups of SB216763 + RA group (SB + RA group) were added. The expression of β-catenin, Oct4 and C-myc were detected by immunofluorescence staining, RT-PCR and Western blotting respectively. The proliferation of cells was detected by BrdU labeling and MTT assay. Results SB216763 could promote the β-catenin entry into P19 cells, promote the expression of Oct4 and C-myc genes, block the RA-induced cell differentiation and promote cell proliferation significantly. Conclusion Activation of Wnt / β-catenin signaling pathway can promote the proliferation and suppress the differentiation of P19 in embryonic carcinoma cells. Its mechanism may be through the up-regulation of C-myc gene expression.