摘要
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 explore the role and mechanism of the Wnt/β-catenin pathway on proliferation of embryonic carcinoma cells（EC）. Methods P19 calls, a mouse EC cell line, were divided into four groups： the normal control group（con）, the RA group （medium with RA）, the SB group（medium with GSK-3β inhibitor SB216763）, and SB ＋ RA group （medium with both SB216763 and RA）. At different culture times, immunofluorescenee staining, RT-PCR and Western blotting were employed to observe expression of β-catenin, Oct4 and C-myc. BrdU labeling and MT＇F were used to measure eel/proliferation. Results The addition of SB216763 improved the nuclear location of β-catenin, high expression of Oct4 and C-myc, blocked P19 cell differentiation induced by RA, and dramatically promoted P19 cell proliferation. Conclusion Activation of the Wnt/β-catenin pathway could promote proliferation and inhibit differentiation of mouse EC cells by up-regulating C-myc expression.
出处
《山东大学学报（医学版）》
CAS
北大核心
2011年第6期53-58,共6页
Journal of Shandong University:Health Sciences
基金
国家自然科学基金资助项目(30871405
30971653)
