摘要
目的:在全反式维A酸(all-trans retinoic acid,atRA)诱导建立腭裂小鼠模型的基础上,检测胎鼠体内胚胎腭突间充质细胞的周期分布,并检测相关周期蛋白的变化情况,为进一步阐明维A酸(retinoic acid,RA)诱导腭裂机制提供新的线索。方法:以atRA建立C57BL/6N胎鼠腭裂模型,采用流式细胞术及免疫组化检测小鼠胚胎腭突间充质细胞的周期分布,通过实时定量RT-PCR和Western印记法检测p21和pRb在胎鼠胚胎腭突间充质中的mRNA和蛋白表达水平。采用SPSS11.0软件包对数据进行单因素方差分析和t检验。结果:流式细胞术及免疫组化检测发现,在小鼠妊娠日(gestation day,GD)第10天时给予atRA,可以诱导胎鼠胚胎腭突间充质细胞在一定妊娠阶段出现G1期细胞周期阻滞,同时也使胎鼠胚胎腭突间充质内p21和pRb的表达水平出现改变。而妊娠第12天(GD12)时给予atRA则无此现象。结论:p21与pRb参与了GD10时给予atRA诱导组胎鼠胚胎腭突间充质细胞G1期阻滞的发生。
PURPOSE:To elucidate the mechanism by which all-trans retinoic acid(atRA) induces cleft palate.METHODS:The cell cycle distribution of mouse embryonic palate mesenchymal(MEPM) cells under atRA(100 mg/kg) treatment on gestation day(GD) 10 or GD 12 were measured by immunohistochemistry and flow cytometry.The p21 and phospho-Rb protein expression levels were detected by Western blot,respectively.Quantitative real-time PCR was performed for p21,pRb gene expression in each group under both conditions.The data were analyzed with SPSS 11.0 software package for one-way ANOVA and Student's t test.RESULTS:The G0/G1 arrest in MEPM cells in vivo was induced by atRA on GD 10.The protein expression levels of p21 were increased,while phospho-Rb was decreased in MEPM after atRA treatment on GD 10.These changes were not observed on the GD 12 group.Moreover,the mRNA expression levels of p21 and pRb detected by quantitative real -time PCR were almost consistent with their protein expression trends.CONCLUSIONS:The induction of G0/G1 block by atRA in MEPM cells varied with the development stage of exposure.The study demonstrated that p21 was partly required for atRA -induced cell cycle perturbations in MEPM cells.
出处
《中国口腔颌面外科杂志》
CAS
2010年第2期171-177,共7页
China Journal of Oral and Maxillofacial Surgery