视黄酸阻滞胎鼠腭间质细胞周期进程并诱导凋亡

目的探讨全反式视黄酸(atRA)对小鼠胚胎腭间质细胞(MEPM)细胞增殖和细胞周期的影响及其分子生物学作用机制。方法MEPM细胞取材于孕13d胎鼠腭组织。MTT比色法检测atRA对MEPM细胞增殖活力的影响;流式细胞术分析atRA对细胞周期分布的影响,并同时观察有无细胞凋亡现象发生;Western-blot检测atRA对细胞周期蛋白D和E表达的影响,并观察对视网膜母细胞瘤蛋白(Rb)磷酸化的影响。结果与溶剂对照组相比,在5~10μmol/L浓度范围内,atRA能够显著性抑制MEPM细胞增殖活力(P<0.05),并将细胞周期滞留在G0/G1期(P<0.05)。atRA对细胞周期蛋白D和E的表达及相应酶活性均表现为抑制效应。观察Rb的变化也显示,atRA降低Rb的磷酸化作用。结论atRA对腭器官发育敏感期腭间质细胞增殖活力及细胞周期的抑制作用可能是atRA诱导诱导唇腭裂的机制之一。

Retinoic acid induced cell cycle arrest and apoptosis in mouse embryonic palatal mesenchymal cells

Objectives To investigate the effect of all-trans retinoic acid (atRA) on proliferation activity and cell cycle distribution in mouse embryonic palatal mesenchymal (MEPM) cells and the underlying molecular mechanisms. Methods MEPM cells were prepared from palate shelves of mouse fetal on gestation day 13. Cell viability was determined by MTT assay. Cell cycle distribution and subdiploid population were analyzed by cytometry. The expression of cyclin D and E and phosphorylation of retinoblastoma protein was examined using Western-blot. Results atRA remarkably inhibited the growth of MEPM cells in a dose-dependent manner. atRA also caused an increase in the proportion of cells in G0/G1 and a decrease in the proportion of cells in S phase. atRA inhibited expression of cyclins D and E at protein level. Furthermore, atRA treatment reduced phosphorylated Rb. Conclusion These data suggested that atRA had antiproliferative activity by modulating G1/S cell cycle regulators and by inhibition of Rb phosphorylation in MEPM cells, which might account for the pathogenesis of cleft palate induced by retinoic acid.