骨形成蛋白4对人牙髓细胞去分化基因表达的调控作用  

The modulatory mechanism of bone morphogenetic protein 4 on the expression of reprogramming markers in human dental pulp cells

作　　者：刘路[1] 韦曦[1] 凌均棨[1] 张芳[1] 吴莉萍[1] 

机构地区：[1]中山大学光华口腔医学院.附属口腔医院.口腔医学研究所,广州510055

出　　处：《中华口腔医学研究杂志（电子版）》2011年第5期1-5,共5页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：国家自然科学基金(81070830);广东省自然科学基金(9151008901000199);广东省医学科研基金(2009223)

摘　　要：目的研究外源性骨形成蛋白4(BMP-4)对体外连续传代培养人牙髓细胞(DPCs)生长和去分化基因表达的影响,探讨BMP-4对DPCs体外传代培养过程中细胞未分化性能的调控作用。方法重组人骨形成蛋白(rhBMP-4)诱导组和对照组体外培养4周的人牙髓组织中的细胞进行细胞计数;取P2和P7代rhBMP-4诱导组和对照组DPCs,免疫荧光染色和实时荧光定量聚合酶链反应检测Oct-4、Sox-2、c-Myc表达情况,统计学分析。结果体外培养4周的人牙髓组织经rhBMP-4诱导后,DPCs数量显著高于对照组(P<0.05)。免疫荧光示Oct-4、Sox-2、c-Myc在P2代rhBMP-4诱导组和对照组DPCs均为细胞核表达,在P7代诱导组DPCs保持细胞核表达,而在P7代对照组DPCs表现为细胞浆表达。实时荧光定量聚合酶链反应显示Oct-4、Sox-2、c-Myc在P2代诱导组和对照组DPCs表达差异无统计学意义(P>0.05),而在P7代诱导组DPCs表达显著高于对照组(P<0.05)。结论 BMP-4可促进DPCs生长,维持传代后期去分化基因Oct-4、Sox-2、c-Myc的表达。Objective The purpose of this study was to investigate the modulatory role of exogenous bone morphogenetic protein 4（BMP-4） on the growth and the expression of reprogramming markers in human dental pulp cells with long term in vitro culture.Methods Cell counting was applied to the DPCs growing our from human dental pulp tissue with 4 w in vitro culture,either stimulated with recombinant human BMP-4（rhBMP-4） or without.Immunofluorescent staining and quantitative Realtime PCR were applied to investigate the expression of Oct-4,Sox-2 and c-Myc expression on DPCs at passage 2 and 7 of rhBMP-4 induced and control group.Results The number of out growing DPCs after 4 w in vitro cultured with rhBMP-4 was statistically higher than the one from control group （P0.05）.Oct-4,Sox-2 and c-Myc showed a similar expressingpattern in DPCs from the rhBMP-4 induced and control group at passage 2,which showed their positive nucleus expression.Whereas they lost nucleus location and showed weak cytoplasm expression in DPCs at passage 7 in control group,albeit remained necleus location in DPCs at passage 7 from the rhBMP-4 induced group.The mRNA expression of Oct-4,Sox-2 and c-Myc was similar in DPCs from both rhBMP-4 induced and control group （P0.05）,whereas their mRNA expression were all sinificantly higher in DPCs from rhBMP-4 induced group than the control group （P0.05）.Conclusions BMP-4 can stimulate the growth of DPCs,and enhance the expression of Oct-4,Sox-2 and c-Myc in DPCs with long term in vitro culture,implying BMP-4 may regulate the undifferentiated capacity of DPCs with in vitro culture.

关 键 词：骨形成蛋白4 牙髓细胞 去分化基因 

分 类 号：R780.2[医药卫生—口腔医学]