同源盒基因Nkx2-5促进5-氮胞苷诱导的单层P19细胞表达心肌标志物  

Nkx2-5 gene promotes expression of cardiac markers in 5-azacytidine-induced monolayer P19 cells

作　　者：张金平[1] 王慧娟[1] 邵素霞[1] 尹青[1] 史玉兰[2] 赵春芳[1] 张雷[1] 

机构地区：[1]河北医科大学组织学与胚胎学教研室,石家庄050017 [2]白求恩军医学院生理学教研室,石家庄050081

出　　处：《第二军医大学学报》2009年第12期1372-1375,共4页Academic Journal of Second Military Medical University

基　　金：河北省科学技术研究与发展计划(07275555)~~

摘　　要：目的:探讨同源盒基因Nkx2-5基因在5-氮胞苷(5-aza)诱导单层生长的P19细胞向心肌细胞分化中的作用。方法:以稳定表达Nkx2-5的P19细胞为实验组,P19细胞为对照组。两组细胞均单层培养并以5-aza(1μmol/L)诱导,倒置显微镜下观察细胞的生长状态;在诱导后4、8、12、16 d,以RT-PCR检测转录因子GATA-4、α-肌球蛋白重链(α-MHC)和心房利钠多肽(ANP)基因的表达。结果:对照组经5-aza诱导后ANP没有表达;GATA-4在8、12、16 d有表达;α-MHC在12、16 d有表达。实验组5-aza诱导后,GATA-4在诱导4、8、12、16 d有表达;α-MHC和ANP都是在5-aza诱导后的8、12和16 d表达。两组结果比较显示实验组GATA-4和α-MHC基因的表达时间提前;两组除α-MHC在12 d的表达无差异外,其他取材时间点实验组两个基因的表达量与对照组相比均增高,差异有统计学意义(P<0.05,P<0.01)。结论:Nkx2-5促进5-aza诱导的单层生长的P19细胞向心肌分化。Objective：To study the role of Nkx2-5 gene in 5-azacytidine-induced differentiation of monolayer P19 cells into myocardial cells.Methods：The experiment was divided into two groups：an experimental group and a control group.The cells in the experimental group were P19 cells stably expressing Nkx2-5 gene,and cells in the control group were P19 cells.Under the monolayer-culture condition,the cells of two groups were induced by 5-azacytidine（1 μmol/L）.The growth of cells were observed by inverted microscope.On the 4^th day,8^th day,12^th day and 16^th day after induction,RT-PCR was used to detect the expression of GATA-4,α-MHC and ANP gene.Results：In control group,there was no ANP expression after induction;GATA-4 expression was seen on the 8^th day,12^th day,and 16^th day after induction;and α-MHC expression was found on the 12^th day and 16^th day.In experimental group,the expression of GATA-4 was detected on the 4^th day,8^th day,12^th day and 16^th day after induction;Alpha-MHC and ANP expression was noticed on the 8th day,12th day and 16th day after induction.RT-PCR results showed that the expression of GATA-4 and α-MHC in the experimental group was earlier than that in the control group.And at all time points of observation,the expression of GATA-4 and α-MHC in the experimental group was significantly increased compared with that in the control group（P〈0.05,P〈0.01）,except for α-MHC expression on the 12^th day.Conclusion：Nkx2-5 gene can promote 5-azacytidine-induced differentiation of monolayer P19 cells into myocardial cells.

关 键 词：NKX2-5 5-氮胞苷 P19细胞 心肌分化 

分 类 号：R321.51[医药卫生—人体解剖和组织胚胎学]