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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
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*
0
(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)
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 (1 μmol/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。
关键词:  Nkx2-5  5-氮胞苷  P19细胞  心肌分化
DOI:10.3724/SP.J.1008.2009.01372
投稿时间:2008-12-29修订日期:2009-11-24
基金项目:河北省科学技术研究与发展计划(07275555).
Nkx2-5 gene promotes expression of cardiac markers in 5-azacytidine-induced monolayer P19 cells
ZHANG Jin-ping1,WANG Hui-juan1,SHAO Su-xia1,YIN Qing1,SHI Yu-lan2,ZHAO Chun-fang1,ZHANG Lei1*
(1.Department of Histology and Embryology, Hebei Medical University, Shijiazhuang 050017, China;2.Department of Physiology, Bethune Military Medical College, Shijiazhuang 050081)
Abstract:
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 4th day, 8th day, 12th day and 16th 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 8th day, 12th day, and 16th day after induction; and α-MHC expression was found on the 12th day and 16th day.In experimental group, the expression of GATA-4 was detected on the 4th day, 8th day, 12th day and 16th 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 12th day.Conclusion:Nkx2-5 gene can promote 5-azacytidine-induced differentiation of monolayer P19 cells into myocardial cells.
Key words:  Nkx2-5  5-azacytidine  P19 cell  myocardial differentiation