P19细胞向心肌细胞分化过程中FHL1基因的表达变化  

Changes of the expression of FHL1 gene during the differentiation of P19 cells to cardiac myocytes

作　　者：郭英杰[1] 张奇军[1] 钱玲梅[1] 孔祥清[1] 曹克将[1] 

机构地区：[1]南京医科大学第一附属医院心脏科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2008年第5期601-604,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助项目(30470709);江苏省自然科学基金资助项目(BK2001043)

摘　　要：目的:观察FHL1(Four and a half LIM domains protein1)基因在P19细胞向心肌细胞诱导分化过程中表达水平的变化,探讨FHL1基因与心肌细胞分化之间的关系。方法:P19细胞经1%二甲基亚砜(DMSO)诱导悬浮培养4天形成细胞聚集体,将聚集体用生长培养基黏附培养至14天,观察细胞跳动情况,用肌钙蛋白I(cTnI)抗体行Western Blot以鉴定心肌细胞分化,并采用RT-PCR、Western Blot技术在细胞分化成熟的不同时段检测P19细胞中FHL1基因mRNA和蛋白的表达水平。结果:DMSO诱导4天后,用生长培养基黏附培养至第8天,P19细胞出现自发性节律跳动的细胞团片,Western Blot检测cTnI蛋白呈阳性。FHL1基因在P19细胞向心肌细胞分化过程中,随细胞分化成熟该基因表达水平逐渐上调,Western Blot检测结果与RT-PCR检测结果基本一致。FHL1基因在分化第0～8天表达显著增高,各时间点之间差异显著(P<0.05);第8～14天表达趋于稳定,各时间点之间差异无显著性(P>0.05)。结论:FHL1基因在P19细胞向心肌细胞分化过程中表达逐渐上调,可能参与心肌细胞的分化和发育。Objective：To investigate the changes of FHL1 （Four and a half LIM domains protein 1） gene mRNA and protein expression during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between FHL1 gene and the differentiation stage of cardiacmyocytes. Methods：P19 cells were cultured with DMSO in suspension for 4 days to form cell aggregation. Then the aggregates were cultured on gelatin-coated culture dishes without DMSO up to 14 days. The beating of cells was observed. Western Blot of Cardiac Troponin I （cTnl） were used to identify cells differentiating into cardiac cells. Total RNA and protein were extracted from P19 cells during the process of differentiation at various time points. The levels of FHL1 mRNA and protein were eval- uated by RT-PCR and Western blot,respectively. Results： After induced by DMSO for 4 days in suspension, spontaneous and rhythmically beating cells were seen at 8 day,which were cTnI-positive. When P19 cells were induced to differentiate into cardiac myocytes,both the expression level of FHL1 gene mRNA and protein were up-regulated gradually during 0-8 days of differentiation. The FHL1 gene mRNA and protein expression were obviously up-regulated. There were significant differences among the different days （P 〈 0.05）. From 8 th-14 th day,both the mRNA and protein expression of FHL1 gene were kept in higher level,and there was no significant differences among different days（P 〉 0.05）. Conclusion：Both the expression of FHL1 gene and protein were up-regulated in the process of the differentiation of P19 cells into cardiacmyocytes, which might be involved in the differentiation of cardiac rnyocytes.

关 键 词：FHL1基因 P19细胞 细胞分化 

分 类 号：R331.31[医药卫生—人体生理学]