HCK基因在小鼠胚胎干细胞向心肌细胞分化过程中的表达  

Expression of HCK Gene in Cardiomyocyte Differentiation of Mouse Embryonic Stem Cells

作　　者：龚杰[1] 孙凤荣[1] 钱玲梅[1] 孔祥清[1] 盛燕辉[1] 杨荣[1] 曹克将[1] 

机构地区：[1]江苏省心血管病临床医学中心南京医科大学第一附属医院心脏科,南京210029

出　　处：《实用儿科临床杂志》2008年第1期19-21,共3页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金项目资助(30470709);江苏省自然科学基金项目资助(BK2004156)

摘　　要：目的探讨HCK基因在小鼠胚胎干细胞向心肌细胞分化过程中的表达及HCK基因在维持胚胎干细胞未分化状态中的意义。方法培养小鼠胚胎干细胞,并诱导其向心肌细胞分化。抽提其胚胎干细胞未分化及分化第2、4、6、8天的总RNA,应用反转录聚合酶链反应(RT-PCR)方法对HCK基因mRNA的表达进行半定量分析;同时抽提胚胎干细胞未分化及分化第2、4、6、8天的总蛋白,应用Western-blot方法对HCK蛋白的表达进行半定量分析。采用SPSS11.5软件进行统计学分析,P<0.05为有统计学差异。结果HCK基因mRNA在未分化的胚胎干细胞(D0)及分化的第2天(D2)均有表达,而在分化的第4天(D4)开始已不能检测到有表达,HCK基因mRNA的表达水平随小鼠胚胎干细胞的分化呈急剧下调趋势;HCK蛋白的表达与mRNA的表达相一致,在小鼠胚胎干细胞的分化进程中逐渐下调,至第4天已基本检测不出。结论HCK基因随着小鼠胚胎干细胞向心肌分化,表达呈下调的趋势,HCK基因可能在维持胚胎干细胞未分化状态中起重要作用。Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells. Methods Mouse embryonic stem cells were cul- tured, then induced to differentiate into cardiomyocytes. Total RNAs were isolated from mouse embryonic stem cells in the differentiation days ：0 day （ D0）, the second day （ D2 ）, the fourth day （ D4 ）, the sixth day （ D6 ）, the eighth day （ D8 ）, respectively. The levels of HCK mR- NAs were assessed by the method of semi - quantitive reverse transcriptase - polymcrase chain reaction（ RT - PCR）. In the meanwhile, Total proteins were also isolated from mouse embryonic stem cells in the differentiation DO, D2, D4, D6, D8, and the levels of HCK proteins were evaluated by Western - blot. Results HCK mRNAs could be detected in the mouse embryonic stem cells in DO and D2, however, they were undetectablc from D4 to D8. The expression of HCK mRNAs was rapidly down - regulated during cardiomyocyte differentiation of mouse embryonic stem cells. Expression of HCK proteins, which coincided with HCK mRNAs, down - regulated during differentiation and couldn＇t be detected in D4. Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down - regulated ; HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

关 键 词：HCK基因 干细胞 分化 小鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]