hCG对小鼠间充质细胞(K9)转醛醇酶活性的调节  

Regulation of hCG on activity of transaldolase in K9 cell line

作　　者：袁俐[1] 张永玲[1] 袁栎[1] 张一鸣[1] 缪珩[1] 德伟[1] 

机构地区：[1]南京医科大学生物化学与分子生物学系,江苏南京210029

出　　处：《第四军医大学学报》2006年第11期1011-1013,共3页Journal of the Fourth Military Medical University

基　　金：江苏省科委国际合作项目(BZ2005043)

摘　　要：目的:研究小鼠间充质细胞(K9)中人绒毛膜促性腺激素(hCG)对转醛醇酶(TAL)活性的调节.方法:K9细胞培养,加hCG,cAMP孵化,测定TAL活性、Westernblot检测TAL蛋白表达、Northernblot检测TALmRNA表达、2DWesternblot测定TAL蛋白迁移表达.结果:①hCG促使TALmRNA表达升高.2,4,8和16hTALmRNA表达密度的扫描结果依次为:hCG组(32.7±2.3),(62.2±3.5),(27.5±2.9)和(18.1±1.1),对照组(32.3±2.7),(25.2±1.9),(31.4±3.3)和(22.5±2.6),其中4h组间差异显著(P<0.01),8及16h表达显著下降;②hCG对TAL蛋白表达无显著差异;③hCG和cAMP促使TAL活性升高,hCG组2,4,8,16和20h的TAL活性依次为(417±33),(438±47),(508±24),(576±18)和(593±29)nkat/g,均高于同期对照组(288±18),(302±20),(334±24),(393±14)和(399±10)nkat/g,组间差异显著(P<0.05);④2DWesternblot:hCG导致TAL第二亚型出现新的蛋白迁移点b6,并向酸性端迁移.结论:hCG促使小鼠K9细胞TAL的转录及活性升高,但hCG对TAL活性调节与转录水平无关,提示TAL活性调节发生在翻译后修饰,可能是通过第二信使cAMP促进了TAL磷酸化过程.AIM： To study the regulation of human chorionic gonadotropin （hCG） on the activity of transaldolase （TAL） in K9 cell line. METHODS： In this research, K9 cells were cultivated in 3 groups, the 2 of which were incubated with hCG and cAMP respectively as the experimental groups and the other 1 without any factors as the control one. The activity of TAL was measured by Lachaise and the quantity of TAL proteins was detected by Western blot, the expression of TAL mRNA by Northern blot and the migration expression of the TAL protein by 2D-Western blot. RESULTS： （ 1 ） The expression of TAL mRNA was obviously enhanced by hCG, which was manifested by the fact that there was an increase of the TAL mRNA expression by 2 h （32.7 ± 2.3） and a significant rising by4 h （62.2 ±3.5） （P〈0.05）, while by 8h and 16h remarkable drops appeared （27.5 ± 2.9, 18.1 ± 1. 1 ） respectively. In control group, the expression of TALmRNA was （32.7±2.7）, （25.2±1.9）, （31.4±3.3） and （22.5 ± 2.6） by 2, 4, 8 and 16 h respectively. （2） the expression level of TAL protein was not significantly different between hCG and control groups; （3） Both hCG and cAMP stimulated the activity of TAL, In the hCG group the activity of the TAL began to rise by 1 h of the experiment, and significantly increased by 2, 4, 8, 16 and 20 h during the process [P〈0.05 vs control group （nkat/g） ; （417 ±33） vs （288 ± 18）, （438 ± 47） vs （302 ±20）,（508 ±24） vs （334 ±24）,（576 ± 18） vs （393 ±14）,（593 ±29） vs （399 ± 10 ）, respectively]; （4） 2D- Western blot indicated that hCG led to the appearance of a new protein spot b6. CONCLUSION： hCG increases both the transcription and the activity of TAL in K9 cells, while it was not related between the activity regulation and the transcription level of TAL, which suggested that the activity regulation of TAL takes place during the post -translation modification of their proteins, and that hCG may have enhanced the ph

关 键 词：转醛醇酶 小鼠睾丸间充质细胞 人绒毛膜促性腺激素 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学] R322.6[医药卫生—基础医学]