hhlim基因表达产物的亚细胞定位及其在细胞肥大中的意义

Abstract

hhlim是从胎儿心脏中新近分离和克隆得到的与心脏发生相关的基因 ,其表达产物作为转录因子参与多种基因的转录调控和细胞的发育与分化过程 .用细胞转染方法将外源性hhlim基因导入处于分化过程中的小鼠成肌细胞C2C12 ,发现该基因强制性表达可使C2C12细胞体积明显增大 .RT PCR和蛋白质印迹结果表明 ,hhlim促细胞肥大与诱导α 肌动蛋白 (α actin)过表达及重新启动胚胎基因BNP表达有关 .用绿色荧光蛋白 hhlim融合蛋白表达载体转染处于分化过程的C2C12细胞 ,发现转染不同时间 ,表达产物的亚细胞定位发生动态变化 ,表现为先在胞核和胞质中均有分布 ,而后定位于胞质的变化规...

hhlim was a new heart-related gene cloned from human embryonic heart whose product participated in transcriptional regulation and cell development as a kind of transcriptional factor. Over expression of hhlim gene using recombinant plasmid was sufficient to induce a greater than 1.5 fold increase in C2C12 muscle cell area compared with that untransfered with hhlim. RT-PCR and Western blot testified that transfection of hhlim into the C2C12 muscle cells could induce a-actin over expression and trigger the expression of embryonic related gene-BNP, which is related to the cell hypertrophy. hhlim-green fluorescence fusion protein in hhlim-pEGFP-C3 transfected C2C12 muscle cells was recorded with fluorescence microscopy during C2C12 muscle cell differentiation induced by horse serum. The data showed that the distribution change of green fluorescence of hhlim-GFP fusion protein from cytoplasm to nucleus was observed. Co-immunoprecipitation proved that hhlim protein exists in a manner that associates with alpha-actin in cytoplasm.