胶质瘤细胞中上皮剪接调控蛋白1(ESRP1)对人髓细胞白血病基因(MCL1)剪接的调控  

The myeloid cell leukemia-1 mRNA splicing mediated by epithelial splicing regulatory protein 1( ESRP1) in glioma U251 cell lines

作　　者：宋大勇[1] 李志强[1] 权哲[1] 赵军[1] 许大远[1] 张宁[1] 

机构地区：[1]上海交通大学附属第六人民医院南院,上海市奉贤区中心医院神经外科,上海201499

出　　处：《细胞与分子免疫学杂志》2015年第11期1463-1467,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：上海市奉贤区科委课题(奉科20131401)

摘　　要：目的研究胶质瘤细胞系U251中上皮剪接调控蛋白1(ESRP1)对人髓细胞白血病基因1(MCL1)转录本剪接的调控作用。方法对胶质瘤患者数据库中MCL1的测序结果进行数据挖掘,寻找关键位点。构建pc DNA-ESRP1外源表达载体并在U251细胞中进行过表达,通过反转录PCR和Western blot法检测MCL1不同异构体的表达情况。结果 ESRP1在U251细胞中的蛋白表达水平较正常胶质细胞低,MCL1异构体1在U251细胞中的表达高于正常胶质细胞,而异构体2和异构体3的表达则低于正常胶质细胞;在U251细胞中过表达ESRP1以后,发现异构体1表达较未转染组下降,而异构体3表达则显著升高,异构体2表达无明显变化。此外,第801G、802A缺失的MCL1无法被ESRP1正确剪切,异构体1和异构体3的表达与ESRP1转染前后无显著差异。结论抑制ESRP1表达或RNA识别位点突变可以引起肿瘤中癌基因转录本剪接异常。Objective To investigate the regulatory mechanism of myeloid cell leukemia-1 （MCL1） mRNA splicing mediated by epithelial splicing regulatory protein 1 （ESRP1） in glioma U251 cell lines. Methods Data mining was performed in glioma patient database to find out the key sites from MCL1 sequencing results. Exogenous ESRP1 was over-expressed in U251 cells via pcDNA-ESRP1 vector. The levels of different MCLt isoforms were detected by reverse transcription PCR and Western blotting. Results The mRNA and protein level of endogenous ESRPt was lower in U251 cells than in normal gliocytes. The level of MCL1 isoform 1 was higher, while isoform 2 and 3 were lower in U2.51 cells than in normal gliocytes. Furthermore, MCL1 isoform 1 was reduced and isoform 3 was up-regulated in U251 cells after ESRP1 over-expression. Isoform 2 was unchanged obviously. Finally, MCL1 with the deletion of 801G and 802A sites could not be correctly spliced by ESRPt and no significant difference was seen in the expressions of isoform I and 3 in mutant MCL1. Conclusion Repression of ESRP1 expression or mutation in its RNA recognized sites in tumor results in the abnormality of transcript splicing of oncogenes.

关 键 词：上皮剪接调控蛋白1(ESRP1) 人髓细胞白血病基因1(MCL1) 胶质瘤 U251细胞 剪接异构体 

分 类 号：Q344.14[生物学—遗传学] R392-33[医药卫生—免疫学]