剪接因子SRSF7新mRNA剪接异构体在顺铂处理细胞中的变化  

New m RNA Splicing Isoforms of SRSF7 and Their Change in Cells Treated with Cisplatin

作　　者：马春霞[1] 陈香玲 李明阳[1] 杨渊[1] 刘淑媛 史荔[1] 俞建昆[1] 

机构地区：[1]中国医学科学院医学生物学研究所,云南昆明650118

出　　处：《贵州医科大学学报》2018年第2期154-159,共6页Journal of Guizhou Medical University

基　　金：中国医学科学院医学与健康科技创新工程重大协同创新项目(2016-12M-2-001);国家自然科学基金项目(31570918)

摘　　要：目的:探讨不同梯度浓度顺铂(CDDP)处理不同肺癌细胞后新发现的剪接因子SRSF7 m RNA异构体在同浓度CDDP处理后不同的细胞中的水平变化。方法:采用梯度浓度的CDDP处理肺癌细胞A549(8、18、24、32、40、48及56μmol/L)和H1299(12、24、36、48、60、72及84μmol/L),处理24 h后提取总RNA,进行RT-PCR检测SRSF7 m RNA剪接异构体,并选取NCBI中未确定的异构体进行TA克隆后测序并分析鉴定异构体类型;用56μmol/L的CDDP处理人胚肾细胞293FT,宫颈癌细胞Ca Ski、Si Ha、C33A细胞24 h,提取总RNA后验证新发现的SRSF7 m RNA剪切异构体是否存在这些细胞中。结果:发现3种新的SRSF7 m RNA剪接异构体(分别标记为New1、New2及New3),在肺癌细胞A549、H1299,人胚肾细胞293FT,宫颈癌细胞Ca Ski、Si Ha、C33A细胞等多种肿瘤细胞中均存在,根据NCBI的预测New1,New2,New3异构体均为非编码蛋白的m RNA异构体,随着处理CDDP浓度的增加,其比例明显上升;而编码蛋白的m RNA异构体a和(或)b所占比例明显下降;同时SRSF7的总m RNA水平逐渐降低。结论:新发现的3种非编码的m RNA异构体的变化揭示细胞可能通过SRSF7自身的m RNA选择性剪接来应对DNA损伤。Objective： To explore the changes of mRNA isoforms by gradient （lose of cisplatin and analyze the reasons in A549 and H1299. Methods： Total RNA of the cells were abstracted 2411 after the treatment by cisplatin so as to detect mRNA isofomls of SRSF7 gene by RT-PCR and analyze lhe ratio of the newly discovered mRNA isoforms. The undetermined isomers of NCBI were analyzed. Re- suits： Three new mRNA splicing isoforms （Newl , New2, New3） of SRSF7 were found, which were ubiquitous in tumor cells and were both non-coding mRNA isoforms. With the increase of the concen- tration of eisplatin, their proportion significantly increased, but the ratio of mRNA isoforms that could encode protein was significantly declined. Simuhaneously, the total mRNA level decreased. Conclu- sion： The newly discovered changes in three non-coding mRNA isoforms （Newl , New2, New3 ） reveals that the cells may respond to DNA damage by alternative splicing of SRSFT＇s own mRNA.

关 键 词：顺铂 富含精氨酸丝氨酸的剪接因子7 选择性剪接 异构体 细胞凋亡 

分 类 号：R73-36[医药卫生—肿瘤]