贵州医科大学学报

2018, v.43;No.209(02) 154-159

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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

马春霞;陈香玲;李明阳;杨渊;刘淑媛;史荔;俞建昆;
MA Chunxia;CHEN Xiangling;LI Mingyang;YANG Yuan;LIU Shuyuan;SHI Li;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences;

摘要(Abstract):

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 m RNA isoforms by gradient dose of cisplatin and analyze the reasons in A549 and H1299.Methods:Total RNA of the cells were abstracted 24 h after the treatment by cisplatin so as to detect m RNA isoforms of SRSF7 gene by RT-PCR and analyze the ratio of the newly discovered m RNA isoforms.The undetermined isomers of NCBI were analyzed.Results:Three new m RNA splicing isoforms(New1,New2,New3) of SRSF7 were found,which were ubiquitous in tumor cells and were both non-coding m RNA isoforms.With the increase of the concentration of cisplatin,their proportion significantly increased,but the ratio of m RNA isoforms that could encode protein was significantly declined.Simultaneously,the total m RNA level decreased.Conclusion:The newly discovered changes in three non-coding m RNA isoforms(New1,New2,New3) reveals that the cells may respond to DNA damage by alternative splicing of SRSF7's own m RNA.

关键词(KeyWords): 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
cisplatin;serine/arginine-rich splicing factor 7;selective splicing;isomers;apoptosis

Abstract:

Keywords:

基金项目(Foundation): 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)

作者(Author): 马春霞;陈香玲;李明阳;杨渊;刘淑媛;史荔;俞建昆;
MA Chunxia;CHEN Xiangling;LI Mingyang;YANG Yuan;LIU Shuyuan;SHI Li;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences;

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DOI: 10.19367/j.cnki.1000-2707.2018.02.005

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