作　　者： (马春霞）; (杨梦莉）; (李明阳）; (杨渊）; (陈香岭）; (俞建昆）;

机构地区： 中国医学科学院＆北京协和医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》 2017年第8期879-884,892,共7页

摘　　要： 目的:观察不同浓度顺铂处理后肺癌细胞中剪接因子(SRSF1)mRNA异构体的变化,并寻找其中发挥主要抗凋亡作用的异构体。方法:使用不同浓度的顺铂处理肺癌细胞系A549和H1299后提取总RNA,设计SRSF1基因不同的引物(F1∶R1、F1∶F2及F2∶R1)进行RT-PCR以检测其mRNA剪接异构体的变化。结果:在A549和H1299细胞中检测到了5种异构体(Isoform-a、Isoform-b、Isoform-d、Isoform-e、Isoform-f),且主要表现为随着顺铂浓度的升高,非编码蛋白的mRNA异构体的比例明显上升,主要是Isoform-d的上升;而编码蛋白的mRNA异构体所占比例明显下降,主要是Isoform-a下降;同时SRSF1的总mRNA水平也是逐渐降低的。结论:在顺铂处理下,肺癌A549和H1299细胞中的SRSF1的转录水平被下调,其mRNA选择性剪接向下调编码蛋白的mRNA异构体的方向转变,造成编码蛋白的mRNA异构体降低的主要原因为Isoform-a,该异构体可能是SRSF1基因发挥抗细胞凋亡作用的主要异构体。 Objective： To detect the changes of splicing factor SRSF1 mRNA isoforms in lung cancer cells treated with cisplatin and search for isoforms that play a major role in anti apoptotic activity.Methods： After using cisplatin of different concentrations treating lung cancer cell lines A549 and H1299,the total RNA was extracted and the different primers（ F1 ∶ R1、F1 ∶ F2 及 F2 ∶ R1） of SRSF1 gene were designed to conduct RT-PCR and test the change of mRNA splicing variants. Results： Under cisplatin treatment,5 mRNA isoforms of SRSF1 in cell A549 and H1299 were detected,and with the increase of cisplatin concentration,mRNA isomers of non encoding proteins increased,especially Isoform-d,the proportion of mRNA encoding protein isomer significantly decreased,especially Isoforma. Meanwhile the level of total mRNA in SRSF1 also decreased gradually. Conclusion： Under the treatment of cisplatin,transcriptional level of SRSF1 in lung cancer A549 and H1299 cells was reduced,and mRNA alternative splicing changed from isoform encoding protein to non-coding one.

关 键 词： 顺铂 剪接因子 剪接 选择性剪接 异构体 细胞凋亡