贵州医科大学学报

2016, v.41;No.194(11) 1279-1284

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顺铂处理后几种hTERT mRNA剪接异构体的变化
Several Splice Isoforms of hTERT mRNA and Its Change in Cells after Treatment with Cisplatin

杨梦莉;陈香岭;隋旭;廉亚茹;马春霞;姚宇峰;史荔;马千里;俞建昆;
YANG Mengli;CHEN Xiangling;SUI Xu;LIAN Yaru;MA Chunxia;YAO Yufeng;SHI Li;MA Qianli;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;Kunming University of Science and Technology;Department of Thoracic Surgery,the Third Affiliated Hospital of Kunming Medical University;

摘要(Abstract):

目的:利用顺铂处理肺癌、宫颈癌细胞和人胚肾细胞,检测人类端粒酶逆转录酶催化亚单位(hTERT)mRNA的选择性剪接是否发生变化。方法:将培养的肺癌A549和H1299细胞,宫颈癌C33A、SiHa和Ca Ski细胞及人胚肾293FT细胞分为实验组和对照组,用顺铂处理实验组细胞,DMSO处理对照组细胞;设计hTERTα+β+、α+β-、α-β+、INS3及DEL[e2]引物,提取两组细胞的总RNA,利用RT-PCR检测hTERT mRNA剪接异构体表达。结果:与对照组细胞相比,随着顺铂浓度的增加,肺癌A549和H1299细胞,宫颈癌C33A、SiHa和Ca Ski细胞以及人胚肾293FT细胞中hTERTα+β-和INS3异构体比例升高,而DEL[e2]异构体变化不明显。结论:在顺铂引起的DNA损伤情况下,细胞可利用hTERT mRNA选择性剪接调控机制进行调节,通过改变不同异构体的量和所占比例,从而调节端粒酶功能活性。
Objective: To determine whether there is any change of alternative splicing of h TERT mRNA in lung cancer and cervical cancer cells and human embryonic kidney cells after cisplatin treatment,different dose of cisplatin were used to treat these two types of cell line. Methods: Cultivated lung cancer A549 and H1299 cells,cervical cancer C33 A,Si H aand Ca Ski cells and HEK 293 FT cells were divided into experiment group and control group; using cisplatin to treat experiment group cells and DMSO to treat control group cells; h TERT α + β +,α + β-,α- β +,INS3,DEL[e2]alternative splicing sites were selected to design primers. Total RNA of treated cells and control cells after treatment with cisplatin were extracted and RT-PCR was used to test the spliced transcripts,and their ratios of h TERT mRNA in cancer cells. Results: The results indicated that the ratio of the h TERT α +β- and INS3 isoform increased with the increase of the concentration of cisplatin; but the change of DEL [e2] was not obvious. Conclusion: The activity of telomerase may be regulated through the expression and ratio of different alternative splicing isoforms of h TERT when DNA impairment.

关键词(KeyWords): 肺肿瘤;宫颈肿瘤;人胚肾细胞;细胞,培养的;人类端粒酶逆转录酶催化亚单位;顺铂
lung neoplasm;cervical neoplasm;human embryonic kidney cells;cell,cultured;human telomerase reverse transcriptase;cisplatin

Abstract:

Keywords:

基金项目(Foundation): 中国医学科学院病原生物学研究所基本科研业务费项目(2012IPB107);; 高等学校博士学科点专项科研基金(新教师类)(20111106120056)

作者(Author): 杨梦莉;陈香岭;隋旭;廉亚茹;马春霞;姚宇峰;史荔;马千里;俞建昆;
YANG Mengli;CHEN Xiangling;SUI Xu;LIAN Yaru;MA Chunxia;YAO Yufeng;SHI Li;MA Qianli;YU Jiankun;Institute of Medical Biology,Chinese Academy of Medical Sciences & Peking Union Medical College;Kunming University of Science and Technology;Department of Thoracic Surgery,the Third Affiliated Hospital of Kunming Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2016.11.009

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