常 诚沈丽萍张学清李 琳李孝鑫王治东周平坤

1（南华大学公共卫生学院 衡阳 421000）

2（军事医学科学院放射与辐射医学研究所 北京 100850）



辐射诱导分子lnc-RI稳定敲低细胞系建立及其功能

常 诚1,2沈丽萍2张学清2李 琳2李孝鑫2王治东2周平坤1,2

1（南华大学公共卫生学院 衡阳 421000）

2（军事医学科学院放射与辐射医学研究所 北京 100850）

摘要制备lnc-RI干涉表达的慢病毒颗粒，建立lnc-RI稳定下调的HeLa细胞系，探索lnc-RI对细胞辐射敏感性的作用。首先将lnc-RI特异性干涉序列插入干涉表达载体pGLV2中，并制备慢病毒颗粒；然后，将慢病毒颗粒感染HeLa细胞，建立lnc-RI稳定下调的细胞系，并通过RT-PCR检测不同干涉序列对lnc-RI表达的干涉效果；最后，通过平板克隆形成实验分析lnc-RI表达下调对HeLa细胞放射敏感性的影响。结果显示，成功制备了携带lnc-RI干涉序列的慢病毒颗粒，并成功感染HeLa细胞，通过RT-PCR实验鉴定获得两株lnc-RI稳定敲低细胞系。克隆形成实验发现，与对照相比，lnc-RI敲低的HeLa细胞系的放射敏感性增加。因此，初步证实lnc-RI表达下调增加HeLa细胞放射敏感性。

关键词长链非编码RNA，lnc-RI，慢病毒，RNA干涉，电离辐射

wangzhidong1977@126.com

Supported by the National Natural Science Foundation of China (No.81573083 and 81402631)

First author: CHANG Cheng, female, was born in June 1987 and graduated from North China University of Science and Technology in 2012. Now she is a master candidate of University of South China, majoring in public health and preventive medicine and focusing on health toxicology. E-mail: 1109308002@qq.com

professor, E-mail: wangzhidong1977@126.com

Received 17 September 2015; accepted 20 October 2015

Establishment of stable knock-down cell lines and function study of lnc-RI, a radiation-induced gene

CHANG Cheng1,2SHEN Liping2ZHANG Xueqing2LI Lin2LI Xiaoxin2WANG Zhidong2ZHOU Pingkun1,2

1(College of Public Health, University of South China, Hengyang 421000, China)
2(Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China)

ABSTRACT To prepare the lentivirus granules interfering lnc-RI and establish HeLa cell lines consistently down-regulating lnc-RI, as well as discover its effects on radiation damage, the lentivirus granules bearing various interfering sequences were prepared by lnc-RI carrying specific interfering sequences in to interfering plasmid pGLV2. The estable cell lines were established after infecting HeLa cells with lentivirus granules. RT-PCR was employed to determine the inhibitory effects of various interfering sequences on lnc-RI expression. DNA damage of the established estable cell lines were induced by ionizing radiation sensitivity of the established cell lines which weredetermined by colony formation assay. The results showed that the lentivirus granules bearing various interfering sequences were successfully prepared. The estable HeLa cell lines infected by lentivirus were obtained and lnc-RI expression levels was down-regulated. The results of colony formation assay showed that radiation sensitivity of the estable cell lines increased when treated with each dose gamma ray irradiation.

KEYWORDS Long non-coding RNA, lnc-RI, Lentivirus, RNA interference, Ionizing radiation

CLC TL71

长链非编码RNA (Large non-coding RNA, lncRNA)是一类转录本长度大于200个核苷酸、绝大部分没有长阅读框架、没有编码蛋白质的功能、但像mRNA一样具有帽式结构和poly A尾巴的基因转录产物[1]。随着基因组技术的发展，发现在人类基因组中大部分lncRNA存在转录现象，但与编码蛋白质的基因序列相比，其表达水平相对较弱[2]。lncRNA自身的表达水平不仅受到转录及转录后调控机制的严密调节[3]，它还……