辐射诱导分子lnc-RI稳定敲低细胞系建立及其功能
Establishment of stable knock-down celllines and function study of lnc-RI,a radiation-induced gene
- 辐射研究与辐射工艺学报 2016年34卷第1期 页码：10203
- 作者机构：
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
- 作者简介：
常诚，女，1987年6月出生，2012年毕业于华北理工大学，现为南华大学在读研究生，公共卫生与预防医学专业，研究方向卫生毒理学，E-mail: 1109308002@qq.com
王治东，副研究员，博士，E-mail: wangzhidong1977@126.com
周平坤，博士生导师，研究员，E-mail: zhoupk@bmi.ac.cn
- 基金信息：国家自然科学基金面上项目(81573083)和国家自然科学基金青年项目(81402631)资助
DOI：10.11889/j.1000-3436.2016.rrj.34.010203
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常诚, 沈丽萍, 张学清, 等. 辐射诱导分子lnc-RI稳定敲低细胞系建立及其功能[J]. 辐射研究与辐射工艺学报, 2016,34(1):10203.
Cheng CHANG, Liping SHEN, Xueqing ZHANG, et al. Establishment of stable knock-down celllines and function study of lnc-RI,a radiation-induced gene[J]. Journal of Radiation Research and Radiation Processing, 2016,34(1):10203.
常诚, 沈丽萍, 张学清, 等. 辐射诱导分子lnc-RI稳定敲低细胞系建立及其功能[J]. 辐射研究与辐射工艺学报, 2016,34(1):10203. DOI： 10.11889/j.1000-3436.2016.rrj.34.010203.
Cheng CHANG, Liping SHEN, Xueqing ZHANG, et al. Establishment of stable knock-down celllines and function study of lnc-RI,a radiation-induced gene[J]. Journal of Radiation Research and Radiation Processing, 2016,34(1):10203. DOI： 10.11889/j.1000-3436.2016.rrj.34.010203.
摘要
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。
Abstract
ABSTRACT, To prepare the lentivirusgranules interfering lnc-RI and establish HeLa cell lines consistentlydown-regulating lnc-RI, as well as discover its effects on radiation damage, the lentivirusgranules bearing various interfering sequences were prepared by lnc-RI carryingspecific interfering sequences in to interfering plasmid pGLV2. The establecell lines were established after infecting HeLa cells with lentivirusgranules. RT-PCR was employed to determine the inhibitory effects of variousinterfering sequences on lnc-RI expression. DNA damage of the established establecell lines were induced by ionizing radiation sensitivity of the establishedcell lines which weredetermined by colony formation assay. The results showed that the lentivirusgranules bearing various interfering sequences were successfully prepared. Theestable HeLa cell lines infected by lentivirus were obtained and lnc-RI expression levels wasdown-regulated. The resultsof colony formation assay showed that radiation sensitivity of the estable celllines increased when treated with each dose gamma ray irradiation.
关键词
长链非编码RNAlnc-RI慢病毒RNA干涉电离辐射
Keywords
Longnon-coding RNAlnc-RILentivirusRNA interferenceIonizing radiation
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