| 吉 晨,吴玉梅,李秀楠.宫颈癌细胞辐射敏感性与IER5基因的关系[J].肿瘤学杂志,2017,23(4):305-311. |
| 宫颈癌细胞辐射敏感性与IER5基因的关系 |
| The Relationship Between Radiosensitivity of Cervical Cancer Cells and IER5 Gene |
| 投稿时间：2016-04-25 |
| DOI：10.11735/j.issn.1671-170X.2017.04.B011 |
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| 中文关键词: ER5基因 SiHa细胞 细胞系构建 辐射敏感性 宫颈肿瘤 |
| 英文关键词:IER5 gene SiHa cells cell lines construction radiosensitivity cervical neoplasms |
| 基金项目:国家自然科学基金资助项目（81272888） |
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| 中文摘要: |
| 摘 要：［目的］ 通过RNA干扰技术致基因沉默和基因过表达的方法研究IER5基因与宫颈癌SiHa细胞系辐射敏感性的相关性，探讨IER5基因不同表达的细胞系对辐射敏感性的差异。［方法］ 构建IER5沉默及过表达的真核表达载体，转染细胞并筛选出稳定表达的细胞系，经Western Blot和Real-time PCR鉴定成功。观察IER5基因不同表达的3种细胞系的形态；对细胞系进行4Gy的辐射后使用CCK-8检测细胞生长曲线；对细胞系进行10Gy的辐射后使用流式细胞仪检测细胞凋亡情况；对细胞系进行4Gy的辐射后使用流式细胞仪检测不同时间点细胞周期的情况。［结果］ IER5基因沉默使SiHa细胞略有增大，照射后的细胞增殖速度加快；而其过表达可使SiHa细胞缩小呈圆形，并且细胞增殖速度有所减慢。IER5沉默导致辐射诱导的SiHa细胞凋亡率降低，过表达可促进辐射诱导的SiHa细胞凋亡，其凋亡在照射后48h最为显著。照射后细胞周期的G2-M期阻滞效应随IER5表达含量的降低而升高，IER5基因沉默促进细胞的增殖分裂。［结论］ IER5基因与宫颈癌SiHa细胞的增殖、凋亡和细胞周期的变化及对辐射的反应有明显相关性，IER5基因可能可提高宫颈癌细胞对放射的敏感性。 |
| 英文摘要: |
| Abstract：［Objective］ Using RNA interference and gene overexpression method to make IER5 gene silence and overexpression，in order to research the correlation between IER5 gene with cervical cancer cells SiHa’s radiation sensitivity，to explore the differences of radiotherapy sensitivity in different IER5 gene expression of cell lines.［Methods］ IER5 silence and overexpression eukaryotic expression vectors was established，transfected SiHa cells and select stably expressing cell lines，and verified the cell line by Western Blot and Real-time PCR turns out the cell line to be successfully established. Observing the change in cell morphology of three kinds of cell lines；Using CCK-8 to measure cell growth curve after 4Gy of radiation；Using the flow cytometry to detect cell apoptosis after 10Gy of radiation；Using the flow cytometry to detect cell cycles after 4Gy of radiation in different timing.［Results］ IER5 gene silencing SiHa cells increased in volume slightly，and the speed of cell proliferation after radiation had increased. And IER5 gene overexpression could make SiHa cells shrink assumes as circle，and cell proliferation had slowed down. IER5 gene silencing resulted in the decrease of SiHa cells apoptosis rate induced by radiation，and IER5 gene overexpression could promote the SiHa cells apoptosis rate induced by radiation. The result presented most significant apoptosis rate appears in 48-hour after radiation. The G2-M phase retardation effect in cell cycle after radiation was promoted along with the decreasing of IER5 expression，which indicates IER5 gene silencing can promote cell proliferation. ［Conclusion］IER5 gene has significant correaltion with cervical cancer SiHa cell lines’ proliferation，apoptosis，cell cycles and response to radiation. IER5 gene can improve the sensitivity of cervical cancer cells to radiation. |
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