X射线修复交叉互补组基因1沉默后人支气管上皮细胞生物学性状研究  

Study on biological characteristics of human bronchial epithelial cells with silencing of XRCC1 gene

作　　者：方道奎[1] 刘移民[2] 胡大林[3] 袁建辉[1] 庄志雄[1] 

机构地区：[1]深圳市疾病预防控制中心,广东深圳518055 [2]广州市职业病防治院,510420 [3]广东医学院,523770

出　　处：《中国职业医学》2012年第1期5-7,12,共4页China Occupational Medicine

基　　金：国家自然科学基金资助项目(81060241);广东省科技厅社会发展与基础研究立项项目(2010)

摘　　要：目的探讨RNA干扰(RNAi)技术使人X射线修复交叉互补组基因1(XRCC1)沉默后人支气管上皮细胞(HBE)的生物学性状变化。方法研究设实验组(转染重组质粒的HBE)、空载体组(转染空载体的HBE)、正常对照组(正常HBE)。XRCC1干扰后的HBE的生物学性状分析运用载体介导的RNAi技术抑制XRCC1在HBE中的表达;在光学显微镜下观察实验组和正常对照组细胞的形态学特征;对实验组、空载体组及正常对照组绘制细胞生长曲线,分析细胞倍增时间,运用流式细胞技术分析细胞周期;以汉丽埃塔(HeLa)细胞作阳性对照,采用软琼脂糖恶性程度鉴定实验分析实验组细胞的恶性程度。结果实验组细胞体积稍有增大,细胞内颗粒物明显增多;实验组、空载体组及正常对照组细胞的生长曲线均呈现相似的"S"型;实验组、空载体组及正常对照组细胞的倍增时间相近,分别为0.70、0.83、0.74 d;3组细胞的G1期、G2期和S期分布相近;实验组细胞未见在双层软琼脂糖中生长出细胞集落,而阳性对照细胞集落明显。结论 XRCC1基因缺陷对HBE生物学性状的短期影响不明显。Objective To explore the biological characteristics of human bronchial epithelial cells（HBE） with RNA interference（RNAi） for X-ray repair cross complementing group 1（XRCC1）gene. Methods Experimental group（HBE of transfection restructuring plasmid）,empty vector group（HBE of transfection empty）,and normal control group（normal HBE） were designed.Vector-mediated RNAi was used to inhibit the expression of XRCC1 in HBE.For experimental group and normal control group,morphological characteristics were analyzed under light microscope;for experimental group,empty vector group and normal control group,growth curves were drawn,doubling generation time was analyzed and cell cycle was studied with technology of flow cytomertry（FCM）.HeLa cell was chosen as positive control group and malignancy degree for experimental group was analyzed with soft agar assay. Results In the experimental group,apparently increased cell granulations and slightly increased cell volume were found.Each group presented similar S-shaped growth curve.The doubling generation time of the experimental group,empty vector group,normal control group were 0.70,0.83 and 0.74 days respectively,while the cell cycle distribution of G1,G2 and S in 3 groups appeared similar.In soft agar assay,cell colony was not found in the experimental group,but found in the positive control group. Conclusion There is no apparent short-term influence on biological characteristics of human bronchial epithelial cells with silencing of XRCC1 gene.

关 键 词：人X射线修复交叉互补组基因1 RNA干扰 人支气管上皮细胞 生物学性状 

分 类 号：Q555[生物学—生物化学] R994[医药卫生—毒理学]