辐射对小鼠皮肤成纤维细胞株L929增殖及CCN第三军医大学学报.doc

辐射对L929细胞生长及CCN1基因表达的影响 万莹华1，司维柯1，杜业军1，李招权1，潘静1，赵宸1，李军1，粟永萍2 （基金项目：全军十一五国际合作项目基金资助（06H034），创伤、烧伤、复合伤国家重点实验室开放课题作者简介：万莹华，女，1983-4，江西南昌，硕士研究生，助教，Tel:(023E-mail： xiaokeailingxi@.通信 基金项目：全军十一五国际合作项目基金资助（06H034），创伤、烧伤、复合伤国家重点实验室开放课题 作者简介：万莹华，女，1983-4，江西南昌，硕士研究生，助教，Tel:(023E-mail： xiaokeailingxi@. 通信作者：司维柯，Tel：023Email： weikesi2004@；粟永萍，Tel：023Email： suyping@. 提 要：目的：通过观察辐射对小鼠皮肤成纤维细胞系L929生长及CCN1（能否首先说明何为CCN1基因？）表达的影响，探讨CCN1与辐射损伤的关系。方法：体外培养L929细胞系，4Gy60Coγ射线照射为辐射组，并设对照组，MTT法测细胞增殖变化，平板克隆形成试验测克隆形成能力，流式细胞术测细胞周期，RT-PCR法及免疫细胞化学法检测L929细胞中CCN1的表达情况。结果：辐射组细胞2天增殖抑制明显（P＜0.01），克隆形成也明显受到抑制（P＜0.05）。辐射后24小时，细胞出现G2期停滞。辐射后6小时L929 CCN1的mRNA水平表达明显降低，至24小时开始回调，辐照后12-48小时，细胞CCN1蛋白水平的表达明显下降（P＜0.01）。结论：因辐照生长受到抑制的小鼠成纤维细胞L929，其CCN1基因的表达水平下调，提示CCN1基因的表达水平低下是辐射导致细胞生长抑制的因素之一。 关键词：辐射，电离；成纤维细胞系，L929；CCN1/Cyr61 中图分类号： The effects of radiation on growth and CCN1 expression of L929 cells WAN Ying-hua1, SI Wei-ke1,DU Ye-jun1, LI Zhao-quan1, PAN Jing1, ZHAO Chen1, LI Jun1, SU Yong-ping2（Department of Clinical Hematology, Third Military Medical University, Chongqing 400038；China;Institute of Combined Injury, College of Preventive Medicine, State Key Laboratory of Trauma, Burns and Combined Injury, Third Military Medical University, Chongqing 400038, China）（单位标注不规范，请参照其他相关文章） Abstract：Objective：To observe the effects of radiation on growth and CCN1 expression of L929 cells and investigate the relationship between CCN1 expression and radiation injury. Method：L929 cells were cultured in vitro，which were divided into two groups：irradiated cells with 4 Gy γ-irradiation as radio-group, untreated cells as control group. The proliferation of cells was test by MTT assay and plate colony formation experiment. Flow cytometry was utilized to quantify the cell cycle distribution. CCN1 protein and mRNA were determined by immunocytochemistry (ICC) and RT-PCR respectively. Result：Significant inhibition of proliferation (P＜0.01＝ and colony formation (P＜0.05＝ was found in cells