亚砷酸钠诱导HaCaT细胞恶性转化实验  

Malignant Transformation of HaCaT Cells Induced by Sodium Arsenite

作　　者：杨桥樱[1] 黄世文[1] 吴元桢[1] 肖雪[1] 郭菲烨 郭松超[1] 

机构地区：[1]广西医科大学公共卫生学院营养与食品卫生学教研室,南宁市530021 [2]日本三重大学医学研究生院环境和分子医学部

出　　处：《广西医学》2013年第5期530-533,544,共5页Guangxi Medical Journal

基　　金：广西科学研究与技术开发计划项目(0816004-1)

摘　　要：目的探讨低浓度亚砷酸钠(NaAsO2)对人永生化但无致癌性的皮肤角质细胞株(HaCaT)的影响。方法 (1)细胞培养:HaCaT细胞株常规培养,隔日换液,4 d传代;(2)细胞毒性实验:MTT法检测细胞毒性,确定最佳染毒剂量;(3)细胞转化实验:HaCaT细胞在含0.05 ppm NaAsO2培养液中连续传代培养40代(约160 d),细胞培养期间,倒置显微镜下观察每代细胞形态的变化并采集图片,MTT法检测细胞增殖能力,半固体琼脂实验检测细胞克隆形成率。结果 NaAsO2处理细胞形态学、增殖能力及非停泊依赖性能力均发生变化,主要表现为多核巨细胞,倍增时间缩短,在半固体琼脂培养基上形成集落及克隆形成率明显增加。结论 NaAsO2处理的细胞发生恶性转化,砷诱导体外细胞恶性转化模型建立成功。Objective To investigate the effect of low-dose NaAsO2 on human immortalized non-carcinogenic HaCaT. Methods （1） Cell culture：HaCaT routine culture medium was changed every other day, and passaged on the forth day. （2） Cytotoxicity test： Methyl thiazolyl tetrazolium（MTT） assay was used to detect the cellular toxicity ,and the best exposure dose was determined. （3） Cell transformation assay ： HaCaT cells successively subcultured 40 passages in the culture medium of 0.05 ppm NaAs02. During cell culture, we observed each passage＇s morphological changes and collected pictures with an inverted microscope;MTl＇ assay was used to detect cell proliferation,and semi-solid agar assay was adopted to detect colony formation. Results The cells treated with NaAsO2 were changed in morphology,proliferation and non-parked dependent ability,which mainly presented as multinucleated giant eells, shortened doubling time, significantly increased form colonies in the semi-solid agar medium and colony formation. Conclusion The cells treated with NaAsO2 present a malignant transformation, and the model of arsenic-induced malignant cell transformation in vitro is established.

关 键 词：亚砷酸钠 皮肤角质细胞 恶性转化 体外细胞转化 

分 类 号：R995[医药卫生—毒理学]