鼻咽癌干细胞增殖特征及细胞周期相关蛋白表达水平研究  

Proliferation characteristics and cell cycle-related protein expression in nasopharyngeal carcinoma stem cells

作　　者：何兰 范婧莹 史红健 周芳亮 罗晶婧 何迎春 蒋益兰 廖端芳 HE Lan;FAN Jingying;SHI Hongjian;ZHOU Fangliang;LUOJingJing;HE Yingchun;JIANG Yilan;LIAO Duanfang(Graduate School,Hunan University of Chinese Medicine,Changsha,Hunan,41020&China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Traditional Chinese Medicine,Changsha,Hunan,410208,China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Traditional Chinese Medicine and Visual Function Protection Engineering and Technological Research Center,Changsha,Hunan,410208,China)

机构地区：[1]湖南中医药大学研究生院,湖南长沙410208 [2]中医药防治眼耳鼻咽喉疾病湖南省重点实验室,湖南长沙41020 [3]湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心,湖南长沙410208

出　　处：《中国耳鼻咽喉头颈外科》2019年第10期526-530,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：国家自然科学基金面上项目(81573721、81874408);湖南省自然科学基金项目(2019JJ40216、2017JJ3246);湖南省教育厅科研基金项目(16C1212)

摘　　要：目的研究鼻咽癌干细胞的增殖特征并初步探讨其分子机制。方法采用无血清悬浮培养法从鼻咽癌细胞CNE2和5-8F获得鼻咽癌干细胞CNE2-SC、5-8F-SC,分别采用细胞存活率分析计数器、细胞平板克隆形成试验观察鼻咽癌干细胞的增殖特性和克隆形成能力,Western blot法检测细胞周期相关蛋白表达水平。结果鼻咽癌干细胞比其来源的鼻咽癌细胞增殖速度显著降低,差异有统计学意义(P<0.05)。鼻咽癌干细胞克隆形成率显著高于其来源鼻咽癌细胞(P<0.01),但肿瘤干细胞克隆显著小于其来源细胞。与普通鼻咽癌细胞比较,鼻咽癌干细胞的细胞周期蛋白CylinD1、CylinD3及细胞周期调控蛋白CDK2、CDK4、CDK6的表达水平显著下调(P<0.01)。结论鼻咽癌干细胞比其来源的鼻咽癌细胞生长增殖缓慢,可能与其细胞周期蛋白和周期蛋白依赖激酶表达水平下调有关。OBJECTIVE To study the proliferation characteristics of nasopharyngeal carcinoma stem cells and to explore their molecular mechanisms. METHODS Nasopharyngeal carcinoma stem cells CNE2-SC and 5-8 F-SC were obtained by serum-free suspension culture with CNE2 and 5-8 F cells. And on the basis of this, cell counting instrument and colony formation assay were used to check the proliferation of stem cells, and Western blot test were used to test protein expression levels. RESULTS The growth rate of nasopharyngeal carcinoma stem cells was significantly lower than that of which the nasopharyngeal carcinoma stem cells grown from, and the difference was statistically significant(P<0.05). After 14 days of implantation, colony forming efficiency(CFE) of nasopharyngeal carcinoma stem cells was higher than that of nasopharyngeal carcinoma(P<0.01). Compared with nasopharyngeal carcinoma cells, the expression levels of CylinD1, CylinD3, CDK2, CDK4 and CDK6 in nasopharyngeal carcinoma stem cells were obviously downregulated(P<0.01). CONCLUSION The nasopharyngeal carcinoma stem cells grow at a relatively slow rate compared to that of nasopharyngeal carcinoma cells which the nasopharyngeal carcinoma stem cells grown from, that may be related to low expression levels of cyclin proteins and cell cycle related proteins.

关 键 词：细胞增殖 干细胞 细胞周期蛋白类 周期蛋白依赖激酶 克隆形成力 

分 类 号：R73[医药卫生—肿瘤]