人肾癌细胞系ACHN中肿瘤干细胞样细胞的分离及鉴定  

Isolation and characterization of cancer stem-like cells from ACHN renal cancer cells

作　　者：王鹏[1] 王毅东[1] 赵文兵[1] 岳亮[1] 

机构地区：[1]山西中医学院中西医结合医院泌尿外科,太原030013

出　　处：《中国医师杂志》2017年第3期376-379,384,共5页Journal of Chinese Physician

基　　金：山西省自然科学基金项目（2013011046-7）

摘　　要：目的由ACHN人肾癌细胞系中分离肿瘤干细胞样细胞，并鉴定其干细胞相关特性，为进一步研究应用奠定基础。方法采用无血清培养法富集ACHN细胞球，应用MTT法绘制其生长曲线，利用流式细胞仪观察细胞周期，分析细胞表面分子标志物CD133、CD44的表达，利用RT-PCR法分析Oct3/4、Bmi、Nanog、β-catenin干性相关基因的表达，并观察其裸鼠体内成瘤能力。结果ACHN细胞经无血清培养可形成悬浮细胞球，其与ACHN细胞相比，具有更强的体外增殖能力，高表达细胞表面分子标志物CD44，并高表达Oct3/4、Bmi、Nanog、β-catenin等干性相关基因，而且裸鼠体内成瘤能力更强。结论肾细胞癌中存在肿瘤干细胞样细胞，通过无血清培养法可以有效、简便地富集ACHN人肾癌细胞系肿瘤干细胞样细胞，为以肾癌干细胞为靶点的靶向治疗研究奠定了一定基础。Objective To isolate and identify stem-like cells （CSCs） from human ACHN renal cell carcinoma cell line. Methods ACHN cancer stem cells CSCs were enriched by serum-free culture condition. Methyl thiazolyl tetrazolium （MTT） method was performed to draw the growth curve. Flow cytometry was performed to evaluate the expressions of two CSCs markers, CD133 and CD44. Real-time polymerase chain reaction （RT-PCR） was performed to evaluate the expression of stem-related genes （Oct3/4, Bmi, Nanog, and β-catenin）. The tumorigenicity in vivo was also examined. Results ACHN CSCs were formed by sphere-forming culturing. The data demonstrated that there were stronger capacities of proliferation in ACHN tumor spheres than that of ACHN cells. And the expressions of CD133 and CD44 were also significantly enhanced in tumor spheres. The data showed that four genes （ Oct3/4, Bmi, Nanog, and β-catenin） were upregulated in sphere-forming cells. Sphere-forming cells had a higher tumorigenic potential in vivo than incubated cells. Conclusions Human ACHN renal cell carcinoma cell line sphere-forming cells can be enriched by serum-free culture condition and exhibits several characteristic cancer stem cells properties.

关 键 词：肾肿瘤 肿瘤干细胞 

分 类 号：R737.11[医药卫生—肿瘤]