人肺癌细胞系NCI-H1650中肿瘤干细胞的分离及鉴定  

Isolation and identification of lung cancer stem cells from NCI-H1650 cell line

作　　者：丁曼[1] 张岸梅[1] 朱波[1] 

机构地区：[1]第三军医大学新桥医院肿瘤科/全军肿瘤诊治研究所,重庆400037

出　　处：《重庆医学》2015年第12期1588-1591,共4页Chongqing medicine

基　　金：国家自然科学基金青年科学基金资助项目(81302025)

摘　　要：目的在人肺癌细胞系NCI-H1650中分离出肺癌干细胞及鉴定。方法将NCI-H1650细胞在含人胰岛素、重组人表皮生长因子(EGF)、重组人碱性成纤维细胞生长因子(bFGF)、牛血清清蛋白(BSA)的无血清培养基中连续培养,用紫杉醇刺激诱导干细胞,通过流式细胞术、免疫荧光的方法分析肺癌干细胞标志物CD133和CD326的表达;采用RT-PCR和Western blot的方法检测干细胞相关标记物Oct4、Nanog和Sox-2的表达。结果经无血清连续培养结合紫杉醇诱导的NCI-H1650细胞呈球形悬浮生长,相比于有血清培养的NCI-H1650细胞,其干细胞标志物CD133和CD326高表达,干细胞相关标志物Oct4、Nanog和Sox-2表达水平提高(P<0.05)。结论采用体外无血清连续培养结合紫杉醇刺激诱导NCI-H1650细胞能有效分离出其中干细胞。Objective To isolate and identify the lung cancer stem like cells from the cell line NCI-H1650. Methods The NCI-H1650 cells were continuously cultured in serum-free medium with human insulin, recombinant epidermal growth factor （EGF）, recombinant basic fibroblast growth factor （bFGF） and bovine serum albumin（BSA） ,and paclitaxel was adopted to induce the stem cells. Flow cytometry and immunofluorescent staining were used to analyze the expression levels of lung cancer stem cell marker CD133 and CD326. The expressions of stem cell related marker Oct4, Nanog and Sox-2 were detected by RT-PCR and West- ern blot. Results The NCI-H1650 cells continuously cultured by serum-free medium and induced by combining paclitaxel induction showed the spherical suspension growth. Compared with NCI-H1650 cells cultured by serum,CD133 and CD326 had significantly high expression and the expression levels of Oet4,Nanog and Sox-2 were significantly increased （P〈0.05）. Conclusion Adopting the serum-free culture in vitro combined with paclitaxel for inducing NCI-H1650 cells can effectively isolate the stem cells among them.

关 键 词：肺肿瘤 肿瘤干细胞 培养基 无血清 

分 类 号：R734.2[医药卫生—肿瘤]