人胃癌细胞株SGC-7901的克隆形态观察及含肿瘤干细胞克隆的鉴定  

Morphological observation of human gastric cancer cell SGC-7901 clones and identification of gastric cancer stem cells

作　　者：杨宏琼[1,2] 周志华[3] 张尤历[1] 徐岷[1] 徐萍[1] 吴莺[1] 王银环[1] 

机构地区：[1]江苏大学附属医院消化内科,镇江212001 [2]常州市第一人民医院干部病房,213003 [3]解放军第一0一医院病理科

出　　处：《中华肿瘤杂志》2013年第3期164-169,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金（81000964）

摘　　要：目的动态观察人胃癌SGC-7901细胞的克隆在形态上的差异，比较不同克隆细胞的裸鼠体内成瘤能力，并鉴定含胃癌干细胞的克隆。方法采用克隆形成实验观察胃癌SGC-7901细胞克隆的形态并分类，计算克隆形成率和每种克隆的比例。采用免疫荧光技术和Westernblot法检测不同克隆细胞中CIM4和CDX2的表达。分离不同形态的克隆，检测其自我更新能力。将不同形态的克隆细胞分别接种于裸鼠皮下，观察各组克隆细胞的体内成瘤能力。结果SGC-7901细胞可形成3种克隆形态，即紧密型克隆、中间型克隆和松散型克隆。SGC-7901细胞总的克隆形成率为（9．80±1．07）％，其中紧密型、中间型和松散型克隆所占的比例分别为10．2％、56．0％和33．8％。免疫荧光法检测的结果显示，CIM4在紧密型克隆细胞中的表达明显强于中间型和松散型克隆，而CDX2在紧密型克隆细胞中的表达最弱。Western blot法检测结果与免疫荧光法检测的结果一致。紧密型克隆细胞具有很强的自我更新能力，在裸鼠体内的成瘤能力也较强，接种10^3和10^4个细胞的成瘤率分别为75．0％和100％。结论SGC-7901细胞的克隆在形态和分化程度上存在差异，其中紧密型克隆可能富含胃癌干细胞。Objective To dynamically investigate the morphology of human gastric cancer SGC- 7901 cell clones, and then compare the tumorigenic ability of different clones in order to identify the tumor stem cell clones. Methods Clones derived from gastric cancer SGC-7901 cells were assessed by morphological observation, and the clone formation rate and proportion of each clone were calculated. The expression of CD44 and CDX2 in different clones was detected by immunofluorescence microscopy and Western blot. Furthermore, different clones were isolated and cultured, and their self-renewal property was assayed. Cells of different clones were subcutaneously inoculated into nude mice and the tumorigenic ability of each group was determined. Results Clones derived from gastric cancer SGC-7901 cells had three types, i.e. clones of tight, transitional and loose types. The total clone formation rate was （9.80 ± 1.07） %, and the proportion of tight,transitional and loose type clones was 10.2% ,56.0% and 33.8%, respectively. The results of immunofluorescence microscopic examination showed that the signal of CD44 was significantly stronger in the tight clones than in the transitional and loose clones, however, the signal of CDX2 was weakest in the tight colonies. The results of Western blot were consistent with that of immunofluorescence microscopic observation. SGC-7901 cells of tight clones possessed strong ability of self-renewal and in vivo tumorigenicity in the nude mice. Conclusion SGC-7901 cell clones vary in morphology and differentiation, and the tight type clones may include rich gastric cancer stem cells.

关 键 词：胃肿瘤 肿瘤干细胞 克隆 CD44 CDX2 裸鼠 

分 类 号：R735[医药卫生—肿瘤]