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目的探讨胃癌细胞系中是否存在SP细胞及SP细胞的生物学特征。方法采用荧光活化的细胞分选技术(FACS)从胃癌MKN-45细胞株获得SP细胞,进而分析这些SP细胞有无肿瘤干细胞特性;采用实时定量的RT-PCR检测5个干性相关基因OCT-4,SOX-2,NANOG,CD44和ABCG-2的mRNA在SP和MP细胞中的表达水平,并进行统计学差异分析。结果发现二者之间具有显著差异。进一步使用蛋白印迹法(Western blot)定性分析发现:5个干性相关基因在SP和MP细胞中的蛋白表达水平亦有差异;体内实验结果显示,当SP细胞和MP细胞注射非糖尿病联合重度免疫缺陷(NOD/SCID)的小鼠,SP细胞较MP细胞具有更强的成瘤性。结论综合分析本实验相关结果,可以推测SP细胞具有肿瘤干细胞特性,由MKN-45分选获得的SP细胞是具有干细胞特性的胃癌干样细胞。SP细胞较主群细胞(MP)具有较高的克隆形成率。
Objective To investigate the biological characteristics of SP cells and SP cells in gastric cancer cell lines. Methods SP cells were obtained from gastric cancer cell line MKN-45 by fluorescence activated cell sorting (FACS), and then analyzed for the presence or absence of cancer stem cells in these SP cells. Real-time quantitative RT-PCR was used to detect the expression of 5 stem-related genes OCT- 4, SOX-2, NANOG, CD44 and ABCG-2 mRNA in SP and MP cells, and statistical analysis. The results showed a significant difference between the two. Further qualitative analysis using Western blot showed that there were also differences in the protein expression levels of the five dry related genes in SP and MP cells. In vivo experiments showed that when SP cells and MP cells were injected with non-diabetic severe immunosuppression Deficient (NOD / SCID) mice, SP cells have more tumorigenicity than MP cells. Conclusion According to the results of this experiment, we can speculate that SP cells have tumor stem cell characteristics. The SP cells obtained from MKN-45 cells are stem cell-like stem-like cells. The SP cells have a higher clonogenic rate than the main group cells (MP).