胃癌细胞系SGC-7901肿瘤干细胞样细胞的分离和鉴定

Identification of Gastric Cancer Stem-like Cells in Cell Line SGC-7901

目的探讨人胃癌细胞系SGC-7901中鉴定胃癌干细胞样细胞。方法采用无血清培养基培养胃癌细胞系SGC-7901,得到悬浮生长状态的细胞球;流式细胞术分别检测贴壁胃癌细胞和悬浮细胞球中Side Population(SP)侧群细胞的含量;NOD/SCID鼠体内异位移植成瘤实验检测成瘤能力;基因芯片技术筛选悬浮细胞球与贴壁细胞差异表达的基因;实时荧光定量PCR验证基因芯片结果。结果 (8.00±0.54)%SGC-7901细胞在无血清培养基中能增殖并形成可连续传代的细胞球;悬浮细胞球中SP细胞百分含量高达(12.34±1.01)%,贴壁细胞情况中SP仅占(0.62±0.05)%;NOD/SCID小鼠成瘤实验显示,悬浮球细胞组致瘤能力高于贴壁细胞组;基因芯片结果显示,肿瘤球高表达与干细胞相关的基因ABCG2、HES1和KRT153。实时荧光定量PCR方法检测结果与基因芯片结果基本一致。结论胃癌细胞系SGC-7901中存在肿瘤干细胞样细胞。

Objective To identify gastric cancer stem-like cells in SGC-7901 cell line. Methods SGC-7901 cells were cultured in the serum free media at a density of 1 000 cells/mL tO form spheres. Side Population （SP） was tested by flow cytometry to identify cancer stem-like cells. The in vivo NOD/SCID xenograft assay was done to test their tumor-forming capacity. Microarray experiment was used to search for differentially expressed genes between floating spheres and adherent cells, and the re- sults were further confirmed by real-time flurogenetic quantitative PCR （RFQ-PCR）. Results Only （8.00±0.54）% of SGC-7901 cells could form floating spheres in serum free media, which contained （12.34±1.01）% of SP cells. Tumor formation assay showed that sphere cells were more tumorigenic than adherent cells. Microarray experiment revealed that sphere cells highly expressed stem cell associat- ed proteins. ABCG2, HES1 and KRT153, which were verified by RFQ-PCR. Conclusions SGC-7901 cell line contains gastric cancer stem-like cells.