A Preliminary Study of Side Population Cells in Human Gastric Cancer Cell Line HGC-27
A Preliminary Study of Side Population Cells in Human Gastric Cancer Cell Line HGC-27
复制标题
人胃癌细胞系HGC-27侧群细胞的初步研究
DOI：
10.12659/aot.892197
复制
发表时间：
2015-03-16
影响因子：
1.1
通讯作者：
Zhang, Xueli
中科院分区：
文献类型：
--
作者：
Gao, Ganglong;Sun, Zhenliang;Zhang, Xueli
背景资料：胃癌干细胞样侧群（SP）细胞在胃癌的多种细胞系中已被鉴定和表征，其可能负责肿瘤复发、转移和对癌症治疗的抗性。然而，从人胃癌细胞系HGC-27中分离SP细胞尚未见报道。本研究旨在分析HGC-27细胞系中SP细胞的比例，区分SP与非侧群（non-side population，NSP）细胞，并确定SP细胞是否具有干细胞的某些生物学特性。通过胃形成试验和定量真实的时间PCR检测SP和NSP细胞之间的增殖和干细胞相关基因表达谱（CD 133、CD 44、OCT-4、MDR 1、EpCAM和ABCG 2）的差异（3）。结果：（1）SP细胞占HGC-27细胞的0.1-1.0%，经维拉帕米抑制后，SP细胞占HGC-27细胞的比例降至0%;使用流式细胞术，我们分选了7.5x10（5）个SP细胞，并且大多数HGC-27细胞是NSP细胞（2）。胃形成实验和MTT法显示SP和NSP细胞的增殖能力有显著性差异。基因表达分析显示，基因的表达在SP细胞中显著更高（3）。结论：HGC-27细胞系SP细胞具有一定的肿瘤干细胞特性，可用于胃癌发病机制的研究，为寻找新的治疗靶点奠定基础。
Background: Cancer stem cell-like side population (SP) cells, which may be responsible for recurrence, tumor metastasis, and resistance to cancer therapy, have been identified and characterized in several types of cell lines from gastric cancer. However, there is no report on isolation of SP cells from human gastric cancer cell line HGC-27. This study aims to analyze the proportion of SP cells in HGC-27 cell line, differentiate SP from non-side population (NSP) cells, and determine whether the SP cells have certain biological properties of stem cells.Material/Methods: (1) HGC-27 suspension was prepared and stained with Hoechst33342 and PI for flow cytometric isolation of SP (2). Differences in proliferation and stemness-related gene expression profiles (CD133, CD44, OCT-4, MDR1, EpCAM, and ABCG2) between SP and NSP cells were detected by gastric formation assay and quantitative real- time PCR (3). Oncogenicity of SP and NSP cells was determined in nude mice in vivo.Results: (1) SP cells accounted for 0.1-1.0% of HGC-27 cells, and decreased to 0% after verapamil inhibition. Using flow cytometry, we sorted 7.5x10(5) SP cells and most HGC-27 cells were NSP cells (2). Gastric formation assay and MTT demonstrated that there was a significant difference in proliferation between SP and NSP cells. Gene expression analysis showed that the expression of genes was significantly higher in SP cells (3). The oncogenicity experiment in nude mice revealed that 105 SP cells were able to form tumors, which demonstrated higher tumorigenicity than non-SP cells.Conclusions: These results collectively suggested that SP cells from HGC-27 cell line have some cancer stem cell properties and could be used for studying the pathogenesis of gastric cancer, which may contribute to discovery of novel therapeutic targets.