转染 GKN2 基因对 CD44(+)SGC7901胃癌干细胞的抑制作用
Inhibitory Effect of GKN2 Gene Transfection on CD44(+)SGC7901 Gastric Cancer Stem Cells
胡皓彬 1凌晖 1蔡志宏 1肖萍 1李艳春1
作者信息
- 1. 湖南省人民医院/湖南师范大学附属第一医院病理科,湖南 长沙 410002
- 折叠
摘要
目的 研究胃动蛋白2(GKN2)高表达对CD44(+)SGC7901胃癌干细胞(GCSCs)的增殖、迁移、侵袭的影响.方法 采用免疫磁珠分选技术(MACS)从人胃癌细胞SGC7901中分选CD44(+)SGC7901 GCSCs,免疫荧光测试分选效率;将GKN2高表达腺病毒载体转染入GCSCs,Western Blot验证转染后GKN2的表达情况;CCK8、Transwell迁移侵袭实验观察GKN2高表达对人GCSCs增殖、迁移和侵袭的影响.结果 免疫荧光结果显示,CD44(+)SGC7901 GCSCs经免疫磁珠分选后数目大幅增加;CCK8增殖实验显示,GKN2转染组在450 nm处的OD值均低于未转染组及空载体组(P＜0.05);Transwell迁移实验结果显示,GKN2高表达组迁移及侵袭至下室的细胞数均低于未转染组及空载体组(P＜0.05).结论 GKN2高表达能抑制CD44(+)SGC7901 GCSCs的增殖,且能抑制其迁移及侵袭能力.
Abstract
Objective To investigate the effect of high expression of GKN2 on the proliferation,migration and invasion of CD44(+)SGC7901 gastric cancer stem cells(GCSCs).Methods CD44(+)SGC7901 GCSCs were isolated from human gastric cancer cell line SGC7901 by magnet-activated cell sorting(MACS),and the sorting efficiency was tested by immunofluorescence.The GKN2 high expression adenovirus vector was transfected into GCSCs,and the expression of GKN2 after transfection was verified by Western Blot.The effects of high expression of GKN2 on the proliferation,migration and invasion of human GCSCs were observed by CCK8 and Transwell migration and invasion experiments.Results Immunofluorescence results showed that the number of CD44(+)SGC7901 GCSCs increased significantly after magnet-activated cell sorting.CCK8 proliferation assay showed that the OD value of GKN2 transfection group at 450 nm was lower than that of non-transfection group and empty vector group(P＜0.05).Transwell migration assay showed that the number of cells migrating and invading to the lower chamber in the GKN2 high expression group was lower than that in the non-transfection group and the empty vector group(P＜0.05).Conclusion High expression of GKN2 can inhibit the proliferation,migration and invasion of CD44(+)SGC7901 GCSCs.
关键词
胃癌/胃癌干细胞/GKN2Key words
Gastric cancer/Gastric cancer stem cells/GKN2引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据