转染胃动蛋白2基因抑制人胃癌MKN28细胞系增殖、迁移和侵袭

基础医学与临床 ›› 2018, Vol. 38 ›› Issue (3): 370-374.

转染胃动蛋白2基因抑制人胃癌MKN28细胞系增殖、迁移和侵袭

蔡志宏,曹怡静,刘姣,胡皓彬,谢鑫,许雄峰,凌晖

南华大学

收稿日期:2017-03-31 修回日期:2017-07-03 出版日期:2018-03-05 发布日期:2018-02-27
通讯作者: 凌晖 E-mail:nhdxlh@163.com
基金资助:
湖南省教育厅科学研究项目;湖南省十二五重点学科“基础医学”;肿瘤细胞与分子病理学湖南省重点实验室资助;湖南省(南华大学)研究生科研创新项目

GKN2 transfection inhibits proliferation, migration and invasion of gastric cancer cell line MKN28

Received:2017-03-31 Revised:2017-07-03 Online:2018-03-05 Published:2018-02-27

摘要/Abstract

摘要： 目的检测胃癌、癌旁组织和胃癌远端胃黏膜组织中胃动蛋白2（GKN2）的表达；分析GKN2转染人胃癌MKN28细胞系后对增殖、迁移和侵袭的影响。方法免疫组织化学技术检测胃癌、癌旁组织和胃癌远端胃黏膜中GKN2蛋白的表达；将GKN2基因真核表达载体（Xhol_GKN3SP-hGKN2-TEV-SBP_Xhol）转入人胃癌MKN28细胞，经G418挑选后获得稳转细胞株，Western blot确定GKN2在MKN28细胞中表达；MTT测定MKN28细胞增殖；Transwell迁移实验和侵袭实验检测细胞迁移和侵袭。结果 GKN2在胃癌组织中的表达少于胃癌远端胃黏膜和癌旁胃组织（P＜0.05）。GKN2转染人胃癌MKN28细胞后吸光度值（A值）显著低于未转染组和空载体组（P＜0.05）。与未转染组及空载体组对比，GKN2过表达致使迁移细胞数（P＜0.05）和侵袭细胞数均明显下调（P＜0.05）。结论 GKN2表达减低与胃癌的发生有关；GKN2过表达显然可阻抑人胃癌MKN28细胞的增殖、迁移和侵袭。

关键词: 胃肿瘤, 胃动蛋白2, 增殖, 迁移, 侵袭

Abstract: Objective GKN2(gastrokines 2) protein expression is detected in gastric cancer tissues, the distal gastric mucosa tissues and the adjacent normal gastric tissues; the effect of GKN2 overexpression is displayed on proliferation, migration and invasion in human gastric cancer cell line MKN28. Methods Immunohistochemistry was performed to observe GKN2 expression in gastric cancinoma tissues, adjacent gastric mucosa tissues and distal gastric mucosa tissues. Then, the GKN2 gene eukaryotic expression vector Xhol_GKN3SP-hGKN2-TEV-SBP_Xhol was transfected into human gastric cancinoma MKN28 cells. Western blot was used to verify the expression of GKN2 in MKN28 cells. Cell viability, migration and invasion ability were investigated by MTT, Transwell migration assay and Transwell invasion assay. Results GKN2 expression in gastric cancinoma tissues (6.67%,6/90) lowered in comparison with those in the adjacent normal gastric tissues (43.75%,21/48) or distal gastric mucosa tissues (83.36%,19/22) respectively (P < 0.05). The viability of MKN28 cells was significantly inhibited after transfected by GKN2 vs untransfected cells (P < 0.05). GKN2 overexpression decreased the cells’ number passing through both the membrane and matrigel, compared to those of untransfected cells(P < 0.05). Conclusions Downregulation of GKN2 protein expression is associated with the occurrence of gastric cancer; GKN2 overexpression can suppress the proliferation, migration and invasion of MKN28 cells.

Key words: Stomach Neoplasms, Gastrokine 2 , Proliferation, Migration, Invasion

中图分类号:

中图分类号：R735.2

蔡志宏曹怡静刘姣胡皓彬谢鑫许雄峰凌晖. 转染胃动蛋白2基因抑制人胃癌MKN28细胞系增殖、迁移和侵袭[J]. 基础医学与临床, 2018, 38(3): 370-374.

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