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作 者:蔡志宏 曹怡静 刘姣[1] 胡皓彬 谢鑫 许雄峰[1] 凌晖[1]
机构地区:[1]南华大学医学院肿瘤研究所,湖南衡阳421001 [2]南华大学医学院,湖南衡阳421001
出 处:《基础医学与临床》2018年第3期370-374,共5页Basic and Clinical Medicine
基 金:湖南省教育厅科学研究项目(16C1398);湖南省十二五重点学科“基础医学”资助项目[2011(76)];肿瘤细胞与分子病理学湖南省重点实验室资助项目(2016TP1015);湖南省(南华大学)研究生科研创新项目(2016XCX37)
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-h GKN2-TEV-SBP_Xhol)转入人胃癌MKN28细胞,经G418筛选后获得稳转细胞株,Western blot确定GKN2在MKN28细胞中表达;MTT法测定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相比,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。Objective Gastrokines 2( GKN2) 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_GKN3 SP-h GKN2-TEV-SBP_Xhol was transfected into human gastric cancinoma MKN28 cells. Western blot was used to evaluate 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 is lower in comparison with those in the adjacent normal gastric tissues or distal gastric mucosa tissues 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.
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