摘要
目的 探讨半乳糖凝集素3（Gal3）对食管癌Eca109细胞增殖和转移的影响.方法 构建合成Gal3过表达慢病毒质粒转染食管癌Eca109细胞,应用倒置显微镜观察Gal3的过表达质粒转染食管癌细胞后荧光表达;CCK-8法检测转染前后细胞增殖能力的变化;用流式细胞术检测转染组细胞和未转染组细胞凋亡率.Transwell方法检测转染前后细胞迁移能力变化;用Western印迹检测Gal3转染前后在食管癌中表达水平变化.结果 Western印迹结果显示Gal3在食管癌细胞中表达,在Eca109/Gal3组中表达水平明显升高（t=14.33,P=0.013;t=10.28,P=0.037）.CCK-8法检测发现转染后细胞增殖能力明显升高（t=-17.277,P＜0.05;t=-13.4,P＜0.05）,流式细胞仪以Annexin-V/7-AAD双标法显示Eca109/Gal3组凋亡率明显下降（t=3.053,P＜0.05;=5.446,P＜0.05）.Transwell实验结果,Eca109/Gal3组细胞转移率高于未转染组（t=3.465,P＜0.05;t=3.252,P＜0.05）.结论 Gal3在食管癌细胞中有表达,并且其过表达能显著增强食管癌细胞的增殖、侵袭及迁移能力,明显抑制细胞的凋亡,深入研究Gal3可能为食管癌的治疗提供一种新的靶点.
Objective To investigate galectin3 on proliferation and migration of esophageal cancer Eca109 cells.Methods A lentiviral vector for over-expression of RNA targeting galectin3 was designed to transfect Eca109 cancer cells following plasmid-mediated transfection manual （Eca109/Gal3 cells）.Inverted fluorescence microscope was used to observe the expression of EGFP.The proliferation of Eca109 cells was measured by cell counting Kit-8 assay.Eca109 cells apoptosis was determined by Annexin-V/7-AAD doublestaining.The migration capacity of Eca109 cells was determined in transwell assays.Western blot analysis was used to measure the expression of galectin3 protein.Results Galectin-3 expression was detected in Eca109 cells,with the Galectin3 expression in Eca109/Gal3 cells much more than non-transfected cells （t =14.33,P ＜ 0.05 ; t =10.28,P =0.037）.Compared with non-transfected Eca109 cells,proliferation increased significantly in Eca109/Gal3 cells （t =-17.277,P ＜ 0.05 ; t =-13.4,P ＜ 0.05）.Galectin3 evidently decreased in Eca109 cell apoptosis （t =3.053,P ＜ 0.05 ; t =5.446,P ＜ 0.05）.Transwell migration assay showed that a greater number of Eca109/Gal3 cells crossed the artificial basement membrane compared with non-transfected Eca109 cells and negative control Eca109 cells （t =3.465,P ＜ 0.05; t =3.252,P ＜ 0.05）.Conclusion Galectin3 expression is detected in transfected esophageal cancer Eca109 cells,whose overexpression can result in enhanced proliferation,migration,invasion as well as reduced apoptosis.These data indicate that in-depth research of galectin-3 may prove to be a potential molecular target for the treatment of esophageal cancer.
出处
《国际肿瘤学杂志》
CAS
2014年第5期375-379,共5页
Journal of International Oncology
基金
山东省自然科学基金（ZR2012HM095）
