国际肿瘤学杂志››2014,Vol. 41››Issue (5): 375-379.doi:10.3760/cma.j.issn.1673-422X.2014.05.017

• 论著 •上一篇下一篇

半乳糖凝集素3对食管癌Eca109细胞增殖和转移的影响

梁宁,谢健,乔丽丽,张建东

  1. 261061潍坊医学院研究生院(梁宁);山东大学医学院研究生院(谢健、乔丽丽);山东大学附属千佛山医院肿瘤放疗科(张建东)
  • 收稿日期:2013-12-05修回日期:2014-02-25出版日期:2014-05-08发布日期:2014-04-28
  • 通讯作者:张建东 E-mail:zhangjd165@sina.com

Effect of galectin3 on proliferation and migration of esophageal cancer Eca109 cells

Liang Ning, Xie Jian, Qiao Lili, Zhang Jiandong

  1. Graduate School, Weifang Medical College, Weifang 261061, China
  • Received:2013-12-05Revised:2014-02-25Online:2014-05-08Published:2014-04-28
  • Contact:Zhang Jiandong E-mail:zhangjd165@sina.com

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;t=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。

关键词:食管肿瘤,半乳糖凝集素3,RNA过表达,生物学功能

Abstract: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 doublestaining. The migration capacity of Eca109 cells was determined in transwell assays. Western blot analysis was used to measure the expression of galectin3 protein. ResultsGalectin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 overexpression 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.

Key words:Esophageal neoplasms,Galectin 3,RNA over-expression,Biological function