Effect of Golgi α-mannosidase II expression inhibition by shRNA interference on invasive potential of human gastric cancer cell and its mechanism

Zhang Lu; Yang Yaying; Huang Ziyang; Yi Yongfen

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Effect of Golgi α-mannosidase II expression inhibition by shRNA interference on invasive potential of human gastric cancer cell and its mechanism

更新时间：2023-12-18

- Effect of Golgi α-mannosidase II expression inhibition by shRNA interference on invasive potential of human gastric cancer cell and its mechanism
- Journal of Chongqing Medical University Vol. 38, Issue 3, Pages: 310-317(2014)
- 作者机构：
  
  1. 重庆医科大学基础医学院病理学教研室、分子医学与肿瘤研究中心,重庆,400016
  2. 重庆医科大学附属第一医院老年科,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2014，
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Zhang Lu, Yang Yaying, Huang Ziyang, et al. Effect of Golgi α-mannosidase II expression inhibition by shRNA interference on invasive potential of human gastric cancer cell and its mechanism. [J]. Journal of Chongqing Medical University 38(3):310-317(2014)
DOI：

Zhang Lu, Yang Yaying, Huang Ziyang, et al. Effect of Golgi α-mannosidase II expression inhibition by shRNA interference on invasive potential of human gastric cancer cell and its mechanism. [J]. Journal of Chongqing Medical University 38(3):310-317(2014) DOI：

摘要

目的：初步分析高尔基体α-甘露糖苷酶Ⅱ（Golgi alpha-mannosidase Ⅱ，GMⅡ）在低中不同分化人胃癌细胞系中的差异表达，以探讨GMⅡ在胃癌发生发展中的作用。方法：应用阳离子脂质体法将合成的靶向GMⅡ基因的pGPU6/GFP/Neo-GMⅡ-1406质粒载体，分别转染至低分化胃癌细胞MGC-803和中分化胃癌细胞SGC-7901中，然后荧光倒置显微镜观察并计算转染率，最后经过G418筛选出稳定转染的细胞株。应用RT-PCR和Western blot检测转染后GMⅡ和E-cadherin、α-catenin 的mRNA及蛋白表达变化情况，同时利用细胞划痕实验和Transwell体外侵袭实验来测定GMⅡ被沉默后两株细胞侵袭能力的变化。结果：GMⅡ短发卡RNA（short hairpin RNA，shRNA）转染胃癌MGC-803和SGC-7901细胞后GMⅡ的mRNA和蛋白表达明显抑制，而E-cadherin和α-catenin的2种因子检测结果明显升高（P＜0.05）；细胞划痕实验结果显示，转染重组质粒pGPU6/GFP/Neo-GMⅡ-1406组（GMⅡshRNA组）与对照组（空白对照组，阴性对照组）相比，GMⅡ被沉默后的胃癌细胞的侵袭能力降低（P＜0.05）；Transwell体外侵袭实验中GMⅡshRNA组胃癌细胞较空白对照组、阴性对照组细胞侵袭能力都明显降低，18 h穿过小室的细胞数分别为：（65±5）、（197±6）、（186±5），（72±4）、（178±4）、（184±5）个与对照组相比差别具有统计学意义（MＧＣ-803：P=0.000，SGC-7901：P=0.000）。结论：GMⅡ基因沉默后的胃癌细胞的侵袭能力下降，可能与上调E-cadherin、α-catenin的表达有关。

Abstract

Objective：To analyze the expression of Golgi alpha-mannosidase Ⅱ（GMⅡ） in different gastric cancer cell lines and to in－vestigate the role of GMⅡ in gastric cancer genesis. Methods：The pGPU6/GFP/Neo-GMⅡ-1406 gene was transfected into MGC-803 and SGC-7901 cells using cationic liposome assay. Rates of transfected positive cells were observed and calculated under invert microscope. Gene and protein expression of GMⅡ，E-cadherin and α-catenin were tested using RT-PCR and Western blot respec－tively. Invasive abilities of cancer cells were analyzed by cell scratch and Transwell assay. Results：Protein expression of GMⅡ was significantly inhibited and expression of E-cadherin and α-catenin in MGC-803 and SGC-7901 cells was up-regulated compared with those of untransfected cells（P＜0.05）. Cell scratch assay showed that the invasive abilities of cells transfected with GMⅡ gene were weaker than those of control group cells（P＜0.05）. Transwell invasive assay demonstrated that the invasive abilities of cells trans－fected with GMⅡ gene were weaker than those of shNC groups. Numbers of Transwell cells were 65±5，197±6，186±5，72±4，178±4，184±5 in three groups during 18 h，with statistical differences（P＜0.05）. Conclusion：GMⅡ gene silencing contributes to decreased invasive ability of gastric cancer cell lines，which is involved with up-regulation of E-cadherin and α-catenin expression.

关键词

高尔基体α-甘露糖苷酶Ⅱ短发卡RNA胃癌侵袭

Keywords

Golgi alpha-mannosidase Ⅱshort hairpin RNAgastric cancerinvasion

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