上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

MACC-1通过HGF/c-Met信号通路调节上皮间质转化及其对胃癌细胞迁移和侵袭能力的影响

祝倩倩1，单海霞2，朱正秋2   

  1. 徐州医学院 1.研究生学院， 2.附属医院肿瘤科， 徐州 221002
  • 出版日期:2014-09-28 发布日期:2014-09-26
  • 通讯作者: 朱正秋, 电子信箱: js82880999@126.com。
  • 作者简介:祝倩倩（1988—）， 女， 硕士; 电子信箱: 15050829235@163.com。
  • 基金资助:

    江苏省卫生厅基金（H201323）

Regulation of epithelial-mesenchymal transition by MACC1 via the HGF/c-Met signaling pathway and its effects on ability of migration and invasion of gastric carcinoma cells

ZHU Qian-qian1, SHAN Hai-xia2, ZHU Zheng-qiu2   

  1. 1.School of Postgraduate, Xuzhou Medical College, Xuzhou 221002， China； 2.Department of Oncology, the Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China
  • Online:2014-09-28 Published:2014-09-26
  • Supported by:

    Jiangsu Province Department of Health Project, H201323

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摘要：

目的 探讨结肠癌转移相关基因-1 (MACC1)通过调控HGF/c-Met信号通路对上皮间质转化（EMT）的调节作用，以及对胃癌细胞迁移和侵袭能力的影响。方法 构建针对MACC1、c-Met的shRNA质粒载体（MACC1-shRNA、c-Met-shRNA）和阴性对照质粒（shNC），脂质体法转染人胃癌MKN28细胞株。采用Western blotting及Real-Time PCR技术检测转染前后MACC1、c-Met、EMT相关标志物（E-cadherin、N-cadherin）蛋白和mRNA表达变化。通过伤口愈合实验、Transwell侵袭实验检测MKN28细胞迁移和侵袭能力的变化。结果 与空白对照组相比，MACC1-shRNA组MACC1、c-Met蛋白及mRNA表达均明显下调（P<0.01），E-cadherin蛋白及mRNA表达水平均显著上调（P<0.01），N-cadherin蛋白及mRNA表达水平则显著下调（P<0.01）；c-Met-shRNA组MACC1蛋白和mRNA表达无明显变化（P>0.05），EMT相关标志物表达水平与MACC1-shRNA组的变化一致。伤口愈合实验和Transwell侵袭实验结果显示：与空白对照组相比，MACC1-shRNA组和c-Met-shRNA组MKN28细胞的迁移及侵袭能力均明显被抑制（P<0.01）。结论 MACC1可能通过调控HGF/c-Met信号通路调节EMT过程，增强胃癌细胞的迁移和侵袭能力。

关键词: 胃癌, 结肠癌转移相关基因-1, HGF/c-Met信号通路, 上皮间质转化, 迁移, 侵袭

Abstract:

Objective To investigate the regulatory effect of metastasis-associated gene in colon cancer-1 (MACC1) on the epithelial-mesenchymal transition (EMT) by regulating the HGF/c-Met signaling pathway and the effects of MACC1 on the migration and invasion of gastric carcinoma cells. Methods Short hairpin RNA (shRNA) plasmid vector that targeted MACC1 and c-Met (MACC1-shRNA and c-Met-shRNA) and negative control plasmid (shNC) were constructed and transfected into the human gastric carcinoma MKN28 cell line by the lipofectamine technique. The variations of expressions of proteins and mRNA of markers (E-cadherin and N-cadherin) related to MACC1, c-Met, and EMT before and after the transfection were detected by the Western blotting and Real-Time PCR. The variations of the ability of migration and invasion of MKN28 cells were detected by the wound-healing and Transwell invasion tests. Results Compared to the blank control group, the expressions of proteins and mRNA of MACC1 and c-Met of the MACC1-shRNA group were significantly down-regulated (P<0.01); the expressions of protein and mRNA of E-cadherin were significantly up-regulated (P<0.01); and the expressions of protein and mRNA of N-cadherin were significantly down-regulated (P<0.01). The expressions of protein and mRNA of MACC1 of the c-Met-shRNA group did not change significantly (P>0.05) and the variations of expressions of EMT related markers were consistent with those of the MACC1-shRNA group. The results of wound-healing and Transwell invasion tests showed that compared to the blank control group, the ability of migration and invasion of MKN28 cells of the MACC1-shRNA group and c-Met-shRNA group was significantly inhibited (P<0.01). Conclusion MACC1 may regulate the EMT process by regulating the HGF/c-Met signaling pathway and enhance the ability of migration and invasion of gastric carcinoma cells.

Key words: gastric carcinoma, metastasis-associated in colon cancer-1, HGF/c-Met signaling pathway, epithelial-mesenchymal transition, migration, invasion