过刊浏览

单功能烷化剂MNNG对人胃粘膜上皮GES-1细胞的损伤及其对Wnt/β-catenin信号通路的影响

严展鹏^1,2, 徐婷婷^1,2, 安振涛^1,2, 胡莹^1,2, 陈婉珍^1,2, 朱方石^1,2,*

¹南京中医药大学附属中西医结合医院，南京 210028；²江苏省中医药研究院，南京 210028

摘要

本文旨在从Wnt/β-catenin信号通路的角度探讨单功能烷化剂MNNG诱导人胃粘膜上皮GES-1细胞损伤的机制。用MNNG (2 × 10−5 mol/L)处理GES-1细胞24 h，处理后第3天用倒置显微镜对GES-1细胞形态进行观察，用MTT法检测GES-1细胞活力，用流式细胞术检测GES-1细胞凋亡和周期变化，用qPCR检测GES-1细胞中β-catenin、GSK-3β、c-Met和MMP7 mRNA表达情况，用Western blot检测β-catenin、GSK-3β、p-GSK-3β和c-Met蛋白表达情况。结果显示：MNNG能够明显损伤GES-1细胞，细胞形态变为不规则的长梭形；MNNG能够诱导GES-1细胞凋亡，明显阻滞细胞周期进程；MNNG能够上调β-catenin、GSK-3β、c-Met和MMP7 mRNA表达水平，并上调β-catenin、GSK-3β和p-GSK-3β蛋白表达水平。上述结果提示，MNNG对GES-1细胞的损伤可能与Wnt/β-catenin信号通路的激活相关，这为胃粘膜损伤的细胞模型研究提供了参考。

关键词： Wnt/β-catenin信号通路; 胃粘膜上皮细胞; 损伤机制

分类号：Q48;R573

Injury of human gastric epithelial GES-1 cells by MNNG and its effects on Wnt/β-catenin signaling pathway

YAN Zhan-Peng^1,2, XU Ting-Ting^1,2, AN Zhen-Tao^1,2, HU Ying^1,2, CHEN Wan-Zhen^1,2, ZHU Fang-Shi^1,2,*

¹Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing 210028, China；²Jiangsu Province Institute of Traditional Chinese Medicine, Nanjing 210028, China

Abstract

The aim of this study was to investigate the mechanisms of mono-functional alkylating agent MNNG to damage human gastric epithelial GES-1 cells and roles of Wnt/β-catenin signaling pathway in the process. The GES-1 cells were treated with MNNG (2 × 10−5 mol/L) for 24 h. The morphological changes of the GES-1 cells were observed under inverted microscope 2 d after treatment. The cell viability was measured by MTT assay. The apoptosis and cell cycle distribution of the GES-1 cells were analyzed by flow cytometry. The mRNA expressions of β-catenin, GSK-3β, c-Met and MMP7 in the GES-1 cells were detected by qPCR. The protein expressions of β-catenin, GSK-3β, p-GSK-3β and c-Met were determined by Western blot. The results showed that MNNG induced the injury of GES-1 cells and changed the normal cell morphology to irregular long spindle shape. MNNG induced the apoptosis of GES-1 cells and blocked the cell cycle progression obviously. MNNG up-regulated the mRNA expressions of β-catenin, GSK-3β, c-Met and MMP7, and increased the protein expressions of β-catenin, GSK-3β and p-GSK-3β. These results suggest that the damage of GES-1 cells induced by MNNG may be related to the activation of Wnt/β-catenin signaling pathway, which will provide the basis for the study of cell model of gastric mucosal cell injury.

Key words: Wnt/β-catenin signaling pathway; gastric mucosa epithelial cells; damage mechanisms

收稿日期：2017-11-17　　录用日期：2018-01-04

通讯作者：朱方石　　E-mail: zhufangshi@126.com

DOI: 10.13294/j.aps.2018.0042

引用本文：

严展鹏, 徐婷婷, 安振涛, 胡莹, 陈婉珍, 朱方石. 单功能烷化剂MNNG对人胃粘膜上皮GES-1细胞的损伤及其对Wnt/β-catenin信号通路的影响[J]. 生理学报 2018; 70 (3): 262-268.

YAN Zhan-Peng, XU Ting-Ting, AN Zhen-Tao, HU Ying, CHEN Wan-Zhen, ZHU Fang-Shi. Injury of human gastric epithelial GES-1 cells by MNNG and its effects on Wnt/β-catenin signaling pathway. Acta Physiol Sin 2018; 70 (3): 262-268 (in Chinese with English abstract).