Periostin调控AKT信号通路激活胃癌细胞的上皮间质转化

金良玉; 耿志军; 张小凤; 宋雪; 李静

doi:10.13898/j.cnki.issn.1000-2200.2023.04.001

Periostin调控AKT信号通路激活胃癌细胞的上皮间质转化

Periostin regulates AKT signaling pathway to activate epithelial-mesenchymal transition of gastric cancer cells

摘要

摘要:
目的探讨Periostin (POSTN)调控AKT信号通路激活胃癌细胞的上皮间质转化(EMT)的机制。
方法选取胃癌病人的癌旁、原发灶、转移灶以及不同临床分期的原发灶胃癌组织标本, 采用POSTN与CD44共同免疫荧光染色。选择人胃癌细胞MGC 803细胞, 并分别过表达和敲除POSTN基因, 分别添加AKT抑制剂LY294002进行孵育培养, 采用RT-qPCR法检测各组细胞的POSTN、CD44和EMT相关基因的表达水平; 采用蛋白印迹技术检测各组细胞的POSTN、CD44和EMT相关蛋白的表达水平; 采用CCK-8检测各组细胞的增殖水平; 采用Transwell实验检测细胞迁移和侵袭能力。
结果胃癌病人的转移灶中POSTN表达高于原发灶, 且原发灶中POSTN表达高于癌旁组织。与MGC 803细胞相比, 过表达POSTN基因的胃癌细胞中POSTN、CD44、α-sma、snail、slug、Vimentin表达升高和E-Cadherin表达降低以及增殖活性、迁移和侵袭能力升高; 敲除POSTN基因的胃癌细胞中CD44、α-sma、snail、slug、Vimentin表达降低以及E-Cadherin表达升高以及增殖活性、迁移和侵袭能力降低; 添加LY294002会显著地改善胃癌细胞中过表达POSTN基因引起的α-sma、snail、slug、Vimentin、E-Cadherin表达变化和增殖活性及迁移与侵袭能力的改变(P < 0.01)。
结论胃癌原发灶和转移灶中POSTN表达上调且临床分期越晚的原发灶中POSTN表达越高; POSTN通过调控AKT信号通路激活胃癌细胞的EMT, 进而促进胃癌细胞的增殖、迁移和侵袭。

Abstract:
ObjectiveTo investigate the mechanism of Periostin (POSTN) regulating AKT signaling pathway to activate the epithelial-mesenchymal transition (EMT) of gastric cancer cells.
MethodsPericancerous, cancerous, metastases and primary tumor tissues of patients with gastric cancer at different pathological stage were selected for immunofluorescence staining with POSTN and CD44.POSTN gene was overexpressed or knocked out in human gastric cancer cells MGC 803, and AKT inhibitor LY294002 was added for culture.The expression levels of POSTN, CD44 and EMT-related genes in gastric cancer cells were detected using RT-qPCR and Western blotting.The proliferation level of gastric cancer cells was detected using CCK-8.Transwell assays were used to detect the migration and invasion of cells.
ResultsThe expression of POSTN in metastasis was higher than that of primary tumor, and the expression of POSTN in primary tumor was higher than that of pericancerous tissue.Compared with normal MGC 803 cells, the expressions of POSTN, CD44, α-sma, snail, slug, Vimentin were increased and E-Cadherin was reduced in gastric cancer cells overexpressing POSTN, while POSTN knockout inhibited the expressions of CD44, α-sma, snail, slug, Vimentin and increased the expressions of E-Cadherin.The overexpression of POSTN promoted the metastasis and invasion of gastric cancer cells, while the reduction of POSTN inhibited the metastasis and invasion of gastric cancer cells.The addition of LY294002 significantly improved the expression changes of α-sma, snail, slug, Vimentin, E-Cadherin, proliferation activity, migration and invasion ability caused by overexpression of POSTN gene in gastric cancer cells(P < 0.01).
ConclusionsThe expression of POSTN in the cancerous and metastatic of gastric cancer is up-regulated, and the later the clinical stage, the higher the expression of POSTN in the cancerous; POSTN activates the EMT of gastric cancer cells by regulating the AKT signaling pathway, thereby promoting the proliferation, migration and invasion of gastric cancer cells.

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