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miR-208 诱导的胰腺癌上皮间质转化促进细胞转移和侵袭。

miR-208-induced epithelial to mesenchymal transition of pancreatic cancer cells promotes cell metastasis and invasion.

机构信息

Department of Pancreatic Surgery, Changhai Hospital, Second Military Medical University, No. 168 Changhai Road, Shanghai City, 200433, People's Republic of China.

出版信息

Cell Biochem Biophys. 2014 Jun;69(2):341-6. doi: 10.1007/s12013-013-9805-3.

Abstract

The aim of this study was to investigate the role of miR-208 in the invasion and metastasis of pancreatic cancer cells and the underlying molecular mechanism. miR-208 mimic, miR-208 inhibitor and NC were transfected into pancreatic cancer cell line Bxpc3 using liposome. Transwell invasion and scratch assays were used to test cell migratory and invasive abilities. Western blotting and quantitative PCR methods were used to detect E-cadherin, fibronectin and vimentin protein and mRNA expression in pancreatic cancer cell line BxPC3 after transfection by miR-208 mimic, miR-208 inhibitor and NC. Transwell invasion and scratch assays showed that after overexpressing miR-208, pancreatic cancer cell line BxPC3 exhibited enhanced in vitro migratory and invasive abilities, while after downregulating miR-208 expression, cell migratory and invasive abilities were decreased. Western blotting and quantitative PCR showed that after overexpressing miR-208, expression of E-cadherin, an epithelial cell marker, was decreased and expression of fibronectin and vimentin, interstitial cell markers, was increased in pancreatic cancer cell line BxPC3; however, after inhibiting miR-208, increased E-cadherin expression and decreased fibronectin and vimentin expression were observed in pancreatic cancer cell line BxPC3. After overexpressing miR-208, p-AKT and p-GSK-3β expression was altered by activating AKT/GSK-3β/snail signaling pathway. miR-208 induces epithelial to mesenchymal transition of pancreatic cancer cell line BxPC3 by activating AKT/GSK-3β/snail signaling pathway and thereby promotes cell metastasis and invasion.

摘要

本研究旨在探讨 miR-208 在胰腺癌细胞侵袭转移中的作用及其潜在的分子机制。采用脂质体将 miR-208 模拟物、miR-208 抑制剂和对照物(NC)转染至胰腺癌细胞系 Bxpc3 中。采用 Transwell 侵袭和划痕实验检测细胞迁移和侵袭能力。采用 Western blot 和 qPCR 方法检测 miR-208 模拟物、miR-208 抑制剂和 NC 转染后胰腺癌细胞系 Bxpc3 中 E-钙黏蛋白、纤连蛋白和波形蛋白的蛋白和 mRNA 表达。Transwell 侵袭和划痕实验表明,过表达 miR-208 后,胰腺癌细胞系 Bxpc3 体外迁移和侵袭能力增强,下调 miR-208 表达后,细胞迁移和侵袭能力降低。Western blot 和 qPCR 结果显示,过表达 miR-208 后,胰腺癌细胞系 Bxpc3 中上皮细胞标志物 E-钙黏蛋白的表达降低,间质细胞标志物纤连蛋白和波形蛋白的表达增加;而抑制 miR-208 后,胰腺癌细胞系 Bxpc3 中 E-钙黏蛋白表达增加,纤连蛋白和波形蛋白表达减少。过表达 miR-208 后,通过激活 AKT/GSK-3β/snail 信号通路改变了 p-AKT 和 p-GSK-3β 的表达。miR-208 通过激活 AKT/GSK-3β/snail 信号通路诱导胰腺癌细胞系 Bxpc3 的上皮间质转化,从而促进细胞转移和侵袭。

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