Impacts of ethanol on the epidermal growth factor receptor (EGFR) -calpain signaling and migration in breast cancer cells

李勇杰; 于庆龙; 潘际刚

doi:10.19401/j.cnki.1007-3639.2016.10.003

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Impacts of ethanol on the epidermal growth factor receptor (EGFR) -calpain signaling and migration in breast cancer cells

更新时间：2025-12-31

- Impacts of ethanol on the epidermal growth factor receptor (EGFR) -calpain signaling and migration in breast cancer cells
- China Oncology Vol. 26, Issue 10, Pages: 820-825(2016)
- 作者机构：
  
  贵州医科大学基础医学院生理学教研室,贵州,贵阳,550025
- 作者简介：
- 基金信息：
- DOI：10.19401/j.cnki.1007-3639.2016.10.003
  CLC：
- Published Online：17 November 2016，
  
  Published：17 November 2016
- 稿件说明：
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李勇杰,于庆龙,潘际刚,等. 酒精对乳腺癌细胞表皮生长因子受体-钙激活中性蛋白酶通路及细胞迁移的影响[J]. 中国癌症杂志, 2016, 26(10): 820-825.

李勇杰, 于庆龙, 潘际刚. Impacts of ethanol on the epidermal growth factor receptor (EGFR) -calpain signaling and migration in breast cancer cells[J]. China Oncology, 2016, 26(10): 820-825.
李勇杰,于庆龙,潘际刚,等. 酒精对乳腺癌细胞表皮生长因子受体-钙激活中性蛋白酶通路及细胞迁移的影响[J]. 中国癌症杂志, 2016, 26(10): 820-825. DOI： 10.19401/j.cnki.1007-3639.2016.10.003.

李勇杰, 于庆龙, 潘际刚. Impacts of ethanol on the epidermal growth factor receptor (EGFR) -calpain signaling and migration in breast cancer cells[J]. China Oncology, 2016, 26(10): 820-825. DOI： 10.19401/j.cnki.1007-3639.2016.10.003.

摘要

背景与目的：酒精(ethyl alcohol，EtOH)可促进乳腺癌的恶性演进，但其信号机制尚未完全明了。本研究通过观察EtOH对乳腺癌细胞钙激活中性蛋白酶(calcium-activated neutral protease，CANP)-周期蛋白E/局部黏着斑激酶(focal adhesion kinase，FAK)信号通路和细胞迁移的影响，以及表皮生长因子受体(epidermal growth factor receptor，EGFR)在其中的介导作用，探讨EtOH促进乳腺癌细胞迁移的信号机制。方法：以人乳腺肿瘤细胞系MCF-7为模型细胞(MCF-10A乳腺上皮细胞为对照)；采用蛋白［质］印迹法(Western blot)分析周期蛋白E和FAK蛋白剪切反应；通过伤口愈合实验观察细胞迁移效应；采用EGFR抑制剂(EGFR inhibitor，EGFR-I)或CANP抑制剂Calpeptin抑制EGFR或CANP活性，观察抑制剂对EtOH诱导CANP1大亚基、周期蛋白E/FAK蛋白剪切及细胞迁移的影响。结果：以EtOH(0.3%)处理模型细胞可观察到周期蛋白E和FAK出现明显蛋白剪切且该效应呈剂量和时间依赖性，还观察到EtOH刺激细胞迁移活动(+47.30%，P0.05)，但EtOH对MCF-10A细胞迁移无明显影响；以Calpeptin(10 μmol/L)预处理模型细胞，可见EtOH(0.3%)或EGF(10 ng/mL)诱导的周期蛋白E/FAK蛋白剪切效应受到明显抑制；EGFR-I(3 μmol/L)可显著抑制EtOH诱导的CANP1大亚基和周期蛋白E/FAK蛋白剪切及细胞迁移效应(-53.00%，P0.01)。结论：EtOH可通过EGFR激活乳腺癌细胞CANP信号活动并促进细胞迁移，EGFR-CANP通路参与介导EtOH与EGF之间的串话，提示EGFR-CANP信号通路中的关键分子可为防止乳腺癌转移的潜在药物靶点。

Abstract

Background and purpose: Ethanol has been reported to stimulate progression of breast cancer

yet the underlying mechanism is not fully understood. This study aimed to investigate effects of ethyl alcohol (EtOH) on the calcium-activated neutral protease (CANP)-cyclin E/focal adhesion kinase (FAK) signaling and cell migration in breast cancer cells

as well as the role of epidermal growth factor receptor (EGFR) in the EtOH-stimulated effects

in order to assess the signaling mechanism(s) underlying how EtOH enhances cancer progression. Methods: Human breast cancer cell line MCF-7 was employed as a model system

with MCF-10A mammary epithelial cells as control. In vitro wound healing assay was carried out to evaluate EtOH-induced cell migration. The effects of EtOH or epidermal growth factor on the proteolysis of cyclin E/FAK were detected by Western blot. EGFR inhibitor (EGFR-I) and a specific inhibitor for CANP

Calpeptin

were applied to pretreat cultured cells to explore their influences on the cell migration and cyclin E/FAK proteolysis triggered by EtOH. Results: Treatment of model cells with EtOH (0.3%) stimulated significant proteolysis of cyclin E/FAK in a dose-/time-dependent manner and increased migration (+47.30%

P0.05) in MCF-7 breast cancer cells

but had no significant effect on migration in MCF-10A cells. Pretreatment with Calpeptin (10 μmol/L) significantly reduced EtOH (0.3%)- or EGFR (10 ng/mL)-induced cyclin E/FAK truncation. EGFR-I (3 μmol/L) profoundly reduced EtOH-indcued CANP dependent proteolysis of CANP1 and cyclin E/FAK as well as cell migration (-53.00%

P0.01). Conclusion: EtOH significantly stimulates activation of CANP via EGFR pathway

resulting in proteolysis of cyclin E/FAK and migration in MCF-7 breast cancer cells

suggesting EGFR-CANP signaling to be a potential target for suppression of metastasis in breast cancer.

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