Invasive potential induced under long-term oxidative stress in mammary epithelial cells
Invasive potential induced under long-term oxidative stress in mammary epithelial cells
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DOI：
10.1158/0008-5472.can-04-1725
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发表时间：
2004-10-15
期刊：
影响因子：
11.2
通讯作者：
Nose, K
中科院分区：
文献类型：
--
作者：
Mori, K;Shibanuma, M;Nose, K
尽管长期以来人们一直在讨论慢性炎症与致癌之间的因果关系，但人们对这种关系的分子基础知之甚少。在本研究中，我们关注炎症条件下活性氧（ROS）及其信号导致上皮细胞癌变，发现用低剂量H2O2（0.2mmol/L）重复治疗2至4天会导致小鼠NMuMG乳腺上皮细胞从上皮细胞向成纤维细胞样细胞发生恶性转化的表型转变。使用 Northern blot 分析和定量逆转录 PCR 进行分析，表型转变包括细胞间接触的溶解、E-钙粘蛋白在细胞质中的重新分布以及一组整合素家族成员（整合素 α2、α6 和 β3）和基质金属蛋白酶（MMP；MMP-3、-10 和 -13）的上调。明胶酶谱显示明胶酶（包括 MMP-2 和 -9）的转录后激活。同时，p38 丝裂原激活蛋白激酶和细胞外信号调节激酶 1/2 被激活，这有助于 MMP-13 的诱导，谷胱甘肽 S 转移酶下拉分析显示小 GTP 酶 Rac1 的激活。令人惊讶的是，延长的氧化处理足以诱发所有上述事件。最重要的是，根据MMP的活性，暴露于氧化条件下的上皮细胞最终在底部含有正常成纤维细胞的基质胶侵袭室的重建模型系统中获得侵袭性，为支持ROS信号在上皮细胞恶性转化中的直接作用提供了第一个实质性证据。
Although the causal relationship between chronic inflammation and carcinogenesis has long been discussed, the molecular basis of the relation is poorly understood. In the present study, we focused on reactive oxygen species (ROS) and their signals under inflammatory conditions leading to the carcinogenesis of epithelial cells and found that repeated treatment with a low dose of H2O2 (0.2 mmol/L) for periods of 2 to 4 days caused a phenotypic conversion of mouse NMuMG mammary epithelial cells from epithelial to fibroblast-like as in malignant transformation. The phenotypic conversion included the dissolution of cell-cell contacts, redistribution of E-cadherin in the cytoplasm, and up-regulation of a set of integrin family members (integrin alpha2, alpha6, and beta3) and matrix metalloproteinases (MMPs; MMP-3, -10, and -13), as analyzed using Northern blot analysis and quantitative reverse transcription-PCR. Gelatin zymography indicated post-transcriptional activation of gelatinases, including MMP-2 and -9. In parallel, p38 mitogen-activated protein kinase and extracellular signal-regulated kinase 1/2 were activated, which contributed to the induction of MMP-13, and a glutathione S-transferase pull-down assay showed the activation of a small GTPase, Rac1. Surprisingly, the prolonged oxidative treatment was sufficient to induce all of the aforementioned events. Most importantly, depending on the MMP activities, the epithelial cells exposed to oxidative conditions eventually acquired invasiveness in a reconstituted model system with a Matrigel invasion chamber containing normal fibroblasts at the bottom, providing the first substantial evidence supporting the direct role of ROS signals in the malignant transformation of epithelial cells.