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Pancreatic stellate cells exhibit adaptation to oxidative stress evoked by hypoxia
Biology of the Cell ( IF 2.4 ) Pub Date : 2020-07-12 , DOI: 10.1111/boc.202000020 Matias Estaras 1 , Salome Martinez-Morcillo 2 , Alfredo García 3 , Remigio Martinez 4 , Mario Estevez 5 , Marcos Perez-Lopez 2 , Maria P Miguez 2 , Miguel Fernandez-Bermejo 6 , Jose M Mateos 6 , Daniel Vara 6 , Gerardo Blanco 7 , Diego Lopez 7 , Vicente Roncero 8 , Gines M Salido 1 , Antonio Gonzalez 1
Biology of the Cell ( IF 2.4 ) Pub Date : 2020-07-12 , DOI: 10.1111/boc.202000020 Matias Estaras 1 , Salome Martinez-Morcillo 2 , Alfredo García 3 , Remigio Martinez 4 , Mario Estevez 5 , Marcos Perez-Lopez 2 , Maria P Miguez 2 , Miguel Fernandez-Bermejo 6 , Jose M Mateos 6 , Daniel Vara 6 , Gerardo Blanco 7 , Diego Lopez 7 , Vicente Roncero 8 , Gines M Salido 1 , Antonio Gonzalez 1
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Pancreatic stellate cells play a key role in the fibrosis that develops in diseases such as pancreatic cancer. In the growing tumour, a hypoxia condition develops under which cancer cells are able to proliferate. The growth of fibrotic tissue contributes to hypoxia. In this study, the effect of hypoxia (1% O2) on pancreatic stellate cells physiology was investigated. Changes in intracellular free‐Ca2+ concentration, mitochondrial free‐Ca2+ concentration and mitochondrial membrane potential were studied by fluorescence techniques. The status of enzymes responsible for the cellular oxidative state was analyzed by quantitative reverse transcription‐polymerase chain reaction, high‐performance liquid chromatography, spectrophotometric and fluorimetric methods and by Western blotting analysis. Cell viability and proliferation were studied by crystal violet test, 5‐bromo‐2‐deoxyuridine cell proliferation test and Western blotting analysis. Finally, cell migration was studied employing the wound healing assay.
中文翻译：
胰腺星状细胞表现出对缺氧引起的氧化应激的适应
胰腺星状细胞在胰腺癌等疾病发生的纤维化中起着关键作用。在生长的肿瘤中，会出现缺氧条件，在此条件下癌细胞能够增殖。纤维化组织的生长导致缺氧。在这项研究中，研究了缺氧 (1% O2) 对胰腺星状细胞生理学的影响。通过荧光技术研究细胞内游离Ca2+浓度、线粒体游离Ca2+浓度和线粒体膜电位的变化。通过定量逆转录聚合酶链反应、高效液相色谱、分光光度法和荧光法以及蛋白质印迹分析来分析负责细胞氧化状态的酶的状态。通过结晶紫试验、5-溴-2-脱氧尿苷细胞增殖试验和蛋白质印迹分析研究细胞活力和增殖。最后，使用伤口愈合试验研究了细胞迁移。
更新日期：2020-07-12
中文翻译：
胰腺星状细胞表现出对缺氧引起的氧化应激的适应
胰腺星状细胞在胰腺癌等疾病发生的纤维化中起着关键作用。在生长的肿瘤中，会出现缺氧条件，在此条件下癌细胞能够增殖。纤维化组织的生长导致缺氧。在这项研究中，研究了缺氧 (1% O2) 对胰腺星状细胞生理学的影响。通过荧光技术研究细胞内游离Ca2+浓度、线粒体游离Ca2+浓度和线粒体膜电位的变化。通过定量逆转录聚合酶链反应、高效液相色谱、分光光度法和荧光法以及蛋白质印迹分析来分析负责细胞氧化状态的酶的状态。通过结晶紫试验、5-溴-2-脱氧尿苷细胞增殖试验和蛋白质印迹分析研究细胞活力和增殖。最后，使用伤口愈合试验研究了细胞迁移。
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