Galán-Cobo Ana, Ramírez-Lorca Reposo, Serna Ana, Echevarría Miriam
Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla (Departamento de Fisiología Médica y Biofísica), Seville, Spain.
Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla (Departamento de Fisiología Médica y Biofísica), Seville, Spain; Centro de Investigación Biomédica en Red sobre Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Madrid, Spain.
PLoS One. 2015 Sep 14;10(9):e0137692. doi: 10.1371/journal.pone.0137692. eCollection 2015.
Abnormal AQP3 overexpression in tumor cells of different origins has been reported and a role for this enhanced AQP3 expression in cell proliferation and tumor processess has been indicated. To further understand the role AQP3 plays in cell proliferation we explore the effect that stable over expression of AQP3 produces over the proliferation rate and cell cycle of mammalian cells. The cell cycle was analyzed by flow cytometry with propidium iodide (PI) and the cell proliferation rate measured through cell counting and BrdU staining. Cells with overexpression of AQP3 (AQP3-o) showed higher proliferation rate and larger percentage of cells in phases S and G2/M, than wild type cells (wt). Evaluation of the cell response against arresting the cell cycle with Nocodazole showed that AQP3-o exhibited a less modified cell cycle pattern and lower Annexin V specific staining than wt, consistently with a higher resistance to apoptosis of AQP3-overexpressing cells. The cell volume and complexity were also larger in AQP3-o compared to wt cells. After transcriptomic analysis, RT-qPCR was performed to highlight key molecules implicated in cell proliferation which expression may be altered by overexpression of AQP3 and the comparative analysis between both type of cells showed significant changes in the expression of Zeb2, Jun, JunB, NF-kβ, Cxcl9, Cxcl10, TNF, and TNF receptors. We conclude that the role of AQP3 in cell proliferation seems to be connected to increments in the cell cycle turnover and changes in the expression levels of relevant genes for this process. Larger expression of AQP3 may confer to the cell a more tumor like phenotype and contributes to explain the presence of this protein in many different tumors.
据报道，不同来源的肿瘤细胞中存在异常的水通道蛋白3（AQP3）过表达，并且这种增强的AQP3表达在细胞增殖和肿瘤进程中发挥了作用。为了进一步了解AQP3在细胞增殖中的作用，我们探讨了AQP3稳定过表达对哺乳动物细胞增殖率和细胞周期的影响。通过碘化丙啶（PI）流式细胞术分析细胞周期，并通过细胞计数和BrdU染色测量细胞增殖率。与野生型细胞（wt）相比，过表达AQP3的细胞（AQP3-o）显示出更高的增殖率以及S期和G2/M期细胞的更大比例。用诺考达唑阻滞细胞周期后对细胞反应的评估表明，与wt相比，AQP3-o表现出较少改变的细胞周期模式和较低的膜联蛋白V特异性染色，这与AQP3过表达细胞对凋亡的更高抗性一致。与wt细胞相比，AQP3-o中的细胞体积和复杂性也更大。转录组分析后，进行逆转录定量聚合酶链反应（RT-qPCR）以突出参与细胞增殖的关键分子，其表达可能因AQP3过表达而改变，两种类型细胞之间的比较分析显示锌指E盒结合蛋白2（Zeb2）、Jun、JunB、核因子κB（NF-kβ）、CXC趋化因子配体9（Cxcl9）、CXC趋化因子配体10（Cxcl10）、肿瘤坏死因子（TNF）和TNF受体的表达有显著变化。我们得出结论，AQP3在细胞增殖中的作用似乎与细胞周期周转率的增加以及该过程相关基因表达水平的变化有关。AQP3的更大表达可能赋予细胞更像肿瘤的表型，并有助于解释该蛋白在许多不同肿瘤中的存在。
