Moreno-Amador José Luis, Téllez Noèlia, Marin Sandra, Aloy-Reverté Caterina, Semino Carlos, Nacher Montserrat, Montanya Eduard
Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), Barcelona, Spain.
CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Madrid, Spain.
PLoS One. 2018 Jan 23;13(1):e0191104. doi: 10.1371/journal.pone.0191104. eCollection 2018.
β-cells undergo an epithelial to mesenchymal transition (EMT) when expanded in monolayer culture and give rise to highly proliferative mesenchymal cells that retain the potential to re-differentiate into insulin-producing cells.
To investigate whether EMT takes place in the endocrine non-β cells of human islets.
Human islets isolated from 12 multiorgan donors were dissociated into single cells, purified by magnetic cell sorting, and cultured in monolayer.
Co-expression of insulin and the mesenchymal marker vimentin was identified within the first passage (p1) and increased subsequently (insulin+vimentin+ 7.2±6% at p1; 43±15% at p4). The endocrine non-β-cells did also co-express vimentin (glucagon+vimentin+ 59±1.5% and 93±6%, somatostatin+vimentin+ 16±9.4% and 90±10% at p1 and p4 respectively; PP+vimentin+ 74±14% at p1; 88±12% at p2). The percentage of cells expressing only endocrine markers was progressively reduced (0.6±0.2% insulin+, 0.2±0.1% glucagon+, and 0.3±0.2% somatostatin+ cells at p4, and 0.7±0.3% PP+ cells at p2. Changes in gene expression were also indicated of EMT, with reduced expression of endocrine markers and the epithelial marker CDH-1 (p<0.01), and increased expression of mesenchymal markers (CDH-2, SNAI2, ZEB1, ZEB2, VIM, NT5E and ACTA2; p<0.05). Treatment with the EMT inhibitor A83-01 significantly reduced the percentage of co-expressing cells and preserved the expression of endocrine markers.
In adult human islets, all four endocrine islet cell types undergo EMT when islet cells are expanded in monolayer conditions. The presence of EMT in all islet endocrine cells could be relevant to design of strategies aiming to re-differentiate the expanded islet cells towards a β-cell phenotype.
β细胞在单层培养中扩增时会经历上皮-间质转化（EMT），并产生具有高增殖能力的间充质细胞，这些细胞保留了重新分化为胰岛素分泌细胞的潜力。
研究EMT是否发生在人胰岛的内分泌非β细胞中。
从12名多器官供体分离的人胰岛解离为单细胞，通过磁珠细胞分选纯化，并进行单层培养。
在第1代（p1）中鉴定出胰岛素与间充质标志物波形蛋白的共表达，随后增加（p1时胰岛素+波形蛋白+细胞占7.2±6%；p4时占43±15%）。内分泌非β细胞也共表达波形蛋白（p1和p4时，胰高血糖素+波形蛋白+细胞分别占59±1.5%和93±6%，生长抑素+波形蛋白+细胞分别占16±9.4%和90±10%；p1时胰多肽+波形蛋白+细胞占74±14%；p2时占88±12%）。仅表达内分泌标志物的细胞百分比逐渐降低（p4时胰岛素+细胞占0.6±0.2%，胰高血糖素+细胞占0.2±0.1%，生长抑素+细胞占0.3±0.2%；p2时胰多肽+细胞占0.7±0.3%）。基因表达变化也表明发生了EMT，内分泌标志物和上皮标志物CDH-1的表达降低（p<0.01），间充质标志物（CDH-2、SNAI2、ZEB1、ZEB2、VIM、NT5E和ACTA2）的表达增加（p<0.05）。用EMT抑制剂A83-01处理显著降低了共表达细胞的百分比，并保留了内分泌标志物的表达。
在成人胰岛中，当胰岛细胞在单层条件下扩增时，所有四种内分泌胰岛细胞类型都会发生EMT。所有胰岛内分泌细胞中EMT的存在可能与旨在将扩增的胰岛细胞重新分化为β细胞表型的策略设计相关。
