Endothelin-3 stimulates survival of goblet cells in organotypic cultures of fetal human colonic epithelium
Endothelin-3 stimulates survival of goblet cells in organotypic cultures of fetal human colonic epithelium
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DOI：
10.1152/ajpgi.90294.2008
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发表时间：
2008-12-01
影响因子：
4.5
通讯作者：
Herlyn, Meenhard
中科院分区：
文献类型：
--
作者：
Kalabis, Jiri;Li, Gang;Herlyn, Meenhard
正常人结肠上皮内的细胞经历生长、分化和死亡的动态循环。接种在具有包埋的结肠成纤维细胞的胶原凝胶上的人胎儿结肠上皮细胞的器官型培养系统允许结肠上皮细胞的延长培养（Kalabis J，Patterson MJ，Enders GM，Marian B，Iozzo RV，Rogler G，Gimotty PA，Herlyn M. FASEB J 17：1115-1117，2003）。在此，我们评估了内皮素-3（ET 3）和两种同源内皮素受体（ETRA，ETRB）对人结肠上皮细胞生长和存活的作用。由于腺病毒介导的基因转移，成纤维细胞持续产生ET 3。内皮素受体（ETRs）在上皮细胞中的存在和功能进行了评价[H-3]胸苷掺入使用原代上皮细胞在单一培养和免疫组化对人胎儿和成人石蜡包埋组织。在器官型培养中，ET 3增加杯状细胞的数量，但不增加肠内分泌细胞的数量。杯状细胞的增加是由于细胞存活和分化时间延长所致。抑制ETRA和ETRB显着减少杯状细胞的数量和上皮细胞的增殖，而肠内分泌细胞的数量保持不变。ET 3可诱导上皮细胞I kappa B和MAPK的活化，提示这些信号通路介导ET 3的促增殖和促存活活性。我们的研究结果表明，ET 3参与调节人类结肠上皮细胞的增殖和存活，特别是杯状细胞，并可能是结肠稳态的重要组成部分。
Cells within the normal human colonic epithelium undergo a dynamic cycle of growth, differentiation, and death. The organotypic culture system of human fetal colonic epithelial cells seeded on top of collagen gels with embedded colonic fibroblasts allowed prolonged culture of the colonic epithelial cells (Kalabis J, Patterson MJ, Enders GM, Marian B, Iozzo RV, Rogler G, Gimotty PA, Herlyn M. FASEB J 17: 1115-1117, 2003). Herein, we have evaluated the role of endothelin-3 (ET3) and both cognate endothelin receptors (ETRA, ETRB) for human colonic epithelial cell growth and survival. ET3 was produced continuously by the fibroblasts as a result of adenovirus-mediated gene transfer. The presence and function of the endothelin receptors (ETRs) in epithelial cells was evaluated by [H-3]thymidine incorporation using primary epithelial cells in monoculture and by immunohistochemistry on human fetal and adult paraffin-embedded tissues. In organotypic culture, ET3 increased the number of goblet cells but not of enteroendocrine cells. The increase in goblet cells was caused by prolonged cell survival and differentiation. The inhibition of both ETRA and ETRB significantly decreased the number of goblet cells and proliferation in epithelial cells, whereas the number of enteroendocrine cells remained unchanged. ET3 induced activation of I kappa B and MAPK in the epithelial cells, suggesting that these signaling pathways mediate its proproliferation and prosurvival activities. Our results demonstrate that ET3 is involved in regulating human colonic epithelial cell proliferation and survival, particularly for goblet cells, and may be an important component of colonic homeostasis.