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Differentiation of immature enterocytes into enteroendocrine cells by Pdx-1 overexpression.

通过 Pdx-1 过表达将未成熟肠上皮细胞分化为肠内分泌细胞。

基本信息

批准号：
13670010
负责人：
FUJIMIYA Mineko
金额：
$ 2.3万
依托单位：
Shiga University of Medical Sgience
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670010/
关键词：
Intestinal Stem cell IEC6 cell Pdx-1 transcriptinal factor enteroendocrine cells Pancreatic B cells

项目摘要

The development of a variety of enteroendocrine cells of the gut is poorly understood. We tested whether immature intestinal stem cells were switched to multiple enteroendocrine hormone-producing cells by in vitro transfer of a homeobox gene. We transfected the pancreatic-duodenal homeobox 1 gene (Pdx1) into IEC-6 cells, an embryonic intestinal epithelial cell line derived from a normal rat, and selected the cells that overexpressed Pdx1 by 150-fold compared with control. The cells were examined for differentiation into enteroendocrine cells by immunocytochemical and electron microscopic analyses. Transfected cells cultured on micropore filters formed a trabecular network piled up on monolayer cells. These trabecular cells showed nuclear localization of Pdx1 protein and contained well-developed rough endoplasmic reticulum as well as many secretory granules of pleomorphic shape in the cytoplasm. Antibodies against chromogranin A, serotonin, cholecystokinin, gastrin, and somatostatin stained these secretory granules in the cytoplasm. Furthermore, immunofluorescence double staining analysis showed that different hormones were produced within a cell. These results provide the evidence that immature intestinal epithelial cells can differentiate into multiple hormone-producing enteroendocrine cells in response to overexpression of Pdx1.

对肠道各种肠内分泌细胞的发育知之甚少。我们测试了是否未成熟的肠干细胞转换为多个肠内分泌肿瘤产生细胞的同源异型盒基因的体外转移。我们将胰腺十二指肠同源盒1基因（Pdx 1）转染到IEC-6细胞中，IEC-6细胞是一种来自正常大鼠的胚胎肠上皮细胞系，并选择了与对照相比过表达Pdx 1 150倍的细胞。通过免疫细胞化学和电子显微镜分析检测细胞向肠内分泌细胞的分化。转染细胞在单层细胞上形成小梁网。这些小梁细胞显示Pdx 1蛋白的核定位，并含有发达的粗面内质网以及许多多形性的分泌颗粒在胞质中。抗嗜铬粒蛋白A、5-羟色胺、胆囊收缩素、胃泌素和生长抑素的抗体染色这些细胞质中的分泌颗粒。此外，免疫荧光双染色分析表明，不同的激素在一个细胞内产生。这些结果提供的证据表明，未成熟的肠上皮细胞可以分化成多个细胞产生肠内分泌细胞响应Pdx 1的过表达。