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The effects of folate deficiency on on E-cadherin and beta-catenin in colon epithelial cells.

机译：叶酸缺乏对结肠上皮细胞中E-钙粘蛋白和β-连环蛋白的影响。

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摘要

Folate deficiency has been linked to multiple health problems including neural tube defects and cancer, especially colorectal cancer. Folate is an essential dietary vitamin that is required for DNA synthesis and cellular methylation reactions. The mechanistic links between these associations is not completely understood. Folate deficiency has been shown to affect genes associated with the Wnt pathway, including beta-catenin. E-cadherin is an important regulator of Wnt signaling through controlling the level of free beta-catenin. It has also been shown that E-cadherin is affected by folate deficiency. The purpose of this study was to determine if folate deficiency altered the levels of E-cadherin protein and RNA, beta-catenin protein, and E-cadherin and beta-catenin cellular localization in two colon epithelial cell lines. Caco-2 and FHC cells were grown for 15 and 30 days in folate sufficient and deficient medium. Cellular growth rates for both cell lines were reduced in folate deficient cells, although the cells continued to grow. Cell cycle phase distribution showed increases in S phase and G2/M phase in Caco-2 cells, whereas there was no change in FHC cells. E-cadherin and beta-catenin protein levels decreased slightly in folate deficient Caco-2 cells at days 15 and 30. This was associated with a decrease in E-cadherin transcripts at day 30. FHC cells showed similar results at day 15, but a small increase in E-cadherin protein at day 30 and no change in beta-catenin. Immunofluorescent staining of Caco-2 cells showed diffuse cytoplasmic staining of E-cadherin and beta-catenin in folate sufficient cells for both Caco-2 and FHC cells. Folate deficient Caco-2 cells showed no change in E-cadherin, but an increase in membrane associated beta-catenin. Folate deficient FHC cells showed an increase in nuclear localization of both E-cadherin and beta-catenin. The AKT pathway was also analyzed in Caco-2 cells to determine if alterations in this growth regulatory pathway was associated with folate deficiency. In Caco-2 cells, an increase in activated AKT was detected at day 30 in folate deficient cells, and other proteins in the AKT pathway (PDK1, PTEN, and c-Raf) showed slight decreases whereas a small increase was detected for GSK-3beta. Finally, we confirmed published findings that Caco-2 cells were adapting to folate deficiency by upregulating the folate transporters: folate receptor 1 (FolR-1) and reduced folate carrier (RFC). In general, those results suggest that alterations in E-cadherin and beta-catenin occur in folate deficient colon epithelial cells, although the cells can adapt and continue to proliferate.
机译：叶酸缺乏与多种健康问题有关，包括神经管缺陷和癌症，尤其是大肠癌。叶酸是DNA合成和细胞甲基化反应所需的基本饮食维生素。这些关联之间的机械联系尚不完全清楚。叶酸缺乏症已显示会影响与Wnt途径相关的基因，包括β-连环蛋白。 E-cadherin通过控制游离β-catenin的水平是Wnt信号的重要调节剂。还显示出E-钙粘蛋白受叶酸缺乏影响。这项研究的目的是确定叶酸缺乏症是否改变了两种结肠上皮细胞系中E-钙粘蛋白和RNA，β-catenin蛋白以及E-钙粘蛋白和β-catenin的细胞水平。 Caco-2和FHC细胞在叶酸充足且缺乏的培养基中生长15天和30天。在叶酸缺乏的细胞中，两种细胞系的细胞生长速率都降低了，尽管细胞继续生长。细胞周期阶段分布显示，Caco-2细胞的S期和G2 / M期增加，而FHC细胞则没有变化。叶酸缺乏的Caco-2细胞在第15和30天时E-钙粘蛋白和β-catenin蛋白水平略有下降。这与第30天时E-钙粘蛋白转录本的下降有关。FHC细胞在第15天显示出相似的结果，第30天时E-钙粘着蛋白的含量略有增加，β-catenin则没有变化。 Caco-2细胞的免疫荧光染色显示，在足以容纳Caco-2和FHC细胞的叶酸充足的细胞中，E-钙粘着蛋白和β-连环蛋白的细胞质弥散性染色。叶酸缺乏的Caco-2细胞在E-钙粘着蛋白上没有变化，但是在膜相关的β-连环蛋白上却有增加。叶酸缺乏的FHC细胞显示E-钙粘蛋白和β-连环蛋白的核定位增加。还对Caco-2细胞中的AKT途径进行了分析，以确定该生长调节途径的改变是否与叶酸缺乏有关。在Caco-2细胞中，在叶酸缺乏的细胞中，在第30天检测到活化的AKT增加，并且AKT途径中的其他蛋白质（PDK1，PTEN和c-Raf）显示略有降低，而在GSK- 3beta。最后，我们证实了已发表的发现，即Caco-2细胞通过上调叶酸转运蛋白：叶酸受体1（FolR-1）和减少的叶酸载体（RFC）来适应叶酸缺乏。通常，这些结果表明，叶酸缺乏的结肠上皮细胞中E-钙粘蛋白和β-连环蛋白发生了改变，尽管这些细胞可以适应并继续增殖。

著录项

  • 作者

    Cockman, Matthew.;

  • 作者单位

    The University of North Carolina at Greensboro.;

  • 授予单位 The University of North Carolina at Greensboro.;
  • 学科 Biology Cell.
  • 学位 M.S.
  • 年度 2009
  • 页码 64 p.
  • 总页数 64
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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