Repeated exposure to acid and bile selectively induces colonic phenotype expression in a heterogeneous Barrett's epithelial cell line
Repeated exposure to acid and bile selectively induces colonic phenotype expression in a heterogeneous Barrett's epithelial cell line
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DOI：
10.1038/labinvest.2008.34
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发表时间：
2008-06-01
影响因子：
5
通讯作者：
Das, Kiron M.
中科院分区：
文献类型：
--
作者：
Bajpai, Manisha;Liu, Jianying;Das, Kiron M.
Barrett上皮是食管远端的癌前特化柱状化生。我们证明了在非肿瘤性巴雷特细胞系（BAR-T）的细胞表型的变化，暴露于酸和胆汁盐，胃食管反流的两个重要组成部分。通过荧光激活细胞分选（FACS）使用针对以下标志物的单克隆抗体定量BAR-T细胞系中的细胞表型：细胞角蛋白8/18（CK 8/18）用于柱状细胞，CK 4用于鳞状细胞，mAbDas-1用于结肠上皮细胞表型，以及p75 NTR用于食管祖细胞。每天将细胞暴露于pH 4、pH 6和pH 7.4的200 mM甘氨鹅去氧胆酸或仅暴露于酸（pH 4）5分钟，持续长达6周。BAR-T细胞系包含35 +/-5.2%CK8/18、32 +/-3.5%mAbDas-1、9.5 +/-3%CK4和4 +/-2.5%p75NTR阳性细胞。单次暴露于酸和/或胆汁没有改变细胞表型。然而，至少2周的慢性治疗显著增强（P < 0.05）结肠表型和CK 8/18阳性细胞的表达，如通过FACS分析所证明的。pH 4的胆盐和胆盐+酸（pH 4）对结肠表型细胞的诱导作用最强（P < 0.01）。鳞状细胞（CK 4（+））表型没有改变的治疗。考克斯-2的表达诱导急性治疗后，增加到两倍，在慢性治疗过程中，特别是在响应酸性pH值。我们得出结论，BAR-T细胞可以作为一个“在体外”模型来研究环境因素的影响和它们对细胞表型和分子的食管癌发病机制的变化。
Barrett's epithelium is a precancerous, specialized columnar metaplasia in the distal esophagus. We demonstrate the changes in cellular phenotype in a non-neoplastic Barrett's cell line (BAR-T), following exposure to acid and bile salt, the two important components of gastroesophageal refluxate. Cell phenotypes in BAR-T cell line were quantified by fluorescence-activated cell sorting (FACS) using monoclonal antibodies against markers: cytokeratin 8/18 (CK8/18) for columnar, CK4 for squamous, mAbDas-1 for colonic epithelial cell phenotype and p75NTR for esophageal progenitors. Cells were exposed for 5 min each day to 200 mM glycochenodeoxycholic acid at pH 4, pH 6 and pH 7.4 or only to acid (pH 4) for up to 6 weeks. The BAR-T cell line comprised 35 +/- 5.2% CK8/18, 32 +/- 3.5% mAbDas-1, 9.5 +/- 3% CK4 and 4 +/- 2.5% p75NTR-positive cells. Single exposure to acid and or bile did not change cell phenotypes. However, chronic treatment for at least 2 weeks significantly enhanced (P < 0.05) the expression of colonic phenotype and CK8/18-positive cells, as evidenced by FACS analysis. Bile salt at pH 4 and bile salt followed by acid ( pH 4) in succession were the strongest stimulators (P < 0.01) for induction of colonic phenotype cells. Squamous (CK4(+)) phenotype did not change by the treatments. Cox-2 expression was induced after acute treatment and increased to twofold during chronic treatment, particularly in response to acidic pH. We conclude that BAR-T cells can be utilized as an 'in vitro' model to study the effect of environmental factors and their influence on the cellular phenotype and molecular changes in the pathogenesis of esophageal cancer.