Subramanian Veedamali S, Reidling Jack C, Said Hamid M
Departments of Medicine, University of California, Irvine, California, USA.
Am J Physiol Cell Physiol. 2008 Sep;295(3):C828-35. doi: 10.1152/ajpcell.00249.2008. Epub 2008 Jul 23.
Differentiation of intestinal epithelial cells is accompanied by alterations in levels of expression of many genes, including those involved in nutrient uptake. Effects of differentiation of intestinal epithelial cells on the physiological and molecular parameters of the intestinal folate uptake process are not well characterized. To address this issue, we used two models, Caco-2 cells and native mouse intestine. Studies with Caco-2 cells showed a significant increase in the initial rate of carrier-mediated folic acid uptake during differentiation (i.e., as the cells transitioned from preconfluent to confluent and then to postconfluent stages). This increase was associated with an increase in the level of expression of the human reduced folate carrier (hRFC) and the human proton-coupled folate transporter (hPCFT) both at the protein and mRNA levels with differentiation; it was also associated with a significant increase in activity of the hRFC and hPCFT promoters. Studies with native mouse intestine showed a significantly higher folate uptake in villus compared with crypt cells, which was again associated with a significantly higher level of expression of the mouse RFC and PCFT at the protein and mRNA levels. Together, these studies demonstrate that the intestinal folate uptake process undergoes differentiation-dependent regulation and that this regulation is mediated via changes in the level of expression of both the RFC and PCFT. In addition, the studies suggest the possible involvement (at least in part) of a transcriptional mechanism(s) in this type of regulation of the intestinal folate uptake process.
肠道上皮细胞的分化伴随着许多基因表达水平的改变,包括那些参与营养物质摄取的基因。肠道上皮细胞分化对肠道叶酸摄取过程的生理和分子参数的影响尚未得到充分表征。为了解决这个问题,我们使用了两种模型,即Caco-2细胞和天然小鼠肠道。对Caco-2细胞的研究表明,在分化过程中(即当细胞从汇合前过渡到汇合然后到汇合后阶段时),载体介导的叶酸摄取的初始速率显著增加。这种增加与人类还原型叶酸载体(hRFC)和人类质子偶联叶酸转运体(hPCFT)在蛋白质和mRNA水平上的表达水平随分化而增加有关;它还与hRFC和hPCFT启动子的活性显著增加有关。对天然小鼠肠道的研究表明,与隐窝细胞相比,绒毛中的叶酸摄取显著更高,这同样与小鼠RFC和PCFT在蛋白质和mRNA水平上的显著更高的表达水平有关。总之,这些研究表明肠道叶酸摄取过程经历了分化依赖性调节,并且这种调节是通过RFC和PCFT表达水平的变化介导的。此外,这些研究表明转录机制(至少部分)可能参与了肠道叶酸摄取过程的这种调节类型。