2,8-二羟基腺嘌呤晶体诱导培养的肾上皮细胞α -连环蛋白、整合素α 3、整合素β 6和PDGF-B。

Li Wang, Nandita Raikwar, Min Yang, Li Deng, James A McAteer, Peter J Stambrook, Amrik Sahota, Jay A Tischfield
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引用次数: 8

摘要

背景:纯合腺嘌呤磷酸核糖基转移酶(APRT)缺乏与2,8-二羟基腺嘌呤(DHA)肾结石有关。利用Aprt基因敲除小鼠的全肾RNA,我们之前发现DHA在肾脏的沉积会导致与组织损伤相关的基因表达的变化。为了确定这些变化的细胞基础,我们研究了暴露于DHA或草酸钙一水合物(COM)晶体的培养人肾(NHK-C)和非洲绿猴(BSC-1)上皮细胞的基因表达。方法:从处理过的细胞和未处理过的细胞中分离mRNA合成第一链cDNA,将其杂交到包含588个与各种生理和病理过程相关的基因的膜结合cDNA阵列。通过反转录PCR证实了基因表达的变化。结果:27%的阵列cdna在未经处理的NHK-C细胞中以不同水平相对于管家基因表达。NHK-C细胞暴露于DHA后,三种粘附分子(α -catenin、整合素α 3和整合素β 6)和血小板衍生生长因子B (PDGF-B)的表达升高。BSC-1细胞中粘附分子的表达也有所增加,但PDGF-B未见表达。COM晶体也刺激了这四个基因在NHK-C细胞中的表达,但与DHA相比,表达谱在数量上有所不同。结论:这些发现提示DHA晶体刺激肾上皮细胞特异性基因的表达,DHA诱导细胞损伤的途径可能与COM晶体相似。粘附分子和PDGF-B的诱导可能影响细胞-细胞或细胞-基质相互作用和/或改变肌动蛋白细胞骨架。这些改变可能最终导致晶体引起的肾损伤。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Induction of alpha-catenin, integrin alpha3, integrin beta6, and PDGF-B by 2,8-dihydroxyadenine crystals in cultured kidney epithelial cells.

Background: Homozygous adenine phosphoribosyltransferase (APRT) deficiency is associated with 2,8-dihydroxyadenine (DHA) nephrolithiasis. Using whole kidney RNA from Aprt knockout mice, we previously showed that the renal deposition of DHA leads to changes in the expression of genes involved in tissue injury. To determine the cellular basis for these changes, we investigated gene expression in cultured human kidney (NHK-C) and African green monkey (BSC-1) epithelial cells exposed to DHA or calcium oxalate monohydrate (COM) crystals.

Methods: First-strand cDNAs, synthesized from mRNA isolated from treated and untreated cells, were hybridized to membrane-bound cDNA arrays containing 588 genes associated with various physiological and pathological processes. Changes in gene expression were confirmed by reverse transcription PCR.

Results: Twenty-seven percent of the array cDNAs were expressed in untreated NHK-C cells at varying levels relative to a housekeeping gene. The expression of three adhesion molecules (alpha-catenin, integrin alpha3, and integrin beta6) and platelet-derived growth factor B (PDGF-B) was elevated following exposure of NHK-C cells to DHA. Increased expression of the adhesion molecules was also observed in BSC-1 cells, but PDGF-B expression could not be detected. COM crystals also stimulated the expression of these four genes in NHK-C cells, but the expression profile was quantitatively different compared with DHA.

Conclusions: These findings suggest that DHA crystals stimulate the expression of specific genes in kidney epithelial cells and that the pathways for DHA-induced cell injury may be similar to those for COM crystals. The induction of adhesion molecules and PDGF-B may affect cell-cell or cell-matrix interactions and/or alter the actin cytoskeleton. These alterations may ultimately contribute to crystal-induced renal injury.

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