Keay Susan, Seillier-Moiseiwitsch Francoise, Zhang Chen-Ou, Chai Toby C, Zhang Jialu
Division of Infectious Diseases, Department of Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Physiol Genomics. 2003 Jul 7;14(2):107-15. doi: 10.1152/physiolgenomics.00055.2003.
Explanted bladder epithelial cells from patients with interstitial cystitis (IC) have been shown to differ from explanted control cells in several ways, including production of an antiproliferative factor (APF), altered production of certain epithelial growth factors, and rate of proliferation. To better understand the role of the APF in abnormal bladder epithelial cell proliferation in IC, we studied gene expression patterns in normal bladder epithelial cells treated with APF vs. mock APF and compared them to expression patterns in IC vs. normal cells using microarray analysis. Oligo-dT-primed total cellular RNA was labeled with [(33)P]dCTP and hybridized to GeneFilter GF211 microarray membranes (Research Genetics) containing cDNA for 3,964 human genes. Thirteen genes that function in epithelial cell proliferation or differentiation were consistently differentially expressed in both IC (compared with control) and APF-treated (compared with mock APF-treated) normal bladder epithelial cells. The general pattern of gene expression in IC and APF-treated cells suggested a less proliferative phenotype, with increased expression of E-cadherin, phosphoribosylpyrophosphate synthetase-associated protein 39, and SWI/SNF complex 170-kDa subunit, and decreased expression of vimentin, alpha2-integrin, alpha1-catenin, cyclin D1, and jun N-terminal kinase 1; these findings were confirmed for the structural gene products (E-cadherin, vimentin, alpha2-integrin, and alpha-catenin) by immunohistochemistry. These results are compatible with the previously noted decreased proliferation rate of IC and APF-treated normal cells, and indicate that the mechanism whereby APF inhibits cell proliferation may involve both downregulation of genes that stimulate cell proliferation along with upregulation of genes that inhibit cell growth.
已证实，间质性膀胱炎（IC）患者的移植膀胱上皮细胞在多个方面与移植的对照细胞不同，包括抗增殖因子（APF）的产生、某些上皮生长因子产生的改变以及增殖速率。为了更好地理解APF在IC中膀胱上皮细胞异常增殖中的作用，我们研究了用APF处理的正常膀胱上皮细胞与模拟APF处理的正常膀胱上皮细胞的基因表达模式，并使用微阵列分析将它们与IC细胞和正常细胞的表达模式进行比较。用寡聚dT引物合成的总细胞RNA用[³³P]dCTP标记，并与包含3964个人类基因cDNA的GeneFilter GF211微阵列膜（Research Genetics）杂交。在IC（与对照相比）和APF处理的（与模拟APF处理相比）正常膀胱上皮细胞中，13个在上皮细胞增殖或分化中起作用的基因始终存在差异表达。IC细胞和APF处理细胞中的基因表达总体模式表明其增殖表型减弱，E-钙黏蛋白、磷酸核糖焦磷酸合成酶相关蛋白39和SWI/SNF复合物170-kDa亚基的表达增加，波形蛋白、α2整合素、α1连环蛋白、细胞周期蛋白D1和Jun N端激酶1的表达减少；免疫组织化学证实了这些结构基因产物（E-钙黏蛋白、波形蛋白、α2整合素和α连环蛋白）的这些发现。这些结果与之前观察到的IC细胞和APF处理的正常细胞增殖速率降低一致，并表明APF抑制细胞增殖的机制可能涉及刺激细胞增殖的基因的下调以及抑制细胞生长的基因的上调。
