Calaluce R, Kunkel M W, Watts G S, Schmelz M, Hao J, Barrera J, Gleason-Guzman M, Isett R, Fitchmun M, Bowden G T, Cress A E, Futscher B W, Nagle R B
Arizona Cancer Center, University of Arizona Health Sciences Center, Tucson, Arizona 85724-5043, USA.
Mol Carcinog. 2001 Feb;30(2):119-29. doi: 10.1002/1098-2744(200102)30:2<119::aid-mc1020>3.0.co;2-n.
Interactions between extracellular matrix (ECM) proteins and prostate carcinoma cells provide a dynamic model of prostate tumor progression. Previous work in our laboratory showed that laminin-5, an important member of a family of ECM glycoproteins expressed in the basal lamina, is lost in prostate carcinoma. Moreover, we showed that the receptor for laminin-5, the alpha6beta4 integrin, is altered in prostate tumors. However, the genes that laminin-5 potentially regulates and the significance of its loss of expression in prostate cancer are not known. We selected cDNA microarray as a comprehensive and systematic method for surveying and examining gene expression induced by laminin-5. To establish a definitive role for laminin-5 in prostate tumor progression and understand the significance of its loss of expression, we used a cDNA microarray containing 5289 human genes to detect perturbations of gene expression when DU145 prostate carcinoma cells interacted with purified laminin-5 after 0.5, 6, and 24 h. Triplicate experiments showed modulations of four, 61, and 14 genes at 0.5, 6, and 24 h, respectively. Genes associated with signal transduction, cell adhesion, the cell cycle, and cell structure were identified and validated by northern blot analysis. Protein expression was further assessed by immunohistochemistry. Mol. Carcinog. 30:119-129, 2001.
细胞外基质（ECM）蛋白与前列腺癌细胞之间的相互作用为前列腺肿瘤进展提供了一个动态模型。我们实验室之前的研究表明，层粘连蛋白-5是在基底层表达的ECM糖蛋白家族的重要成员，在前列腺癌中缺失。此外，我们还表明，层粘连蛋白-5的受体α6β4整合素在前列腺肿瘤中发生了改变。然而，层粘连蛋白-5可能调控的基因及其在前列腺癌中表达缺失的意义尚不清楚。我们选择cDNA微阵列作为一种全面系统的方法来检测和研究层粘连蛋白-5诱导的基因表达。为了确定层粘连蛋白-5在前列腺肿瘤进展中的明确作用并理解其表达缺失的意义，我们使用了一个包含5289个人类基因的cDNA微阵列，来检测DU145前列腺癌细胞在与纯化的层粘连蛋白-5相互作用0.5小时、6小时和24小时后基因表达的扰动情况。三次重复实验分别显示在0.5小时、6小时和24小时有4个、61个和14个基因发生了调节。通过Northern印迹分析鉴定并验证了与信号转导、细胞黏附、细胞周期和细胞结构相关的基因。通过免疫组织化学进一步评估了蛋白表达情况。《分子致癌学》30:119 - 129, 2001年。
