Institute for Bioengineering and Nanoscience in Advanced Medicine, Northwestern University, Evanston, IL, USA.
Clin Exp Metastasis. 2009;26(8):993-1004. doi: 10.1007/s10585-009-9289-0. Epub 2009 Sep 29.
After prostate cancer cells (PCa) arrive in bone, interactions with cells that include long bone osteoblasts (LBOB) and bone marrow stromal cells (BMSC) lead to metastasis formation. The effect of heterotypic cell-cell contact between PCa cells and BMSC or LBOB on PCa cell gene expression is poorly understood. To establish the role of heterotypic contact in bone metastasis formation, we mixed and co-cultured PC3 cells with rat BMSC, LBOB, or human prostate stromal cells (PS15). PC3 cells were then re-isolated for gene array analysis, and imaged using in situ hybridization to confirm that heterotypic contact regulates gene expression. The gene expression was examined using focused gene arrays containing 96 each of tumor metastasis genes or osteogenesis genes. A total of 18 out of 192 genes in PC3 cells were found to be under or over expressed subsequent to heterotypic contact with BMSC when analyzed. A total of 15 genes out of 192 were regulated in co-culture with LBOB, and 19 genes with PS15. Only two genes, uPA and Collagen III, were regulated by contact with BMSC or LBOB (both are bone derived cells), but not by contact with PS15. The relationship between cell-cell contact and uPA expression was further explored by varying cell ratios in co-culture. uPA over-expression in PC3 was related to the BMSC:PC3 ratio, and was maximum at a 10:1 ratio, where most PC3 cells would be in contact with BMSC, as predicted by a theoretical model of heterotypic contact. In situ staining of micropatterned PC3 and BMSC cells showed that uPA over-expression localizes to regions of heterotypic cell-cell contact. Taken together, our results suggest that heterotypic cell-to-cell contact between PC3 and BMSC proportionally enhances gene expression for uPA, providing a mechanism for localized control of invasiveness.
前列腺癌细胞 (PCa) 转移到骨骼后,与包括长骨成骨细胞 (LBOB) 和骨髓基质细胞 (BMSC) 在内的细胞的异质细胞-细胞相互作用导致转移形成。PCa 细胞与 BMSC 或 LBOB 之间的异质细胞-细胞接触对 PCa 细胞基因表达的影响知之甚少。为了确定异质接触在骨转移形成中的作用,我们将 PC3 细胞与大鼠 BMSC、LBOB 或人前列腺基质细胞 (PS15) 混合并共培养。然后将 PC3 细胞重新分离进行基因芯片分析,并通过原位杂交进行成像以确认异质接触调节基因表达。使用包含 96 个肿瘤转移基因或成骨基因的聚焦基因芯片检查基因表达。在与 BMSC 异质接触后,192 个 PC3 细胞中的 18 个基因被发现表达下调或上调。在与 LBOB 共培养中,192 个基因中有 15 个受到调节,与 PS15 共培养中有 19 个基因受到调节。只有两个基因,uPA 和 Collagen III,受到 BMSC 或 LBOB(均为骨骼来源细胞)的接触调节,但不受 PS15 的调节。通过改变共培养中的细胞比例进一步探索细胞-细胞接触与 uPA 表达之间的关系。PC3 中的 uPA 过表达与 BMSC:PC3 比例有关,在 10:1 比例时最大,在这种比例下,大多数 PC3 细胞将与 BMSC 接触,这与异质接触的理论模型预测一致。对微图案化的 PC3 和 BMSC 细胞进行原位染色显示,uPA 过表达定位于异质细胞-细胞接触的区域。综上所述,我们的结果表明,PC3 和 BMSC 之间的异质细胞-细胞接触成比例地增强了 uPA 的基因表达,为局部控制侵袭性提供了一种机制。