Abstract 3995: Quantitative protein profiling of tumor angiogenesis and metastasis biomarkers in mouse and human models

Abstract

Abstract Tumor and stromal cells secrete a variety of proteins acting as extracellular signals and creating a supportive microenvironment for tumor development, angiogenesis, and metastasis. We used the MILLIPLEX MAP cytokine/chemokine, cancer, angiogenesis, metastasis, MMP, bone metabolism, adipocyte, and cell signaling multi-pathway panels, based on the Luminex xMAP technology, to quantitatively evaluate the protein biomarker expression profiles on cultured mouse and human tumor cells and stromal cells as well as in vivo mouse and human tumor tissues. In the mouse system, biomarker profiling was performed to characterize secreted protein levels in Lewis Lung carcinoma (LLC) cells, B16.10 melanoma cells, b.End3 endothelial cells, and LLC tumors. The in vivo characterization, using primary tumors from mice bearing metastatic LLC, has identified differential expression of a panel of biomarkers, including amphiregulin (AREG), EGF, PECAM-1, KC, MCP-1, MIP-2, MMP-12, and resistin, between the high-fat (45% fat) and AIN-93G (15% fat) fed mice (p<0.05, 8 mice per group). These putative biomarkers may be involved in molecular mechanisms of the high-fat enhanced metastasis. Cell signaling pathway analysis revealed the activation of multiple pathways (JNK, p38, ERK, and STAT) in this mouse LLC model. In the human system, a heat map was generated to summarize the secreted protein signatures and the soluble receptor tyrosine kinases (RTK) profile on a series of the cultured human tumor cells (lung, brain, breast, prostate, liver, and colon) and 3 other human cell lines, HUVEC (endothelial), THP-1 (monocyte), and HFF-1 (fibroblast). Lastly, expression profiling of tumor and adjacent normal tissues from 2 patients with metastatic breast cancer was analyzed. Elevated levels of protein biomarkers, including MDC, IL-1RA, IL-8, IP-10, MCP-1, MIP-1α, OPG, OPN, SPARC, periostin, endoglin, PLGF, VEGF-A, MMP-1, MMP-7, MMP-9, and MMP-12, were detected in the breast tumors analyzed compared to their matched adjacent normal breast tissue. In summary, our results demonstrated that multiplexing using the Luminex xMAP bead-based platform for simultaneous quantitative expression profiling is a useful proteomic research tool, providing insights into biomarkers related to tumor progression and metastasis. Citation Format: Wen-Rong Lie, Jonathan Lipsey, Tim Warmke, Lin Yan, Jehangir Mistry. Quantitative protein profiling of tumor angiogenesis and metastasis biomarkers in mouse and human models. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3995. doi:10.1158/1538-7445.AM2014-3995