The role of differential glyco-gene expression in cancer metastasis
Lum, Michelle A
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Metastasis involves invasion, intravasation, transport, arrest, extravasation, secondary growth, and angiogenesis. To study carbohydrate-mediated interactions in these processes, four closely-related murine mammary carcinoma cell lines were used: metastatic 4T1 and 4T1 altered-properties, and non-metastatic 4T07 and 67NR. The Consortium for Functional Glycomics (CFG) used RNA from each cell line to compare carbohydrate-related gene expression on a Glyco-gene chip. The Glyco-gene chip is a microarray capable of detecting over 1,800 different carbohydrate-related genes. Thirteen differentially-expressed genes on the Glyco-gene chip were validated by quantitative RT-PCR. Flow cytometry was used to detect cell surface expression of differentially-expressed gene products including: VCAM-1, CD34, TF-antigen, and linkages of sialic acid and carbohydrates. The CFG performed glycan profiling on each cell line. Cell surface carbohydrates were identified and quantified using a technique similar to mass-spectrometry. This analysis can be utilized to further understand, and to develop therapy for the prevention of metastasis.