Project description:This laboratory focuses on selectin mediated recruitment during adoptive immunotherapy for metastatic cancer. This study seeks to determine changes in the expression levels of Fucosyltransferases, Selectins, and cytokines in normal and inflamed mouse skin, melanoma tumor tissue of different sizes, and tumor cells grown in culture. Since the ability to treat the tumor effectively is directly related to the size of the tumor, differences in glyco-expression patterns may be of interest. In this study, five groups were hybridized and analyzed using the GLYCOv2 array. Each group was analyzed in triplicate. The groups were: Normal mouse skin, normal mouse skin inflamed by treatment with Oxazolone, B16-OVA melanoma tissue from 6 day tumors, B16-OVA melanoma tissue from 11 day tumors, and B16-OVA grown in cell culture.
Project description:We previously showed that transgenic enhancement of histamine production in B16-F10 melanomas strongly supports tumor growth in C57Bl/6 mice (Cancer Res, 2005 Pos et al.). In this array experiment, gene expression profiles of transgenic mouse melanomas, secreting different amounts of histamine, were compared by whole genome microarrays. Experiment Overall Design: By modifying the levels of L-histidine decarboxylase (HDC), the sole enzyme responsible for histamine production, we introduced three novel variants of the B16-F10 mouse melanoma cell line, displaying diminished (B16-F10 HDC-A), unmodified (B16-F10 HDC-M) or enhanced (B16-F10 HDC-S) capacities to produce and secrete histamine. Experiment Overall Design: In this experiment, B16-F10 HDC-A, HDC-M, and HDC-S experimental mouse melanomas were compared by analyzing 6-6 tumors in each group. Experiment Overall Design: In order to reduce the amount of arrays required, equal amounts of randomly chosen RNA sample pairs were pooled in each group, thus, at the end, each group consisted of 3 pooled tumor samples. All samples were biological replicates, no technical replicates or dye swapping were done. Experiment Overall Design: Gene expression patterns of the three tumor groups were compared indirectly, via a common reference samplei n a two-color array design. Arrays shown here represent gene expression patterns of individual tumor samples compared to the reference sample.
Project description:To establish cancer cells escaping from host immunity, we used murine B16 melanoma cell line expressing ovalbumin (B16OVA) and inoculated into mice immunized with OVA. Using this model, we established cell lines after the in vivo passage through distinct immunological condition. B16OVA tumor exposed to OVA-specific CD8+ T cell immunity in OVA-immunized B6 mice were isolated and established five variants (IMM1, 2, 8). For comparison, B16OVA tumors from non-immunized naïve B6 mice or OVA-immunized IFN-gamma-deficient mice were also isolated and established four variants, namely NIMM (1, 3, 4) cell lines or GKO-IMM (1, 2, 3) cell lines, respectively.
Project description:PURPOSE: To provide a detailed gene expression profile of the normal postnatal mouse cornea. METHODS: Serial analysis of gene expression (SAGE) was performed on postnatal day (PN)9 and adult mouse (6 week) total corneas. The expression of selected genes was analyzed by in situ hybridization. RESULTS: A total of 64,272 PN9 and 62,206 adult tags were sequenced. Mouse corneal transcriptomes are composed of at least 19,544 and 18,509 unique mRNAs, respectively. One third of the unique tags were expressed at both stages, whereas a third was identified exclusively in PN9 or adult corneas. Three hundred thirty-four PN9 and 339 adult tags were enriched more than fivefold over other published nonocular libraries. Abundant transcripts were associated with metabolic functions, redox activities, and barrier integrity. Three members of the Ly-6/uPAR family whose functions are unknown in the cornea constitute more than 1% of the total mRNA. Aquaporin 5, epithelial membrane protein and glutathione-S-transferase (GST) omega-1, and GST alpha-4 mRNAs were preferentially expressed in distinct corneal epithelial layers, providing new markers for stratification. More than 200 tags were differentially expressed, of which 25 mediate transcription. CONCLUSIONS: In addition to providing a detailed profile of expressed genes in the PN9 and mature mouse cornea, the present SAGE data demonstrate dynamic changes in gene expression after eye opening and provide new probes for exploring corneal epithelial cell stratification, development, and function and for exploring the intricate relationship between programmed and environmentally induced gene expression in the cornea. Keywords: other
Project description:Analysis of gene expression patterns in cancer improved the understanding the mechanisms underlying the process of metastatic progression. Recent studies have attributed an important role to B-1 cells, a subset of B lymphocytes, in melanoma progression. It was already demonstrated that in vitro interaction between B16 melanoma cells and B-1 lymphocytes induced increase in metastatic potential of B16 lineage. In this study we used a microarray approach to assess gene expression profile in B16 melanoma cells after contacting B-1 lymphocytes (B16B1).