Comparative genomic hybridization by array of mouse Brca1 and Brca2 mammary tumors
Ontology highlight
ABSTRACT: Comparison of mammary tumors from 35 Brca1F/F;p53F/F, 62 Brca2F/F;p53F/F and 33 p53F/F mice. For each mouse, tumor DNA was hybridized with its spleen DNA as a reference.
Project description:Comparison of mammary tumors from 35 Brca1F/F;p53F/F, 62 Brca2F/F;p53F/F and 33 p53F/F mice. For each mouse, tumor DNA was hybridized with its spleen DNA as a reference.
Project description:Clonal cell lines derived from primary murine Small Cell Lung Cancer tumors where analyzed by Comperative genomic hybridisation to identify common regions.
Project description:WT or CD27-/- OT-I T cells are stimulated in vitro with artficial antigen presenting cell (aAPC) lines expressing CD70 and/or CD80 for 2, 4, 8 or 14 hours.
Project description:WT and CD137L-/- mice of 8-12 weeks old were immunized intra-peritoneally with 50 µg chicken gamma- globulin conjugated to 4-hydroxy-3-nitrophenylacetyl (NP-CG) in alum or infected intranasally with 25 hemagglutinin units of influenza virus strain A/NT/60/68 in 50 µl HBSS. Nine days after immunization or infection, B cells were enriched from pooled spleens and lymph nodes of 4 mice per test group by means of magnetic labelled bead cell separation (MACS) using anti-mouse BD ImagTM CD45R/B220-particles DM. The B220+ MACS-sorted populations were stained with anti-GL7-FITC and anti-CD19-PE to sort CD19+GL7+ GC and CD19+GL7- non-GC B lymphocytes by flow cytometry on a FACSAria (BD). Around 1 million cells of each sample were obtained. These cells were spun down and the pellet was frozen in liquid nitrogen and kept at -80 ºC before RNA extraction.
Project description:It is generally accepted that human cancers derive from a mutated single cell. However, the genetic steps characterizing various stages of progression remain unclear. Studying a unique case of metastatic melanoma, we observed that cell lines derived from metachronous metastases arising over a decade retained a central core of genetic stability in spite of divergent phenotypes. In the present study we expanded our previous observations comparing these autologous cell lines of clonal derivation with heterologous ones and correlated array Comparative Genomic Hybridization (aCGH) with gene expression profiling to determine their relative contribution to the dynamics of disease progression. aCGH and gene expression profiling were performed on autologous cell lines and heterologous melanoma cell lines originated from other patients. A striking correlation existed between total extent of genetic imbalances, global transcriptional patterns and cellular phenotypes; they did not follow a strict temporal progression but stemmed independently at various time points from a central core of genetic stability best explained according to the cancer stem cell hypothesis; although their contribution was intertwined, genomic imbalances detectable by aCGH contributed only 25% of the transcriptional traits determining autologous tumors distinctiveness. Our study provides important insights about the dynamics of cancer progression and supports the development of targeted anti-cancer therapies against stable genetic factors determining the individuality of each patientâs disease that are maintained throughout the end stage of disease. Keywords: genetic modification design PBMC from a female donor were Ficoll gradient separated and used throughout the study as reference.. Validation of array CGH accuracy was done by obtaining six additional PBMC from female donors and six PBMC from male donors to confirm stability of gene representation in autosomes and sex-determined imbalances within the X and Y chromosome regions. Five melanoma cell lines were generated from distinct cutaneous melanoma metastases that progressively appeared in patient 888. Other melanoma cell lines were generated from cutaneous melanoma metastases from other patients. The melanoma cell line A375 was purchased from the American Type Culture Collection. For each cell line arrayCGh has been performed.
Project description:The molecular chaperone Hsp90 is an important regulator of proteostasis. It has remained unclear why S. cerevisiae possesses two Hsp90 isoforms, the constitutively expressed Hsc82 and the stress-inducible Hsp82. Here, we report distinct differences despite a sequence identity of 97 %. Consistent with its function under stress conditions, Hsp82 is more stable and refolds more efficiently than Hsc82. Hspc82 and Hsc82 also differ in their ATPases and conformational cycles. Hsc82 is more processive and populates closed states to a greater extent. Variations in the N-terminal ATP-binding domain modulate its dynamics and conformational cycle. Despite these differences, the client interactomes, are largely identical, but isoform-specific interactors exist both under physiological conditions and heat shock. Taken together, changes mainly in the N-domain create a stress-specific, more resilient protein with a shifted activity profile. Thus, the precise tuning of the Hsp90 isoforms preserves the basic mechanism but adapts it to specific needs
Project description:Gene expression array analysis on a series of ten different histological special types of invasive breast carcinomas (tubular, micropapillary, mucinous A, mucinous B, endocrine, apocrine, metaplastic, medullary, adenoid cystic, invasive ductal carcinoma with osteoclastic giant cells) and invasive lobular carcinoma.<br><br>Note: this experiment was reloaded into ArrayExpress in August 2010 to include mappings between raw and processed data files. It now includes additional dye-swap combined normalized data files.