Project description:3 months old untreated male/female animals were used in the experiment. We used microarray analysis to determine differential gene expression in mice livers overexpressing WT K8 and K8 variants.
Project description:3 months old untreated male/female animals were used in the experiment. We used microarray analysis to determine differential gene expression in mice livers overexpressing WT K8 and K8 variants. Mice liver tissues were used for RNA extraction and hybridization on Affymetrix microarrays.
Project description:The epithelial specific keratin pair, Keratin 8/18 (K8/18), has been reported to be aberrantly expressed in squamous cell carcinoma (SCC) which is also correlated with increased invasiveness and poor prognosis. A Majority of Keratin 8 (K8) functions are governed by its phosphorylation at Serine73 (head domain) and Serine431 (tail domain) residues. Although deregulation of K8 phosphorylation has been associated with progression of different carcinomas, its role in skin SCC and the underlying mechanism is obscured. To understand the molecular basis of K8 phosphorylation mediated regulation of skin SCC progression, we performed TMT-based quantitative phosphoproteomics by expressing K8 wild type, phosphodead and phosphomimetic mutants in K8 deficient A431 cells. Bioinformatic analysis of our phosphoproteomic data showed differential regulation of phosphoproteins associated with migratory, proliferative and invasive potential in these cells. Further validation of protein phosphorylation levels and phenotypic validation of our phosphoproteomic data suggested potential role of K8 site specific phosphorylation/dephosphorylation in neoplastic progression of A431 cells.
Project description:BRCA1 mutation-carriers are predisposed to develop Basal-like breast cancer (BLBC), and p53 mutations are present in the majority of human BLBC cases, suggesting loss of these two tumor suppressors play key roles in development of BLBC. Recent studies suggest that the majority of human breast cancers, including BLBC, may originate from mammary epithelial cells (MECs) in the luminal lineage. However, how loss of p53 and BRCA1 contributes to development of BLBC from luminal MECs remains largely elusive. We developed a novel genetic targeting and lineage tracing approach based on intraductal injection of Cre-expressing adenovirus under the control of the pan-luminal Keratin 8 (K8) promoter (Ad-K8-Cre). We performed intraductal injection of Ad-K8-Cre to female mice carrying conditional knockout alleles of Brca1 and Trp53. The injected females developed mammary tumors within 12 months after injection. Microarray expression profiling of these tumors showed that they most closely resembled human BLBC.
Project description:Dkk4 transgenic mice in wild-type background showed abnormal hair subtype formation. Gene expression profiling showed significant down regulation of hair follicle-related keratin genes and hair keratin-associated protein genes at E18.5 and P1. Keywords: development or differentiation design,genetic modification design
Project description:BRCA1 mutation-carriers are predisposed to develop Basal-like breast cancer (BLBC), and p53 mutations are present in the majority of human BLBC cases, suggesting loss of these two tumor suppressors play key roles in development of BLBC. Recent studies suggest that the majority of human breast cancers, including BLBC, may originate from mammary epithelial cells (MECs) in the luminal lineage. However, how loss of p53 and BRCA1 contributes to development of BLBC from luminal MECs remains largely elusive. We developed a novel genetic targeting and lineage tracing approach based on intraductal injection of Cre-expressing adenovirus under the control of the pan-luminal Keratin 8 (K8) promoter (Ad-K8-Cre). We performed intraductal injection of Ad-K8-Cre to female mice carrying conditional knockout alleles of Brca1 (Brca1L) and Trp53 (Trp53L). The injected females developed mammary tumors similar to human BLBC within 12 months after injection. Here we characterized MECs targeted by Ad-K8-Cre one month after the intraductal injection.
Project description:BRCA1 mutation-carriers are predisposed to develop Basal-like breast cancer (BLBC), and p53 mutations are present in the majority of human BLBC cases, suggesting loss of these two tumor suppressors play key roles in development of BLBC. Recent studies suggest that the majority of human breast cancers, including BLBC, may originate from mammary epithelial cells (MECs) in the luminal lineage. However, how loss of p53 and BRCA1 contributes to development of BLBC from luminal MECs remains largely elusive. We developed a novel genetic targeting and lineage tracing approach based on intraductal injection of Cre-expressing adenovirus under the control of the pan-luminal Keratin 8 (K8) promoter (Ad-K8-Cre). We performed intraductal injection of Ad-K8-Cre to female mice carrying conditional knockout alleles of Brca1 (Brca1L) and Trp53 (Trp53L). The injected females developed mammary tumors similar to human BLBC within 12 months after injection. Here we characterized MECs targeted by Ad-K8-Cre at different time points after the intraductal injection, as well as mammary tumors developed in this model, by single cell expression analysis.
