Project description:Genome wide DNA methylation profiling of normal and squamous carcinoma of the cervix. The Illumina Infinium 27k Human DNA methylation Beadchip v1.2 was used to obtain DNA methylation profiles across approximately 27,000 CpGs in frozen cervical samples. Samples included 4 normal ectocervices (Ecto 1 to 4) and 5 squamous cell carcinoma of the cervix (SCC 1 to 5). In order to identify epigenetic biomarkers for early cervix cancer diagnosis, we performed a methylation screening using Illumina Infinium 27k Human DNA methylation Beadchip v1.2 of stem cell marker promoters during cervical carcinogenesis and demonstrated a strong hypermethylation of Undifferentiated cell Transcription Factor 1 (UTF1) promoter in squamous cell carcinoma (SCC) in comparison with normal ectocervix. Direct bisulfite pyrosequencing of DNA isolated from liquid-based cytological samples showed that UTF1 promoter methylation increases with lesion severity and is associated with higher expression. By RT-PCR, Western Blot and immunofluorescence, we demonstrated that UTF1 mRNA and protein are expressed in epithelial cancer cell lines, even in the absence of its two main described regulators Oct4A and Sox2. Methyl-Specific PCR experiments revealed that the inhibition of DNA methyltransferase by 5-aza-2’-deoxycytidine is associated with decreased UTF1 gene methylation and expression. These findings provide evidence that UTF1 promoter methylation profile might be a useful biomarker for cervix cancer diagnosis and raise the questions of its role during epithelial carcinogenesis and of the mechanisms involved in the regulation of its expression. Bisulfite converted DNA from the 9 samples were hybridised to the Illumina Infinium 27k Human Methylation Beadchip v1.2
Project description:Classically, there are two types of endometrial cancer, endometrioid adenocarcinoma (EAC), or Type I; and uterine papillary serous carcinoma (UPSC), or Type II. These two types of cancers exhibit distinct DNA methylation levels in promoters of many genes. In EAC, many tumor suppressor genes were silenced due to DNA hypermethylation at their promoter region. However, promoters of many of these genes remained unmethylated in UPSC. Here, we described complete DNA methylome maps of endometrioid adenocarcinoma, uterine papillary serous carcinoma, and normal endometrium, by applying a combined strategy of methylated DNA immunoprecipitation sequencing (MeDIP-seq) and methylation-sensitive restriction enzyme sequencing (MRE-seq). We took a complementary and orthogonal approach to identify DNA methylation changes unique to the two endometrial cancer subtypes in an unbiased fashion. We generated complete DNA methylome maps for endometrioid adenocarcinoma (EAC, three samples), uterine papillary serous carcinomas (UPSC, three samples), and normal endometrium (pooled samples) by integrating data from methylated DNA immunoprecipitation sequencing (MeDIP-seq) and methylation-sensitive restriction enzyme sequencing (MRE-seq).
Project description:Classically, there are two types of endometrial cancer, endometrioid adenocarcinoma (EAC), or Type I; and uterine papillary serous carcinoma (UPSC), or Type II. These two types of cancers exhibit distinct DNA methylation levels in promoters of many genes. In EAC, many tumor suppressor genes were silenced due to DNA hypermethylation at their promoter region. However, promoters of many of these genes remained unmethylated in UPSC. Here, we described complete DNA methylome maps of endometrioid adenocarcinoma, uterine papillary serous carcinoma, and normal endometrium, by applying a combined strategy of methylated DNA immunoprecipitation sequencing (MeDIP-seq) and methylation-sensitive restriction enzyme sequencing (MRE-seq).
Project description:131 patient-derived xenograft models were generated for non-small cell lung carcinoma and were profiled at the genome, transcriptome and proteome level by analysis of gene copy number variation, whole exome sequencing, DNA methylation, transcriptome, proteome and phospho(Tyr)-proteome. At the proteome level, the human tumor and murine stroma were discernible. Tumor proteome profiling resolved the known major histological subtypes and revealed 3 proteome subtypes (proteotypes) among adenocarcinoma and 2 in squamous cell carcinoma that were associated with distinct protein-phosphotyrosine signatures and patient survival. Stromal proteomes were similar between histological subtypes, but two adenocarcinoma proteotypes had distinct stromal proteomes. Proteotypes comprise tumor and stromal signatures of targetable biological pathways suggesting that patient stratification by proteome profiling may be an actionable approach to precisely diagnose and treat cancer.
Project description:The Illumina Infinium 27k Human DNA methylation Beadchip v1.2 was used to obtain DNA methylation profiles across approximately 27,000 CpGs in normal and cancer tissue from the uterine cervix. There are a total of 63 samples, 15 samples of normal histology and 48 cervical cancers. The normal cervical samples were from women who underwent a hysterectomy for uterine fibroids. The 48 cervical cancers were from women who were treated at the Innsbruck Medical University between 1990 and 2006.
Project description:Cancer of the uterine cervix (CACX) is the third most commonly diagnosed cancer and the fourth leading cause of cancer deaths in women worldwide. In India, CACX accounts for approximately 1,22,844 new cases and 67,477 deaths annually. Previously, we catalogued global copy-number aberrations [GSE76911] and performed gene expression profiling [GSE122697] in CACX. Interestingly, differential change in the expression between normal and tumor tissues of several genes did not correlate with the chromosomal copy-number alteration. This encouraged us to perform genome-wide DNA methylation analysis. Hence, in the current study, we discover the global methylation in cervical tumors at different clinical stages and HPV-negative normal ectocervix along with HPV16-positive cervical squamous cell carcinoma cell line, SiHa.
Project description:Utilizing methylation DNA immunoprecipitation (MeDIP) coupled with promoter tiling arrays, we analyzed the methylation profiles of pooled DNA from human cervical carcinoma and normal cervix Cervical carcinoma compaire with normal cervix
Project description:Lung cancer is the leading cause of preventable death globally and is broadly classified into adenocarcinoma and squamous cell carcinoma depending upon cell type. In this study, we carried out mass spectrometry based quantitative proteomic analysis of lung adenocarcinoma and squamous cell carcinoma primary tissue by employing the isobaric tags for relative and absolute quantitation (iTRAQ) approach. Proteomic data was analyzed using SEQUEST search algorithm which resulted in identification of 25,998 peptides corresponding to 4,342 proteins of which 610 proteins were differentially expressed (≥ 2-fold) between adenocarcinoma and squamous cell carcinoma samples. These differentially expressed proteins were further classified by gene ontology for their localizations and biological processes. Pathway analysis of differentially expressed proteins revealed distinct alterations in networks and pathways in both adenocarcinoma and squamous cell carcinoma samples. In this study, we identified a subset of proteins that shows converse expression between lung adenocarcinoma and squamous cell carcinoma samples. Such proteins may serve as signature markers to distinguish between the two subtypes.
Project description:Utilizing methylation DNA immunoprecipitation (MeDIP) coupled with promoter tiling arrays, we analyzed the methylation profiles of pooled DNA from human cervical carcinoma and normal cervix
Project description:The Illumina Infinium 27k Human DNA methylation Beadchip v1.2 was used to obtain DNA methylation profiles across approximately 27,000 CpGs in normal and cancer tissue from the uterine cervix. There are a total of 63 samples, 15 samples of normal histology and 48 cervical cancers. The normal cervical samples were from women who underwent a hysterectomy for uterine fibroids. The 48 cervical cancers were from women who were treated at the Innsbruck Medical University between 1990 and 2006. Bisulphite converted DNA from the 63 samples were hybridised to the Illumina Infinium 27k Human Methylation Beadchip v1.2