Project description:Prostate cancer (PCa) tends to be more aggressive and lethal in African Americans (AA) compared to European Americans (EA). To further understand the thebiological risk factors associated with PCa disparities observed in AA and EA patients, we performed microRNA profiling using Agilent Human miRNA arrays to identify the differentially expressed microRNAs beween: 1) AA and EA PCa patients; 2) AA PCa vs. AA normal; and 3) EA PCa vs. EA normal. 54 prostate biopsy specimens (tumor and adjacent normal tissues) were collected from 14 African American and 13 European American prostate cancer patients. 54 RNA samples, purified from the collected biopy specimens using Qiagen miRNeasy kit, were process and applied to Agilent human miRNA arrays. Array data was normalized and analyzed using Agilent GeneSpring program.
Project description:Prostate cancer (PCa) is a common cancer and remains the second leading cause of cancer-associated mortality in men. To investigate the involvement of differentially expressing genes in PCa with deregulated pathways to allow earlier diagnosis of the disease, transcriptomic analyses of differential expression genes in Fine-Needle Aspiration (FNA) biopsies were for discrimination of PCa from benign prostatic hyperplasia (BPH). The RNA samples were extracted from four PCa biopsy samples and four BPH biopsy controls for microarray profiling and performed in Affymetrix Human U133 Plus 2 arrays for gene expression profiling analysis. Microarray data were analyzed using GeneSpring GX 10 (Agilent).On average, we detected expression of 47,000 transcripts.Under the criteria fold change > 2 or < 0.5, we obtained 1819 differential expressed genes(DEGs).Hierarchy cluster analysis also indicated that the 8 samples were distributed into two clusters, 4 PCa samples in one cluster and 4 BPH samples in another cluster.Then,qRT-PCR validation of the DEGs in PCa tissue and prostate cancer cells.The results revealed that grouping was reasonable and the data can be directly applied to further analysis.
Project description:Comparison of miRNA expression profiles in normal and malignant prostate tissues. Keywords: microarray analysis of microRNA expression profiles MicroRNA expression was compared between normal prostate tissue from either young subjects that died of trauma, or normal adjacent to tumor, and prostatic tumors in older prostate cancer patients. RNA was isolated from frozen tissue sections, enriched for the miRNA fraction, which was subsequently labeled and hybridized to miRNA microarrays for expression profiling analysis.
Project description:Laser Capture Microdissected cells from archival FFPE Prostate cancer and normal adjacent tissues from 10 patients (5 CA and 5 AA) were converted to cDNA and analysed by PCR array to identify differentially expressed miRNAs between the groups. Selected differentially expressed miRNAs were further validated in tissues from 40 prostate cancer patients. The miRNAs which were differentially modulated in PCa patients in the validation set were further analysed in 32 urine samples from PCa patients and compared with 12 healthy individuals. Differentially expressed miRNAs were explored to e used as non-invasive biomarker for PCa. qPCR miRNA expression profiling. mRNA from 10 Prostate Cancer patients (5 Caucasian American and 5 African American) and their paired adjacent normal tissue were analysed to identify the differentially expressed miRNA between the groups. Equal amount small RNA from each group was pooled prior to gene expression analysis.
Project description:Prostate cancer (PCa) tends to be more aggressive and lethal in African Americans (AA) compared to European Americans (EA). To further understand the thebiological risk factors associated with PCa disparities observed in AA and EA patients, we performed microRNA profiling using Agilent Human miRNA arrays to identify the differentially expressed microRNAs beween: 1) AA and EA PCa patients; 2) AA PCa vs. AA normal; and 3) EA PCa vs. EA normal.
Project description:MicroRNA (miRNA) expression profiles were generated from prostate epithelial sub-populations, enriched from patient-derived benign prostatic hyperplasia (BPH, n=5), Gleason 7 treatment naïve prostate cancer (PCa, n=5) and castration-resistant prostate cancer (CRPC, n=3). Microarray expression was validated in an independent patient cohort (n=10). The analyses show that miRNA expression in epithelial sub-populations (e.g. stem cells) clustered together, irrespective of pathological status. We also discovered concordance between the miRNA expression profiles of unfractioned CRPCs, human embryonic stem cells (SCs), and prostate epithelial SCs (both, benign and malignant). miR-548c-3p was chosen as a candidate miRNA from this group to explore its utility as a CRPC biomarker and/or therapeutic target. Overexpression of miR-548c-3p was confirmed in cancer SCs (8-fold, p<0.05) and unfractionated CRPCs (1.8-fold, p<0.05). Enforced overexpression of miR-548c-3p in differentiated cells induced stem-like properties (p 0.01) and radioresistance (p<0.01). Re-analyses of published studies further revealed that miR-548c-3p is significantly overexpressed in CRPC (p<0.05) and is associated with poor recurrence-free survival (p<0.05), suggesting that miR-548c-3p is a functional biomarker for prostate cancer aggressiveness. Our results validate the prognostic and therapeutic relevance of miRNAs for advanced prostate cancer management, whilst demonstrating that resolving cell-type and differentiation-specific differences is essential to obtain clinical relevant miRNA expression profiles. There are 42 samples in total: 15 benign prostatic hyperplasia, 15 prostate cancers, 9 treatment-resistant prostate cancer. There are stem cells, transit amplifying cells and committed basal cells dissected from each sample. Three PrEC samples of each three cell types can be used as reference.