MicroRNA profiling by array of human stage I epithelial ovarian cancer
ABSTRACT: In an effort to define the biological determinants of stage I chemo-sensitivity, we undertook a genome-wide miRNA expression profile in a cohort of 89 stage I patients (24 of which relapsed) with a complete follow up of twelve years, selected from a single tumor tissue collection of more than 1300 snap frozen tumor biopsies. With expression profile results for these samples and associated clinical variables, we investigated the association between miRNA and prognosis in stage I EOC. (Note: NED - No evidence of disease, DOD - Died of disease and DOC - Died of other causes)
Project description:In the last decades platinum-based neo-adjuvant chemotherapy (NACT) has been recognized as a reliable therapeutic strategy in patients with un-resectable advanced epithelial ovarian cancer (EOC). However, the molecular changes induced by NACT at miRNA level, and their prognostic role has not been clarified until now. In order to uncover miRNAs that are altered in EOC tumor which received NACT, we performed whole-miRNA analysis on 82 FIGO Stage IIIC-IV high-grade serous (HGS) tumors, whose samples had been collected at complete primary debulking (PDS) and at interval-debulking surgery (IDS) after fter 4 courses of NACT.
Project description:This study aims at correlating changes in the microRNA state in high grade serous epithelial ovarian cancer (HGS-EOC) to the response to therapy, in particular the insurgence of resistance to platinum-based treatment.
Project description:Downregulation of specific microRNAs contribute to epithelial-mesenchymal transition of Wilms' tumor cancer initiating cells. In order to gain insight into the biology of initiating cells/cancer stem cells (CIC/CSCs) in Wilms' tumor, we compared the microRNA expression profile of un-sorted propagatable WT xenografts (p-WT Xn), p-WT NCAM+ALDH1+-derived Xn and human fetal kidneys (hFKs). Global microRNA expression analysis showed specific microRNAs to differentially express identifying a strong miRNA signature for the NCAM+ALDH1+ WT CICs. 3 different tissue types (described in 'summary') each with n=4 were homoginized and analyzed for their global miRNA expression profiles. Human Fetal Kidney tissues were used as a control.
Project description:We measured the expression of human microRNAs in tumor cells derived from 8 FFPE samples among non-invasive CRC patients with different prognosis. Patients lived for more than 5 years after surgery were classified as good prognosis, and patients died within 5 years from surgery were classified as poor prognosis. Tumor tissues were macrodissected under the control HE staining slides. And there were at least 75 percent cancer cells in the samples. MicroRNA microarray was used to measure the expression of human miRNAs in tumor cells derived from 8 FFPE samples among early stage CRC patients with different prognosis.
Project description:A set of 45 surgical specimens has been profiled for miRNA expression to validate miRNA alterations associated to early relapse in advanced stage ovarian cancer patients. Fresh frozen samples were collected from a series of consecutive patients with high-grade advanced stage ovarian cancer who underwent primary surgery at INT-Milan. After surgery all patients received postoperative platinum-based chemotherapy. All patients signed an Institutional Review Board approved consent for bio-banking, clinical data collection and molecular analysis. Clinical codes: Histotype: according to WHO classification guidelines Stage: according to International Federation of Gynecological and Obstetrics (FIGO) guidelines Grading: according to WHO classification guidelines Debulking: NED: not evident disease; mRD: minimal residual disease; GRD: gross residual disease Therapy code: P: Platinum without taxanes; PT: Platinum/paclitaxel
Project description:Background: MicroRNA (miRNA) is an emerging subclass of small non-coding RNAs that regulates gene expression and has a pivotal role for many physiological processes including cancer development. Recent reports revealed the role of miRNAs as ideal biomarkers and therapeutic targets due to their tissue- or disease-specific nature. Head and neck cancer (HNC) is a major cause of cancer-related mortality and morbidity, and laryngeal cancer has the highest incidence in it. However, the molecular mechanisms involved in laryngeal cancer development remain to be known and highly sensitive biomarkers and novel promising therapy is necessary. Methodology/Principal Findings: To explore laryngeal cancer-specific miRNAs, RNA from 5 laryngeal surgical specimens including cancer and non-cancer tissues were hybridized to microarray carrying 723 human miRNAs. The resultant differentially expressed miRNAs were further tested by using quantitative real time PCR (qRT-PCR) on 43 laryngeal tissue samples including cancers, noncancerous counterparts, benign diseases and precancerous dysplasias. Significant expressional differences between matched pairs were reproduced in miR-133b, miR-455-5p, and miR-196a, among which miR-196a being the most promising cancer biomarker as validated by qRT-PCR analyses on additional 84 tissue samples. Deep sequencing analysis revealed both quantitative and qualitative deviation of miR-196a isomiR expression in laryngeal cancer. In situ hybridization confirmed laryngeal cancer-specific expression of miR-196a in both cancer and cancer stroma cells. Finally, inhibition of miR-196a counteracted cancer cell proliferation in both laryngeal cancer-derived cells and mouse xenograft model. Conclusions/Significance: Our study provided the possibilities that miR-196a might be very useful in diagnosing and treating laryngeal cancer. To explore laryngeal cancer-specific miRNAs, RNA from 5 laryngeal surgical specimens including cancer and non-cancer tissues were hybridized to microarray carrying 723 human miRNAs. Total RNA including low molecular weight RNA from tissue samples was isolated using the mirVanaTM miRNA Isolation Kit (Ambion) according to the manufacturer's instructions. The quality of the RNA samples was assessed using an Agilent 2100 Bioanalyzer, and only the samples meeting the criteria of 28S/18S > 1 and RNA Integrity Number (RIN) ≥ 7.5 were used for all analyses. For microarray analysis, we used the Human miRNA Microarray Kit V2 (Agilent Technologies, Santa Clara, CA), which contains 20-40 features targeting each of 723 human miRNAs (Agilent design ID 019118) as annotated in the Sanger miRBase, release 10.1. Labeling and hybridization of total RNA samples were performed according to the manufacturer's protocol. One hundred ng of total RNA was used as an input into the labeling reaction, and the entire reaction was hybridized to each array for 20 hours at 55°C. The results were analyzed using Agilent GeneSpring GX7.3. Normal controls and cancer samples were compared using Welch's t-test (p<0.05) and differentially expressed miRNAs with at least a 2-fold change in expression were considered to be potential biomarkers.
Project description:TGFβ activates a signal transduction cascade that results in the microRNAs and genes transcription. The objective of this study is to identify miRNAs which are regulated through TGFB signaling pathway Cultured NHLF cells at 30-50% confluence were treated with 5 ng/mL TGFB1 or DMSO.
Project description:We performed microRNA microarray analysis to determine microRNAs that are differentially expressed between the differentiated hfRPE monolayer (pigmented, hexagonally packed) and dedifferentiated hfRPE cells (non-pigmented, fibroblast-like). Experiment was performed using the Agilent-019118 Human miRNA Microarray 2.0 G4470B. 2 -condition experiment: differentiated and dedifferentiated hfRPE cells. 3 biological replicates for each condition. Also includes a sample that has characteristics of both cell types (representing a transitional state).
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:In order to investigate miRNA alterations associated to early relapse in ovarian cancer patients, we analyzed miRNA expression profile in a training set of 55 surgical specimens including 30 early relapsing and 25 late relapsing patients. Patients were selected on the basis of residual disease after primary surgery and time to relapse (TTR) after front-line chemotherapy. For training set, a selection of the outliers concerning TTR was made: as early relapsing were defined optimally debulked (OD) patients with a TTR< 12 months and sub-optimally debulked (SOD) patients with TTR< 6 months; late relapsing were defined OD patients with TTR> 36 months and SOD patients with TTR> 12 months. Clinical codes: Histotype: according to International Federation of Gynecological and Obstetrics guidelines Stage: according to International Federation of Gynecological and Obstetrics guidelines Grading: according to International Federation of Gynecological and Obstetrics guidelines Debulking: NED: not evident disease; mRD: minimal residual disease; GRD: gross residual disease Therapy code: P: Platinum without taxanes; PT: Platinum/paclitaxel End Point: Early: relapse whithin 12 and 6 months from the end of therapy for optimally (NED+ mRD) and sub-optimally (GRD) debulked patients respectively. Late: median time to relapse 48 and 34 months from the end of therapy for optimally and sub-optimally debulked patients respectively.