MiRNA profiling from C13K and OV2008 ovarian cancer cell lines
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ABSTRACT:
PROVIDER: PRJNA492314 | ENA |
REPOSITORIES: ENA
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Project description:MicroRNAs (miRNAs) represent a class of small non-coding RNAs that control gene expression by targeting mRNAs and triggering either translation repression or RNA degradation. Emerging evidence suggests the potential involvement of altered regulation of miRNA in the pathogenesis of cancers, and these genes are thought to function as both tumor suppressors and oncogenes. Using microRNA microarrays, we identify several miRNAs aberrantly expressed in human ovarian cancer tissues and cell lines. miR-221 stands out as a highly elevated miRNA in ovarian cancer, while miR-21 and several members of the let-7 family are found downregulated. Public databases were used to reveal potential targets for the highly differentially expressed miRNAs. In order to experimentally identify transcripts whose stability may be affected by the differentially expressed miRNAs, we transfected precursor miRNAs into human cancer cell lines and used oligonucleotide microarrays to examine changes in the mRNA levels. Keywords: Expression data from various ovarian cancer cell lines transfected with pre-microRNA.
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Project description:We investigated whether levels of serum microRNAs (miRNAs) could discriminate women with high-grade serous ovarian epithelial cancer (SEOC) from age matched healthy volunteers. miRNA expression profiling was performed on 4 SEOC cell lines and normal human ovarian surface epithelial cells (HOSE). miR-182, miR-200a, b and c were highly overexpressed in the SEOC cell lines. miR-103, miR-92a and miR-638 displayed relatively invariant expression and with RNU48, were assessed, as putative serum miRNA normalizers. The 7 miRNA and RNU48 were assessed in serum from SEOC patients (n = 30) and age-matched healthy volunteers (n = 32) by qRT-PCR following pre-amplification. No correlation between age and miRNA or RNU48 levels was observed (age range 30-79 years). miR-103 demonstrated the least variance and was chosen to normalize serum volume adjusted results. miR-200a, b and c were significantly elevated in SEOC serum (P = 0.002; 0.003; 0.0003 respectively) and a multivariate model was the best predictive classifier of SEOC (ROC-AUC = 0.806). A correlation between high miR-200b levels and residual macroscopic disease following cytoreductive surgery was identified (P = 0.006). Our results suggest that serum miR-200a, b and c may have utility as diagnostic biomarkers for SEOC. miRNA expression was arrayed in 1 human surface epithelial cells (HOSE) and 4 ovarian cancer cell lines (OV167, OV202, OVCAR-3, PE01). HOSE cells were run in triplicate (including 1 dye-swap). Ovarian cancer cell lines were run in duplicate (including 1 dye-swap).
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Project description:Although many genes associated with the development of resistance to chemotherapy are known, the mechanisms of their regulation are still poorly understood. One of the ways to regulate gene expression is regulation at mRNA level by small noncoding RNA particles – designated as a micro RNA (miRNA). Changes in miRNA expression were also observed in cell line studyes. Downregulation of miR-31 expression correlated with taxane resistance in ovarian cancer cell lines. In contrast upregulation of miR-98-5p was observed in cisplatin (CIS) -resistant cell lines. Changes in miRNAs expression were also noted in another cancers. miR-195 expression was downregulated in temozolomid-resistant glioma cells and miR-203 was downregulated in prostate cancer cells resistant to doxorubicin (DOX). The use of miRNA microarrays to analyse changes in miRNA gene expression is an effective molecular tool for the discovering new miRNA genes involved in drug resistance processes. The present study shows alterations in the miRNA expression levels in the CIS (W1CR), PAC (W1PR1 and W1PR2), DOX (W1DR), and topotecan (TOP) (W1TR) - resistant variants of W1 sensitive ovarian cancer cell line.
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