Project description:Little is known about a relationship between intratumor heterogeneity and drug resistant ability in high grade serous ovarian cancer. Using stem cell cloning technique on high grade ovarian cancer, we have cloned ovarian cancer colonies at high efficiency. The heterogeneity of ovarian cancer is recapitulated in cloned cancer colony library, and Taxol treatment (100 nM 3 hrs) has been conducted on cancer library and obtained drug resistant cancer clones in vitro. Using cloned original cancer colonies and drug resistant cancer colonies, we have studied the effect of intratumor heterogeneity on acquisition of drug resistance. Overall design: We have used surgery samples of high grade serous ovarian cancer and cultured cancer cell colony on 3T3 feeder layer. We used Illumina OmniExpress SNP array to genotype samples from normal fimbria (fimbria clones), cancer clones (N clones), passaging cancer clones (NP clones) and drug resistant cancer clones (TR clones). Fimbria, N and NP clones were cloned from original pooled culture of normal fimbria and ovarian cancer cells. TR clones were cloned from Taxol-treated (four times) ovarian cancer cells. All clones are from two patients and those from patient 2 are called CP30-N clones.
Project description:In this study, we performed miRNA profiles analysis of high-grade serous ovarian carcinoma compared to normal fallopian tube fimbria using microarray (Exiqon, Denmark) to evaluate their potential role in the pathogenesis of uterine leiomyoma. miRNA profiling analysis of the 10 samples including 5 high-grade serous ovarian carcinomas and 5 normal fallopian tube fimbria.
Project description:This SuperSeries is composed of the following subset Series: GSE28720: Gene expression analysis of ovarian tumors from mice with knockout of DICER and PTEN GSE28721: Gene expression analysis of human serious ovarian tumors and fimbria control Refer to individual Series
Project description:The cell of origin of serious ovarian cancer is unknown. To create a mouse model for this lethal cancer and identify early cancer biomarkers, we conditionally deleted both Dicer (essential for microRNA biosynthesis) and Pten (a negative regulator of the PI3K pathway) in the female reproductive tract. Beginning at ~3-5 months, these Dicer/Pten mutant mice develop high-grade serious carcinomas that initiate in the stroma of the fallopian tube through a mesenchymal-to-epithelial transition (MET), subsequently envelop the ovary, and then metastasize throughout the peritoneum, resulting in ascites and 100% lethality by 13 months. The fallopian tube cancers demonstrate upregulation of genes encoding known and novel secreted proteins that are potential biomarkers. This study uncovers a new paradigm for the initiation of high-grade serous ovarian cancer. We generated gene expression profiles of 8 human primary serious tumors, and 2 independent samples of human normal fimbria. We defined genes that were high or low in tumors relative to fimbria, and compared these results with those of the correponding mouse model.
Project description:The biological features of ovarian cancer stem cells (OCSC) remain elusive, mainly because 1) most studies so far have focused on cell lines that recapitulate the human disease only to a limited extend; and 2) because the identification of OCSC has relied on markers inferred from different and unrelated tumor types. Our study has harnessed the intrinsic, stemness-related properties of OCSC to identify and isolate this cell subpopulation from primary cultures freshly established from high-grade serous ovarian cancer (HGSOC), the most common and aggressive from of the disease. In addition, OCSC were compared to stem cell-enriched cultures from fallopian tube epithelium, which is the most accredited tissue of origin for HGSOC. The transcriptomes of the two cell types were compared to infer genes differentially regulated in OCSC. We used affymetrix expression microarray to analyze the transcriptional profile of spheres deriving from high-grade serous ovarian cancer or from normal ovarian fimbria. Overall design: Total RNA was extracted from primary cells derived from either 7 HGSOC ascites samples or 9 normal ovarian fimbriae samples. Samples were pooled together (3-5 samples/pool; Ascites, N=2; Fimbria, N=2) and analyzed using the Affymetrix Human Gene 2.1-st, following manufacturer's standard protocols. Raw affymetrix data were analyzed ausing the Expression Consol suite (Affymetrix) and normalized by RMA_sketch.
Project description:To overview compound-responsive genes in fibroblast, we performed microarray analysis of ~63,000 genes in compound-treated fibroblast of LSND3 Overall design: We treated the fibroblasts with 1μM compound, extracted RNAs from compound- and non-treated fibroblasts at 24 h after treatment, and compared the gene expression ratio between the two groups. Two duplicate experiments.
Project description:Transcriptome profiling using Affymetrix GeneChip arrays was performed on sorted populations of Lgr5-expressing mouse ovarian surface epithelium. The ovary surface epithelium (OSE) undergoes ovulatory tear-and-remodelling throughout life. Resident stem cells drive such tissue homeostasis in many adult epithelia, but their existence in the ovary has yet to be definitively proven. Lgr5 marks stem cells in multiple epithelia. Here we use reporter mice and Single Molecule Florescent-in-Situ-Hybridization (FISH) to document candidate Lgr5+ stem cells within the mouse ovary and associated structures. Lgr5 is broadly expressed during ovary organogenesis, but becomes limited to the OSE in early neonate life. In adults, Lgr5 expression is predominantly restricted to proliferative regions of the OSE and fimbria- mesovarian junction. Using conditional in vivo lineage tracing we identify embryonic and early neonate Lgr5+ populations as stem/progenitor cells contributing to the development of adult OSE and granulosa cell lineages, as well as the epithelia of the mesovarian and oviduct including its distal opening, fimbria. Long-term lineage tracing reveals that adult OSE-resident Lgr5+ populations contribute to epithelial homeostasis and OSE regenerative repair in vivo. Thus, Lgr5 is a marker of stem/progenitor cells of the ovary and tubal epithelia. Ovarian surface epithelium from pooled batches of Lgr5-eGFP-CreERT2 mice (n=8, per array) were sorted for cells expressing either high or low EGFP. Total RNA from three technical replicates per sorted population (Lgr5-high or Lgr5-low) was extracted with the RNeasy Micro Kit (QIAGEN), DNaseI-treated, and amplified with the Ovation Pico WTA V2 (NuGEN Technologies). Single-stranded cDNA amplification products were purified using QIAquick PCR Purification Kit (QIAGEN). cDNA was biotinylated using the Encore Biotin Module (NuGEN Technologies). Biotiylated cDNA was hybridized to Affymetrix Mouse Genechip ST 2.0 expression arrays.
Project description:The virulon of Staphyloccocus aureus is controlled by intricate connections between transcriptional and post-transcriptional regulators including proteins and small non coding RNAs (sRNAs). Most of the sRNAs regulate gene expression through base-pairings with mRNAs. Here, we have determined the transcriptome of RsaI sRNA of S. aureus. Overall design: Global overview of RsaI impact on gene regulation, a comparative transcriptomic analysis was performed on total RNAs extracted from the WT HG001 strain, the isogenic HG001∆rsaI mutant strain, and the same mutant strain complemented with a plasmid expressing RsaI under the control of its own promoter