RNA sequencing (RNA-seq) of LAIR-1 overexpression in SKOV-3 ovarian cancer cell lines
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1032090 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR26525704_1.fastq.gz | Fastqsanger.gz | |||
| SRR26525704_2.fastq.gz | Fastqsanger.gz | |||
| SRR26525705_1.fastq.gz | Fastqsanger.gz | |||
| SRR26525705_2.fastq.gz | Fastqsanger.gz | |||
| SRR26525706_1.fastq.gz | Fastqsanger.gz |
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Project description:Leukocyte-associated immunoglobulin-like receptor 1 (LAIR-1), is widely expressed by solid tumor cells and functions as a tumor suppressor. Our previous studies have found that LAIR-1 is expressed in ovarian cancer tumor tissues and in a variety of epithelial ovarian cancer cell lines and that LAIR-1 could inhibit ovarian cancer cell growth by suppressing the PI3K-AKT-mTOR pathway. The aim of the present study was to obtain a comprehensive understanding of the molecular mechanism of LAIR-1 in regulating the biological behavior of ovarian cancer cells. The approach taken was to perform RNA-seq sequencing on SKOV3 cells overexpressing LAIR-1, and to conduct further validation by combinatorial integration of bioinformatics analyses to assess the genetic alterations and using biological experimental findings.
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Project description:We studied the variations of mRNA amounts after Evi1 knockdown or Flag-Evi1 overexpression in SKOV-3 cells. Despites Evi1 discovery in 1988, its recognized role as a dominant oncogene in myeloid leukemia and more recently in epithelial cancers, only a few target genes were known and it was not clear why Evi1 was involved in cancer progression. Here we obtained the genomic binding occupancy and expression data for Evi1 in human ovarian carcinoma cells. We identified numerous Evi1 target cancer genes and genes controlling cell migration and adhesion. Moreover, we characterized a transcriptional cooperation between AP1 and Evi1 that regulated proliferation and adhesion through a feed-forward loop. Furthermore, this study provides human genome-wide mapping and downstream analyses for Evi1 that will be useful for the research community.
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Project description:We studied the variations of mRNA amounts after Evi1 knockdown or Flag-Evi1 overexpression in SKOV-3 cells. Despites Evi1 discovery in 1988, its recognized role as a dominant oncogene in myeloid leukemia and more recently in epithelial cancers, only a few target genes were known and it was not clear why Evi1 was involved in cancer progression. Here we obtained the genomic binding occupancy and expression data for Evi1 in human ovarian carcinoma cells. We identified numerous Evi1 target cancer genes and genes controlling cell migration and adhesion. Moreover, we characterized a transcriptional cooperation between AP1 and Evi1 that regulated proliferation and adhesion through a feed-forward loop. Furthermore, this study provides human genome-wide mapping and downstream analyses for Evi1 that will be useful for the research community. 16 samples were collected. Each condition was done in 4 replicates, collected 65 hours after transfection. Transfections with control siRNA or Flag-expressing vector were used as controls.
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Project description:We intraperitoneally injected SKOV3 (derived from human ovarian adenocarcinoma) expressing soluble fragment of human EP2 receptor or hIgG Fc (SKOV/ip-FuEP2/Ex2 and (SKOV/ip-Fumock). After 4 week, resulted tumors were lysed and total RNAs were isolated. By using microarray, We analyzed gene expression and identified distinct classes of up-regulated genes in SKOV/ip-FuEP2/Ex2-derived tumor.
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Project description:Human paclitaxel-resistant (SKOV-3TR) ovarian cancer cell line and the parent SKOV-3 cell line were grown in RPMI medium containing 10% fetal calf serum. The experiment was done to compare the expression differences between the parent SKOV-3 cell line and the derived SKOV-3TR cell line. Cells were plated in T75 flasks. They were cultured until they were 80% confluent and harvested by trypsinization followed by pelleting of the cells. All RNA isolation was accomplished with the Tri-Reagent (SIgma) and purified using the Qiagen RNeasy Mini Kit reagents. The RNA (50 ug) was annealed with a random hexamer primer, and reverse-transcribed into cDNA with Powerscript (Clontech) reverse transcriptase for 2h at 42 degrees in the presence of amino-allyl dUTP. The cDNA from SKOV-3 RNA and SKOV-3TR RNA was covalently coupled separately with Cy3 and Cy5 monoreactive fluors in 50 mM sodium bicarbonate, pH 9.0. The Cy3 and Cy5 labelled cDNAs were combined and purified with a QIAquick PCR purification kit and hybridized to the microarray using the Hybrite chamber (Vysis). Fluorescent array images were collected for both Cy3 and Cy5 with a ScanArray 4000 (Perkin Elmer) at a resolution of 10 um at maximum laser power and photo-multiplier tube voltage of 50 to 60%. Segmentation was performed using the histogram method of the QuantArray (Perkin Elmer) package. Data was transformed into log base 2 expression ratios and normalized using scaled loess normalization. Keywords: repeat sample
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Project description:A microarray analysis was performed to compare the global gene expression profile between C-CPE treated- and untreated- SKOV-3 ovarian cancer cells. SKOV-3 cells were treated with or without C-CPE for 72 hours, and total RNA was extracted and microarray was perfomed to compare the gene profiling changes between C-CPE treated- and untreated- cells. The experiment was performed in triplicate.
2010-06-23 | E-GEOD-22421 | biostudies-arrayexpress
Project description:Human paclitaxel-resistant (SKOV-3TR) ovarian cancer cell line and the parent SKOV-3 cell line were grown in RPMI medium containing 10% fetal calf serum. The experiment was done to compare the expression differences between the parent SKOV-3 cell line and the derived SKOV-3TR cell line. Cells were plated in T75 flasks. They were cultured until they were 80% confluent and harvested by trypsinization followed by pelleting of the cells. All RNA isolation was accomplished with the Tri-Reagent (SIgma) and purified using the Qiagen RNeasy Mini Kit reagents. The RNA (50 ug) was annealed with a random hexamer primer, and reverse-transcribed into cDNA with Powerscript (Clontech) reverse transcriptase for 2h at 42 degrees in the presence of amino-allyl dUTP. The cDNA from SKOV-3 RNA and SKOV-3TR RNA was covalently coupled separately with Cy3 and Cy5 monoreactive fluors in 50 mM sodium bicarbonate, pH 9.0. The Cy3 and Cy5 labelled cDNAs were combined and purified with a QIAquick PCR purification kit and hybridized to the microarray using the Hybrite chamber (Vysis). Fluorescent array images were collected for both Cy3 and Cy5 with a ScanArray 4000 (Perkin Elmer) at a resolution of 10 um at maximum laser power and photo-multiplier tube voltage of 50 to 60%. Segmentation was performed using the histogram method of the QuantArray (Perkin Elmer) package. Data was transformed into log base 2 expression ratios and normalized using scaled loess normalization.
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