Project description:Sensitivity to cisplatin is increased in SKOV-3 cells after transfecting the cells with unphosphorylatable PEA-15 (PEA-15AA) but not in cells transfected with the empty vector and phosphomimetic PEA-15 (PEA-15DD). In order to investigate the regulation of the underlying genes that increased the sensitivity to cisplatin after transfection with PEA-15AA, a small and well-annotated Clariom S gene microarray was performed.
Project description:Proteins immunoprecipitated by Neil3 in 4 conditions
- before Cisplatin treatment with inhibition of proteasome
- before Cisplatin treatment without inhibition of proteasome
- after 3h of Cisplatin treatment with inhibition of proteasome
- after 3h of Cisplatin treatment without inhibition of proteasome
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.
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. Tumor tissues from SKOV/ip-Fumock or SKOV/ip-FuEP2/Ex2 were lysed and total RNAs were extracted. And then we analyzed expression status by Affymetrix microarrays.
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
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.
Project description:Based on a time-course study of cisplatin response in ovarian cancer cells with/without suppression of annexin A11 expression using whole genome oligonucleotide microarrays, we identified a set of differentially expressed genes associated with annexin A11 expression and patterns of gene expressions in response to cisplatin exposure. Keywords: human ovarian cancer cell lines 2008 cells (one ovarian cancer cell line) were transfected with ANXA11_RNAi or control_RNAi for 2 days and then treated with 10 µM cisplatin (Sigma) for 0, 8, 16, 24 hours. Total RNA of 8 samples were prepared for gene expression profiling using the Agilent 44K whole genome oligonucleotide microarrays.
Project description:Identification of target genes of PITX2 homeodomain transcription factor in ovary using human ovarian adenocarcinoma cells, SKOV-3 to determine the transcriptional network of PITX2.
Project description:Chemo-resistance to platinum such as cisplatin is critical in the treatment of ovarian cancer. Recent evidences have linked epithelial-mesenchymal transition (EMT) with the drug resistance as a contributing mechanism. The current study explored the connection between cellular responses to cisplatin with EMT in ovarian cancer. 46 ovarian carcinoma cell lines expression data with and without Cisplatin treatment.