Effects of siRNA targeting PRKCD in breast cancer cells
ABSTRACT: The aim was to identify genes that were commonly influenced by a siRNA targeting PRKCD in breast cancer cell lines. MDA-MB-468 and BT-549 breast cancer cell lines were treated with control siRNA or siRNA targeting PRKCD. Three samples in each group were analyzed.
Project description:Global gene expression profiling was performed on paired tumor biopsies collected before and after 2 weeks of statin treatment with the aim of detecting statin induced changes on tumoral gene expression. In this phase II clinical study using the “window-of-opportunity” design, in which the treatment-free window between a cancer diagnosis and surgical tumor resection is used to study the biological effects of a certain drug, atorvastatin, a lipophilic statin, was prescribed to patients with primary breast cancer for two weeks pre-operatively. Tumor samples subjected to whole genome transcriptional profiling were collected before patients started treatment and after completing treatment.
Project description:Female BALB/c nude mice (n=3/group) were subjected to partial body irradiation under anesthesia using 2 Gy external photon (4 MV, nominal) irradiation. In group A, the collum (i.e. the thyroid) was irradiated; in group B, thorax+abdomen were irradiated, and in group C, collum+thorax+abdomen were irradiated. The control group (n=5) was anesthetized but not irradiated. At 24 h after treatment, the kidneys, liver, lungs, spleen, and thyroid were excised, flash-frozen, and stored at -80°C. Total RNA was extracted from homogenized tissue samples (kidney cortex and kidney medulla were treated separately) and subjected to expression analysis using RNA microarray technology. The study consisted of 3 irradiated groups with 3 animals each and one non-irradiated control group with 5 animals. Six tissues were analyzed per animal, each sample was analyzed individually, i.e. no samples were pooled.
Project description:Circadian rhythm study on transcriptional responses to i.v. administered 90 kBq iodine-131 after 24h in mouse kidney cortex and medulla, liver, lungs, spleen, and thyroid. Female BALB/c nude mice (n=4/group) were i.v. injected with 90 kBq 131I at three different times of day, i.e. at 9.00 am, at 12.00 pm, or at 3.00 pm, and killed under anesthesia after 24h following treatment for excision of organs. For each time of day, a control group (n=3-4/group) was i.v. injected with physiol. saline. The kidneys, liver, lungs, spleen, and thyroid were excised, flash-frozen, and stored at -80°C until extraction of total RNA. Total RNA was extracted from homogenized tissue samples and subjected to expression analysis using RNA microarray technology. The study consisted of 6 groups in total, i.e. 3 groups (n=4/group) that were i.v. injected with 90 kBq 131I (at 9.00 am, 12.00 pm, or 3.00 pm) and 3 sham-treated control groups (n=3-4/group) that were i.v. injected with physiol. saline (at 9.00 am, 12.00 pm, or 3.00 pm); please note that the number of control animals varied between 3-4 animals per group, i.e. n=4 treated at 9.00 am, n=3 treated at 12.00 pm, and n=3 treated at 3.00 pm. Six tissues were analyzed per animal, i.e. kidney cortex, kidney medulla, liver, lungs, spleen, and thyroid.
Project description:RNA microarray analysis of low-dose and dose rate responses versus time after i.v. administration of 211At. Female BALB/c nude mice were intravenously injected with 1.7 kBq 211At and killed after 1 h, 6 h, or 7 d, or injected with 105 or 7.5 kBq and killed after 1 h and 6 h, respectively. Controls were mock-treated. Kidneys, liver, lungs, and spleen were excised, flash-frozen, and stored at -80°C. Total RNA was extracted and subjected to expression analysis using RNA microarray technology. Enriched biological processes were categorized after cellular function based on Gene Ontology terms. Total RNA was isolated from fresh-frozen tissue samples. Three mice per group were used for 1.7 kBq 211At injections and controls; two mice per group for 105 and 7.5 kBq injections.
Project description:Epithelial ovarian cancer is morphologically and clinically heterogeneous. Transcriptional profiling has revealed molecular subtypes (referred to as “C-signatures”) that correlate to biological as well as clinical features. We aimed to determine gene expression differences between malignant, benign and borderline serous ovarian tumors, and to investigate similarities to the intrinsic molecular subtypes of breast cancer. Global gene expression profiling was performed using Illumina's HT12 Bead Arrays and applied to 59 fresh-frozen ovarian tumors. SAM analysis revealed enrichment of cell cycel processes among the malignant tumors, in line with malignant tumors being highly proliferative. The borderline tumors were split between the malignant and benign tumor clusters, indicating that borderline tumors have both malignant and benign features. Furthermore, nearest centroid classification was performed applying previously published gene profiles for the ovarian cancer C-signatures and the intrinsic breast cancer subtypes, respectively, and showed significant correlations between the malignant serous tumors and the highly aggressive C1, C2 and C4 ovarian cancer signatures, and the basal-like breast cancer subtype. The benign and borderline serous tumors together were significantly correlated to the normal-like breast cancer subtype and the ovarian cancer C3 signature. The borderline tumors, on the other hand, correlated significantly to the Luminal A breast cancer subtype. These findings remained when analyzed in a large, independent dataset. The data in this study link the transcriptional profiles of serous ovarian cancer to the intrinsic molecular subtypes of breast cancer, in line with the shared clinical and molecular features between high-grade serous ovarian cancer and basal-like breast cancer, including an aggressive phenotype, frequent TP53 mutations and a high degree of genomic instability, and suggest that biomarkers and targeted therapies may overlap between these subsets of ovarian and breast cancers. Finally, the link between benign and borderline ovarian cancer and luminal breast cancer may indicate endocrine responsiveness in a subset of ovarian cancers. Total RNA obtained from serous ovarian adenocarcinomas, adenomas and borderline tumors. Gene expression profiling using Illumina's HT12 v4 bead arrays. Application of ovarian cancer molecular subtypes and intrinsic breast cancer subtypes using nearest centroid classification. KRAS and BRAF mutation analyses in the malignant and borderline tumors.
Project description:Background: Mesenchymal stromal cells (MSC) have been proposed as a future cell based therapy for lung diseases like bronchiolitis obliterans syndrome (BOS). Theoretically, it might be beneficial to use lung resident MSC already adapted to the pulmonary environment. We therefore aimed to compare lung MSC with the well-characterized bone marrow MSC. Furthermore, MSC isolated from lung-transplanted patients with BOS were compared to MSC isolated from good outcome recipients. Methods: MSC were isolated from bone marrow (BM) and adult/fetal lung tissues and compared by a comprehensive panel of assays. Results:The gene expression profiles of adult lung derived and fetal lung derived MSC were very similar compared to BM derived MSC, with only 235 and 338 genes, respectively, being significantly differently expressed. Out of the 235 genes that were significantly different between adult lung derived and BM derived MSC, 90 genes were higher expressed in the lung derived MSC. In case of fetal lung derived MSC, 166 genes were higher expressed compared to the BM derived MSC. Interestingly, only, 89 genes were found to be expressed differently when comparing BM derived MSC to both, fetal and adult lung derived MSC, indicating that these genes are lung specific. Next, we went on to evaluate if MSC isolated from biopsies of lung-transplanted patients with BOS differed compared to patients without BOS, i.e. good outcome recipients. Here we found that four genes (Sox9, FAR2, LOC728855 and NDUFS5) were significantly higher expressed in MSC isolated from BOS patients Conclusions: This data demonstrates that lung resident MSC possess lung specific properties that should be taken into considerations when using MSC for cell-based therapy in severe lung disorders like BOS. These results also show that MSC isolated from lung-transplanted patients with BOS do not have an altered phenotype. Comparison of gene expression of MSC isolated from bone marrow, adult lung and fetal lung.
Project description:Whole genome expression data on transcriptome of human osteosarcoma (HOS) cells induced by bisfenol A (BPA), S (BPAS) and AF (BPAF) after short- (8 hours) and long-term (3 months) exposure at human-relevant (10 nM) concentrations. 8 experimental conditions, 4x2 design: 3 compounds (bisphenol A, S and AF) and a control, 2 exposure times (8 hours and 3 months). 3 replicates for each condition, 24 arrays in total.
Project description:The fallopian tube transports the gametes to the fertilization site and delivers the embryo to the uterus at the optimal time for implantation. Progesterone and the classical progesterone receptor (PGR) are known to be involved in regulating both tubal ciliary beating and muscular contractions, possibly involving both genomic and non-genomic actions. To provide more clues on the mechanisms involved, we investigated the effect of progesterone on gene expression in mice fallopian tubes in vitro at early (20 min) and later (2 h, 8 h) time-points using microarray and/or quantitative PCR. In parallel, oocyte cumulus complex transport was investigated in ovulating mice injected with one of the PGR antagonists, Org 31710 or CDB2194. Microarray analyses did not reveal any apparently regulated genes 20 min after progesterone treatment, in agreement with a proposed non-genomic action of progesterone controlling ciliary beating. After 2 h, 11 genes were significantly up-regulated. Analyses by quantitative PCR at 2 h and 8 h showed a consistent up-regulation of endothelin 1 (Edn1) and a down-regulation of its receptor Ednra by progesterone. We also show that treatment with progesterone receptor antagonist before ovulation accelerates the transport of the oocyte cumulus complex. This is the first study showing that progesterone regulates Edn1 and Ednra in the fallopian tube. Together with previous studies on endothelin-mediated effects on muscular contractions in the fallopian tube, the results from this study suggest that endothelin is a mediator of the progesterone-controlled effects on muscular contraction, and eventually gamete transport, in the fallopian tube. 16 pooled samples from mice fallopian tubes were exposed in vitro to progesterone for up to 2 hours.
Project description:High and moderate fibrosis inducing carcinoma cell lines gene expression and effect on human fibroblasts in 3 dimensional collagen gels. Cells grown in free floating 3 dimensional collagen gells contining cells for 48 hrs