Project description:We implementing a transcriptomic approach to identify global miRNA alterations in chronic iron exposed FTSECs. Chronic iron exposure in fallopian tube secretory epithelial cells (FTSECs) leads to alterations in oncogene expression and functional changes reminescent of early changes in ovarian cancer development (serous epithelial high grade ovarian tumors).
Project description:A novel class of genes has recently been identified as anticancer genes, which upon ectopic overexpression selectively destroy the transformed tumour cells, while leaving the untransformed normal cells unharmed. Examples include TRAIL, PAR4 and Orctl3. We have isolated 16 novel anticancer genes of human origin through a gain of function, forward genetic screen in mammalian cells. Among these, FBLN5 was chosen for further investigation. FBLN5 is a secreted ECM glycoprotein the physiological functions of which remain largely elusive, however it is downregulated in numerous malignancies and Fibln5-/- mice exhibited elastic fibre abnormalities including cutis laxa. When transfected in the normal CV-1 cells and their SV-40 transformed counterpart COS-7 cells, FBLN5 caused extensive cell death in latter and not in the former. In order to investigate such differential effects of FBLN5, we performed transcriptional profiling of COS-7 and CV-1 cells upon FBLN5 transfection where wild type COS-7 and CV-1 cells were used as controls. We conducted exon-level expression profiles of > 540,000 transcripts including coding mRNA, long non-coding RNA, microRNA, novel transcripts and also the alternative splice variants. Briefly, FBLN5 caused downregulation of MYC regulated genes in COS-7 cells while opposite was observed in CV-1 cells. Also, FBLN5 caused upregulation of cell death related genes in COS-7 cells but not in CV-1 cells.
Project description:microRNA array of 4 cell lines: WI-38 primary fibroblasts (“Control”), slow growers (early passage after immortalization, Slow), fast growers (extensive passaging after immortalization) and fast growers transformed by constitutively activated mutant H-RasV12 (“Ras”)
Project description:Methionine sulfoxide reductases catalyze the reduction of MetSO back to the correct Met residue. Previously, the gene of Capsicum annuum methionine sulfoxide reductase B2 was isolated and CaMSRB2-overexpressing tomato shows enhanced growth, which may trigger increased resistance to the pathogens. To assess the role of this enzyme in rice, we generated transgenic lines under the control of the rice Rab21 (responsive to ABA protein 21) promoter with/without Bar marker gene. Several physiological tests such as MV and Fv/Fm, indicators of an oxidative stress-inducing agent and a potential maximal PSII quantum yield, respectively strongly suggested CaMSRB2 confers drought tolerance to rice. Using 3′-tiling microarray covering the whole rice genes, we carried out genome-wide expression analyses with CaMsrB2-transformed rice (Oryza sativa L. cv. ILMI). Rice was grown in port for six weeks and treated with drought by water withholding for two days.
Project description:Human Freshly isolated monocyte (CD14++CD16-) and (CD14+ CD16++) were purified from healthy volunteers' blood, and sorted by FAC. We used Taqman miRNA TLDA arrays to performed miRNA profiling
Project description:Methionine sulfoxide reductases catalyze the reduction of MetSO back to the correct Met residue. Previously, the gene of Capsicum annuum methionine sulfoxide reductase B2 was isolated and CaMSRB2-overexpressing tomato shows enhanced growth, which may trigger increased resistance to the pathogens. To assess the role of this enzyme in rice, we generated transgenic lines under the control of the rice Rab21 (responsive to ABA protein 21) promoter with/without Bar marker gene. Several physiological tests such as MV and Fv/Fm, indicators of an oxidative stress-inducing agent and a potential maximal PSII quantum yield, respectively strongly suggested CaMSRB2 confers drought tolerance to rice. Using 3M-bM-^@M-2-tiling microarray covering the whole rice genes, we carried out genome-wide expression analyses with CaMsrB2-transformed rice (Oryza sativa L. cv. ILMI). Rice was grown in port for six weeks and treated with drought by water withholding for two days. A total of 15 chips were used for the microarray experiment. RNA was extracted from plants just before and 2 days after the duration of water withdrawal for the control and the comparison, respectively. Experiments were performed with three or two biological replicates.
Project description:A series of studies have been published that evaluate the chromosomal copy number changes of different tumor classes using array Comparative Genomic Hybridization (array CGH), however the chromosomal aberrations that distinguish the different tumor classes have not been fully characterized. Therefore, we performed a meta-analysis of different array CGH data sets in an attempt to classify samples tested across different platforms. As opposed to RNA expression a common reference is used in dual channel CGH arrays: normal human DNA, theoretically facilitating cross-platform analysis. To this aim, cell line and primary cancer data sets from three different dual channel array CGH platforms obtained by four different institutes were integrated. The cell line data were used to develop preprocessing methods which performed noise reduction and transformed samples into a common format. The transformed array CGH profiles allowed perfect clustering by cell line, but importantly not by platform or institute. The same preprocessing procedures used for the cell line data were applied to data from 373 primary tumors profiled by array CGH, including controls. Results indicated that there is no apparent feature related to the institute or platform and that array CGH allows for unambiguous cross-platform meta-analysis. Major clusters with common tissue origin were identified. Interestingly, tumors of hematopoietic and mesenchymal origins cluster separately from tumors of epithelial origin. Therefore it can be concluded that chromosomal aberrations of tumors from hematopoietic and mesenchymal origin versus tumors of epithelial origin are distinct, and these differences can be picked up by metaanalysis of array CGH data. This suggests the possibility of prospectively using combined analysis of diverse copy number datasets for cancer subtype classification. Keywords: comparative genomic hybridization, meta-analysis, cancer
Project description:Libraries were made to compare the transcriptome of renin lineage cells (RLCs) from wild-type versus ren knockout zebrafish kidneys. RLCs were FAC sorted from pooled kidneys of ren+/+ or ren-/- zebrafish, which carried ren:RFP and acta2:EGFP reporter genes, allowing the isolation of renin-expressing cells and smooth muscle cells.
Project description:MicroRNA levels in non-transformed BEAS-2B bronchial epithelial cells, two lines of mycoplasma transformed BEAS-2B cells, and A549 lung adenocarcinoma cells were measured. Microarray analyses of 1145 microRNAs in A549 lung adenocarcinoma cells and two other transformed lung cell types relative to BEAS-2B bronchial epithelial cells were performed. 106 miRNAs were down-regulated and 69 miRNAs were up-regulated in all three transformed lines