Effects of SUV39H1 depletion on gene expression in HeLa cells
ABSTRACT: HeLa cells were transiently transfected with siRNA against SUV39H1, a histone H3K9 methyltransferase. Several genes were significantly up- or down-regulated. HeLa cells were transiently transfected with siRNA against SUV39H1 or negative control siRNA. Then total RNA was extracted.
Project description:The objective of the present study is to investigate the role of processing glycosidase inhibition induced by processing glycosidase inhibitors, namely castanospermin as the processing glucosidases inhibitor, deoxymannojirimycin as the processing mannosidases inhibitor, and deoxyfuconojirimycin as the fucosidase inhibitor. Comparative gene expression analysis was performed in human cervical carcinoma HeLa cells, with non-inhibitor-treated cells as the negative control. Gene expression was analyzed using 3D-Gene® Human 25k oligonucleotide microarrays and computational gene expression analysis tools. Inhibition of processing glycosidases (glucosidases, mannosidases, and fucosidase) in human cervical carcinoma HeLa cells was performed by treating with a small molecule inhibitor and then culturing at 37°C. Total RNA samples were prepared from the cells. Gene expression was analyzed using 3D-Gene® Human 25k oligonucleotide microarrays. Sample preparation for array hybridization was carried out as described in the manufacturer's instructions.
Project description:microRNA profiling of comparing control or ETO2 siRNA-treated human K562 cells Two-condition experiment, K562-control siRNA vs. K562-ETO2 siRNA, Biological replicates: 1, 1 control siRNA, 1 ETO2 siRNA, independently.
Project description:We used microarray to determine the genes whose expression was changed at six hours after doxycycline treatment Cells were transfected with β-globin let7 wt constructs. Two-condition experiment; HeLa tet off cells overexpressed control vs HeLa tet off cells treated with doxycycline for six hours
Project description:Transcriptional profiling of human hTERT-RPE1 cell spheroids comparing Control siRNA transfected hTERT-RPE1 cell spheroids with those transfected with YAP1 siRNA. Two-condition experiment, Control siRNA vs.YAP1 siRNA hTERT-RPE1 cell spheroids. Biological replicates: 1 Control siRNA, 1YAP1 siRNA transfected, independently grown and harvested. Bothreplicates per array.
Project description:Class Switch Recombination (CSR) is a B cell specific genomic alteration induced by activation induced cytidine deaminase (AID)-dependent DNA break, followed by repair and recombination at the immunoglobulin heavy-chain locus. The involvement of several chromatin-associated factors in promoting AID-induced DNA break formation has been reported. However, the involvement of chromatin adaptors at the repair phase of CSR remains unknown. Here, we provide evidence that acetylated histone reader Brd4 is critical to the repair and recombination step of CSR. Brd4 was recruited to the AID-induced DNA break region, and depletion of Brd4 from the S region chromatin by knockdown or a chemical inhibitor JQ1 causes CSR impairment without affecting AID-induced DNA break generation. Such inhibition of Brd4 suppressed the accumulation of 53BP1 and UNG at the cleaved S regions, perturbed switch donor-switch acceptor microhomology length and reduced Igh/c-myc translocation. We conclude that Brd4 serves as a histone-reader platform required for the recruitment of CSR repair components. Brd4 were depleted from the chromatin by either siRNA treatment or JQ1 (40nM) addition in CH12F3-2A cells in the presence of CIT stimulation. RNA from each samples were extracted and relative difference in transcript level were compared with control RNAi- and DMSO-treated, CIT-stimulated samples.
Project description:To identify molecular biomarkers that are useful for diagnosis and its targeting treatment, we analysed expression profile of synovial sarcoma tissue. In the present study, we studied gene expression profiles comparing 11 cases of synovial sarcoma.
Project description:To identify molecular biomakers that are useful for diagnosis and its targeting treatment, we compared the gene expression profile of myxiod liposarcoma with that of normal fat tissue. In the present study, we studied about gene expression profiles comparing 6 non-preoperative myxoid liposarcoma with 3 normal fat tissue.
Project description:We compared the profile of miRNAs expressed in HEK293 and MRC5 cells that overexpressed KRASG12V to those expressed in parental cells that harbored wild-type KRAS. The results indicated that a subset of miRNAs was significantly down-regulated in KRASG12V transfected two cells. To address the functional relevance of miRNAs in KRAS mutant cancers, we transfected exogenous KRASG12V into HEK293 and MRC5 cells with wild type KRAS genes, and we comprehensively profiled the dysregulated miRNAs.
Project description:To elucidate the aetiology of HEH, especially with regard to microRNA (miRNA) profiles, samples obtained from three different parts of both normal and tumour tissues were analysed . The details of RNA extraction have been described in our previous reports. Interestingly, 16 miRNAs were significantly up-regulated and 51 miRNAs were down-regulated in the tumour tissues compared to the normal tissues (Student’s t-test, p<0.01). In addition, unsupervised hierarchical clustering analysis using a Pearson’s correlation showed that the tumour tissues clustered separately from the normal tissues. microRNA samples obtained from three different parts of both normal and tumour tissues were analysed
Project description:Recent studies have demonstrated that micro (mi)RNA molecules can be detected in the circulation and can serve as potential biomarkers of various diseases. This study used microarray analysis to identify aberrantly expressed circulating miRNAs in patients with type 1 autoimmune hepatitis (AIH) compared with healthy controls. Patients with well-documented and untreated AIH were selected from the National Hospital Organization (NHO)-AIH-liver-network database. They underwent blood sampling and liver biopsy with inflammation grading and fibrosis staging before receiving treatment. To further confirm the microarray data, circulating expression levels of miR-21 and miR-122 were quantified by real-time quantitative polymerase chain reaction in 46 AIH patients, 40 patients with chronic hepatitis C (CHC), and 15 healthy controls. Consistent with the microarray data, serum levels of miR-21 were significantly elevated in AIH patients compared with CHC patients and healthy controls. miR-21 and miR-122 serum levels correlated with alanine aminotransferase levels. Circulating levels of miR-21 and miR-122 were significantly reduced in AIH patients with liver cirrhosis, and were inversely correlated with increased stages of fibrosis. By contrast, levels of circulating miR-21 showed a significant correlation with the histological grades of inflammation in AIH. We postulate that aberrantly expressed serum miRNAs are potential biomarkers of AIH and could be implicated in AIH pathogenesis. Alternations of miR-21 and miR-122 serum levels could reflect their putative roles in the mediation of inflammatory processes in AIH. Case-control study, steroid treatment