Circulating U2 small nuclear RNA fragments as a novel diagnostic biomarker for pancreatic and colorectal adenocarcinoma
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ABSTRACT: Global microRNA expression profiling of serum were collected using Agilent miRNA microarrays (G4471A Human, Amadid 29297, Sanger 14) carrying 887 individual human miRNA probes. Two different sources of RNA were analyzed: serum from NMRI-nu/nu mice carrying a xenograft tumor derived from human primary pancreatic ductal adenocarcinomas (CA, n=6) and serum from tumor free control NMRI-nu/nu mice (N, n=6)
Project description:Global microRNA expression profiling of serum were collected using Agilent miRNA microarrays (G4471A Human, Amadid 29297, Sanger 14) carrying 887 individual human miRNA probes. Two different sources of RNA were analyzed: serum from NMRI-nu/nu mice carrying a xenograft tumor derived from human primary pancreatic ductal adenocarcinomas (CA, n=6) and serum from tumor free control NMRI-nu/nu mice (N, n=6) Two condition (CA, N), each condition is represented by 6 biological replicates
Project description:Global microRNA expression profiling of serum were collected using Agilent miRNA microarrays (G4471A Human, Amadid 29297, Sanger 14) carrying 887 individual human miRNA probes. Two different sources of RNA were analyzed: serum from healthy controls (N=5) and patients with ovarian carcinoma (CA, n=5)
Project description:Global microRNA expression profiling of serum were collected using Agilent miRNA microarrays (G4471A Human, Amadid 29297, Sanger 14) carrying 887 individual human miRNA probes. Two different sources of RNA were analyzed: serum from healthy controls (N=5) and patients with ovarian carcinoma (CA, n=5) Two conditions (CA, N), each condition is represented by 5 biological replicates
Project description:Purpose: we aimed at identify and compare the transcriptional changes of ALDH- and ALDH+ DU145 xenografts upon radiotherapy treatment. Method: Xenografts were generated by injection of ALDH- and ALDH+ DU145 cells in male NMRI-Foxn1 nu/nu immune-deficient mice and subjected to fractionated irradiation to a final dose of 50 Gy. Tumors were excised and processed for total RNA extraction and RNAseq analysis.
Project description:To profile lung miRNA expression in our mouse model of cigarette smoke-induced chronic obstructive pulmonary disease, we employed the Agilent unrestricted Mouse miRNA (8 x 15k arrays per slide, AMADID Number: 021828, Sanger Version 12) platform as a discovery tool to identify miRNAs of interest in the development of experimental chronic obstructive pulmonary disease. Mice were exposed to cigarette smoke (or room air) for 4, 6, 8, 12 weeks, lungs were excised, and total RNA isolated.
Project description:Exosomes were purified from 250 ul matched plasma and serum samples using ExoQuickTm. The presence of particles consistent in size with exosomes (60-150nm) was confirmed using a Nanosight LM10. miRNA was extracted from exosomes using an miRNeasy Serum/Plasma kit (Qiagen, #217184). miRNA was reversed transcribed using a TaqMan® microRNA Reverse Transcription Kit (Life technologies, #4366596). miRNA profiling was performed with a high throughput TaqMan® OpenArray® Human microRNA panel (Life technologies, #4461104). The panel consisted of probes for 754 human miRNAs that are based on miRNA sequences derived from Sanger miRBase v14. MegaplexTM Primer Human Pool A v2.1 and Human Pool B v2.0 or v3.0
Project description:A total of 64 NMRI nu/nu male mice (Janvier, Le Genest Saint Isle, France) were divided in four equal groups that underwent subcutaneous implantation of tumor fragments of PC346C, PC2400, PC2416 or PC2416-DEC. Tumor volume was measured twice weekly by calipers. At a tumor volume of 300 mm3, mice were randomized 1 to 1 to receive docetaxel once (33 mg/kg bodyweight, i.v.) and darolutamide q.d. (200 mg/kg bodyweight, p.o.) or both vehicles (5% glucose/EtOH/Polysorbate 80 8:1:1 (v/v/v) i.v. and PEG400/propylene glycol/5% glucose 5:3:2 (v/v/v) p.o., respectively) until tumor volume exceeded 1500 mm3 or for maximal 60 days, at which tumors were harvested and frozen. Drug dosages were based on a previously conducted safety experiment. One mouse was excluded from the analysis due to attrition.
Project description:siRNA mediated gene knockdown has been shown to be extremely sequence specific. However, off-target gene regulation due to partial sequence homology has also been reported. Furthermore, sequence unrelated effects on gene regulation can occur when siRNA is applied in vivo. In this experiment we investigated off target gene regulation due to treatment of mice with siRNA and LNA modified siRNA targeting GFP. Off target gene regulation was observed and was markedly reduced upon introduction of LNA modifiecation in siRNA. Experiment Overall Design: NMRI nu/nu mice were treated with saline (control), siRNA or siLNA for three weeks. Then the mice were sacrificed and tumor total RNA was isolated. The effect of siRNA/siLNA treatment was evaluated by comparing gene expression of saline treated animals with that of siRNA and siLNA treated animals respectively.
Project description:Papillomaviruses (PVs) are able to induce papillomas, premalignant lesions, and carcinomas in a wide variety of species. PVs are classified first based on their host and tissue tropism and then their genomic diversities. A laboratory mouse papillomavirus, MmuPV1 (formerly MusPV), naturally infects NMRI-Foxn1nu/Foxn1nu (nude; T cell deficient) mice. C57BL/6J wild-type mice were not susceptible to MmuPV1 infection; however, immunocompetent, alopecic, S/RV/Cri-ba/ba (bare) mice developed small papillomas at injection sites that regressed. NMRI-Foxn1nu and B6.Cg-Foxn1nu but not NU/J-Foxn1nu mice were susceptible to MmuPV1 infection. B6 congenic strains, but not other congenic strains carrying the same allelic mutations, that lack B- and T-cells, but not B-cells alone, were susceptible to infection, indicating that mouse strain and T-cell deficiency are critical to tumor formation. Although lesions initially observed were exophytic papillomas around the muzzle, exophytic papillomas on the tail and condylomas of the vaginal lining could be induced by experimental infections. On the dorsal skin, locally invasive, poorly differentiated tumors developed with features similar to human trichoblastomas. Transcriptome analysis revealed significant differences between the normal skin in these anatomic sites and in papillomas versus trichoblastomas. The primarily dysregulated genes involved molecular pathways associated with cancer, cellular development, cellular growth and proliferation, cell morphology, and connective tissue development and function. Surprisingly, few of the genes commonly associated with basal cell carcinoma or squamous cells carcinoma were dramatically dysregulated.