Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Long-noncoding RNAs in basal cell carcinoma

ABSTRACT: A total of 3 patients with basal cell carcinoma (BCC) and 3 healthy individuals (control; non-lesional skin) were enrolled in the study. Punch biopsies (4 mm) were obtained under local anaesthesia and immediately put in RNAlater (Qiagen, Hilden, Germany) and stored at - 80 Â°C until RNA extraction. The lncRNA and mRNA expression in BCC was compared to lncRNA and mRNA expression in non-lesional skin (control).

ORGANISM(S): Homo sapiens

SUBMITTER: Michael Sand

PROVIDER: E-GEOD-74858 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Long-noncoding RNAs in basal cell carcinoma.

Sand Michael M Bechara Falk G FG Sand Daniel D Gambichler Thilo T Hahn Stephan A SA Bromba Michael M Stockfleth Eggert E Hessam Schapoor S

Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 20160209 8

Long noncoding RNAs (lncRNAs) are fundamental regulators of pre- and post-transcriptional gene regulation. Over 35,000 different lncRNAs have been described with some of them being involved in cancer formation. The present study was initiated to describe differentially expressed lncRNAs in basal cell carcinoma (BCC). Patients with BCC (n = 6) were included in this study. Punch biopsies were harvested from the tumor center and nonlesional epidermal skin (NLES, control, n = 6). Microarray-based ln ...[more]

PMID: 26861560

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Project description:Perturbations in the expression profiles of microRNAs (miRNAs) have been reported for a variety of different cancers. Basal cell carcinoma (BCC) of the skin has not been systematically evaluated regarding differentially expressed miRNAs. Patients with BCC (n=7) were included in the study. Punch biopsies were harvested from the center of the tumors (lesional, n=7) and from sites of adjacent non-lesional skin (intraindividual control, n=7). Microarray miRNA expression profiles were detected based on an Agilent platform and miRBase 16. For validation purposes the expression of a subset of dysregulated miRNAs were measured by means of TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR). Seven patients with basal cell carcinoma (BCC) were enrolled in the present study. While BCCs were excised with cold steel under local anaesthesia, a 4-mm punch biopsy was taken from the center of the tumor and from adjacent non-lesional epithelial skin (control), immediately stored in RNAlater (Qiagen, HIlden, Germany) and stored at -80 M-BM-0C. miRNAs were isolated with miRNeasy Mini Kit (Quiagen, Hilden, Germany) according to the manufacturerM-bM-^@M-^Ys protocol. RNA concentration, purity and RNA integrity number (RIN) were determined with NanoDrop ND-1000 spectral photometer (Peqlab, Erlangen, Germany), Agilent 2100 Bioanalyzer and the RNA 6000 NanoLabCHip Kits (both Agilent Technologies, Santa Clara, USA). total RNA samples were spiked with the MicroRNA Spike-In Kit (Agilent Technologies, Santa Clara, USA) for assessment of labeling and hybridization efficiencies. After the spiked total RNA was treated with alkaline calf intestine phosphatase (CIP), a labeling reaction was started with 100 ng total-RNA per sample. T4 RNA ligase, incorporating Cyanine 3-Cytidine biphosphate (miRNA Complete Labeling and Hyb Kit, Agilent Technologies, Santa Clara, USA) was used to label the dephosphorylated RNA. Cyanine-3-labeled miRNA samples were subsequently prepared for One-Color based hybridization (Complete miRNA Labeling and Hyb Kit, Agilent Technologies, Santa Clara, USA). Hybridization of the labeled miRNA samples with Human miRNA Microarrays Release 16.0 (Agilent Technologies, Santa Clara, USA) was done at 55M-BM-0C for 20 hrs. After washing microarray slides with increasing stringency (Gene Expression Wash Buffers, Agilent Technologies, Santa Clara) they were dried with acetonitrile (Sigma-Aldich, St.Louis, USA). Agilent DNA Microarray Scanner (Agilent Technologies, Santa Clara, USA) was used detect fluorescent signal intensities. They were detected with the Scan Control A.8.4.1 Software (Agilent Technologies, Santa Clara, USA) and extracted from images by using the Feature Extraction 10.7.3.1 Software (Agilent Technologies, Santa Clara, USA). All the steps described were carried out according to the manufacturerM-BM-4s instructions.

Project description:Sheep provide considerable materials for the animal fibre industry. Identifying genes of major effect for wool growth would offer strategies for improving the quality and increasing the yield of fine wool. In this study, we employed Agilent Sheep Gene Expression Microarray and proteomic technology to investigate the gene expression patterns of body side skin (more wool growing) in Aohan fine wool sheep (a Chinese indigenous breed) in comparison with groin skin (no wool growing) at the anagen stage of wool follicle. Microarray study revealed that 4772 probes were differentially expressed, including 2071 upregulated and 2701 downregulated probes in the comparisons of body side skin versus groin skin (S/G). The microarray results were verified by means of quantitative PCR. 1099 probes were assigned to unique genes/transcripts. The number of distinct genes/transcripts (annotated) was 926, of which 352 were up-regulated and 574 were down-regulated. In S/G, 13 genes were up-regulated by more than 10-fold, while 60 genes were down-regulated by more than 10-fold. Further analysis revealed that the majority of the genes possibly related to the wool growth could be assigned into the categories including regulation of cell division, intermediate filament, cytoskeletal part and growth factor activity. Several potential gene families might participate in hair growth regulation, including fibroblast growth factors, transforming growth factor-Î², WNTs, insulin-like growth factor, vascular endothelial growth factors and so on. Proteomic analysis also got 196 differentially expressed protein points, of which 121 were identified as single protein points. Furthermore, according to the results at both mRNA and protein levels, similar regulation mechanism of gene activity might be engaged during skin development and embryo development. One ram and two ewes of 12-month-old Aohan fine wool sheep were used in the microarray study. These animals were half sibs (sharing the same father). In August 2010, two areas of full-thickness skin were sampled from the same animal under local anaesthesia: body side skin (wool bearing) and groin skin (non-wool bearing) for microarray and proteomic experiments. The area of each sample was about 1 cm2. All samples were immediately put into collection tubes and stored in liquid nitrogen for RNA and protein extraction. A total of 15, 208 probes were spotted on this Agilent Sheep Gene Expression Microarray (Santa Clara, CA, USA).

Dataset Information

Long-noncoding RNAs in basal cell carcinoma

Publications

Long-noncoding RNAs in basal cell carcinoma.

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