Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of human xenografted nasopharyngeal carcinoma tumor cell lines and an untransformed diploid fibroblast cell line

ABSTRACT: The aim of this experiment was to investigate the gene expression profile of two xenografted nasopharyngeal carcinoma tumors - C15 and C17 which are international reference tools for biological investigations of this type of disease. C15 and C17 transcriptome was profiled by comparison with human untransformed diploid fibroblasts, MRC5. These fibroblasts are very sensitive to contact inhibition and were collected after 3 days of confluence, therefore in a quiescent state. We acknowledge that malignant nasopharyngeal carcinoma cells and MRC5 cells do not belong to the same lineage (epithelial cells versus fibroblasts). However this combination was expected to highlight genes involved in proliferation of NPC cells. Competitive hybridisation of cDNA for each pair C15 and MRC5, C17 and MRC5 was performed on Agilent microarrays containing 22 000 60mer oligonucleotides related to 16 000 human transcripts. The analysis was focused mainly on genes overexpressed in both C15 and C17 by comparison with MRC5

ORGANISM(S): Homo sapiens

SUBMITTER: Hugues Ripoche

PROVIDER: E-TABM-382 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Context : Non Functioning Pituitary Adenomas (NFPAs), although typically benign, may be locally invasive. Few datas are currently available related to the molecular process of invasion in sporadic pituitary adenomas. Markers of invasiveness, important for helping the clinician in the therapeutic strategy particularly in the decision for adjuvant radiotherapy, are currently lacking. Objective: Evaluate if invasive NFPAs display a specific expression profile compared with non invasive tumors. Methods: To address this issue, we selected 40 NFPAs (38 of the gonadotroph type) and classified them as invasive (n=22) or non invasive (n=18) on the basis of MRI and surgical findings. Then we performed pangenomic analysis based on Agilent Human Whole genome Gene Expression oligonucleotide microarray (44k) Expression in order to identify genes differentially expressed between invasive and non invasive NFPA. Experiements are made in dual color with tumor samples labelled in cyanine 5 and a pool of all tumors labelled in cyanine 3.The expressions of some genes identified by microarray screening were confirmed by real-time quantitative RT-PCR. The selection of genes was made on the basis of Ingenuity networks and degree of upregulation between invasive and non invasive tumors. Moreover, some genes of interest already described in the literature were added to the analysis. Results: Prediction class analysis showed that 346 genes discriminated between invasive and non invasive NFPAs (p<0.001); 233 were up- and 113 were down-regulated between the two groups. We then tested On the basis of Ingenuity networks and degree of upregulation between invasive and non invasive tumors, a set of eight genes, including MYO5A, IGFBP5, FLT3, NFE2L1, PTTG, MMP9, NCAM and NR1H3, was tested in quantitative PCR analysis and. Results confirmed those obtained in microarrays results.analysis. At the protein level, Myosin 5A (MYO5A) demonstrated stronger immunostaining in invasive NFPAs compared to non invasive NFPAs. Conclusions: We propose a molecular signature of eight genes of grossly invasive NFPAs as compared with non invasive tumors: The product of one of these genes, MYO5A, may be a useful marker of invasive process in tumoral specimens. The role of these genes in the invasiveness process and as prognostic markers of pituitary adenomas needs to be confirmedinvestigated. Method: Microarray analyses the transcriptome of 22 invasive Non Functional Pituitary Adenomas (NFPAs) and 18 non invasive NFPAs in dual color (each sample labelled in cyanine 5 and a pool of all tumors labelled in cyanine 3).

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Transcription profiling of human xenografted nasopharyngeal carcinoma tumor cell lines and an untransformed diploid fibroblast cell line

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