title: Conditional activation of RET/PTC3 and BRAFV600E in thyroid cells is associated with gene expression profiles that predict a preferential role of BRAF in extracellular matrix remodeling.
abstract: Papillary thyroid cancers (PTC) are associated with nonoverlapping mutations of genes coding for mitogen-activated protein kinase signaling effectors (i.e., the TK receptors RET or NTRK and the signaling proteins RAS and BRAF). We examined the pattern of gene expression after activation of these oncoproteins in thyroid PCCL3 cells, with the goal of identifying pathways or gene subsets that may account for the phenotypic differences observed in human cancers. We hybridized cDNA from cells treated with or without doxycycline to induce expression of BRAF(V600E), RET/PTC3, or RET/PTC3 with small interfering RNA-mediated knockdown of BRAF, respectively, to slides arrayed with a rat 70-mer oligonucleotide library consisting of 27,342 oligos. Among the RET/PTC3-induced genes, 2,552 did not require BRAF as they were similarly regulated by RET/PTC3 with or without BRAF knockdown and not by expression of BRAF(V600E). Immune response and IFN-related genes were highly represented in this group. About 24% of RET/PTC3-regulated genes were BRAF dependent, as they were similarly modified by RET/PTC3 and BRAF(V600E) but not in cells expressing RET/PTC3 with knockdown of BRAF. A gene cluster coding for components of the mitochondrial electron transport chain pathway was down-regulated in this group, potentially altering regulation of cell viability. Metalloproteinases were also preferentially induced by BRAF, particularly matrix metalloproteinase 3 (MMP3), MMP9, and MMP13. Accordingly, conditional expression of BRAF was associated with markedly increased invasion into Matrigel compared with cells expressing RET/PTC3. The preferential induction of MMPs by BRAF could explain in part the more invasive behavior of thyroid cancers with BRAF mutations.
authors.affiliation: Division of Endocrinology and Metabolism, University of Cincinnati College of Medicine, 3125 Eden Avenue, Cincinnati, OH 45267, USA.
authors.forename: Cleo
authors.initials: C
authors.lastname: Mesa
authors.forename: Mana
authors.initials: M
authors.lastname: Mirza
authors.forename: Norisato
authors.initials: N
authors.lastname: Mitsutake
authors.forename: Maureen
authors.initials: M
authors.lastname: Sartor
authors.forename: Mario
authors.initials: M
authors.lastname: Medvedovic
authors.forename: Craig
authors.initials: C
authors.lastname: Tomlinson
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Knauf
authors.forename: Georg F
authors.initials: GF
authors.lastname: Weber
authors.forename: James A
authors.initials: JA
authors.lastname: Fagin
mesh_terms: D000818:Animals; D002291:Carcinoma, Papillary / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D016000:Cluster Analysis; D004318:Doxycycline / Q000494:pharmacology; D005109:Extracellular Matrix; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D020782:Matrix Metalloproteinases / Q000096:biosynthesis / Q000235:genetics; D048493:Proto-Oncogene Proteins B-raf / Q000096:biosynthesis / Q000235:genetics*; D051096:Proto-Oncogene Proteins c-ret / Q000096:biosynthesis / Q000235:genetics*; D051381:Rats; D013964:Thyroid Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
chemical_list: D007527:Isoenzymes; D051096:Proto-Oncogene Proteins c-ret; C495001:Ret protein, rat; C482123:Braf protein, rat; D048493:Proto-Oncogene Proteins B-raf; D020782:Matrix Metalloproteinases; D004318:Doxycycline