title: Molecular determinants of human uveal melanoma invasion and metastasis.
abstract: The molecular analysis of cancer has benefited tremendously from the sequencing of the human genome integrated with the science of bioinformatics. Microarray analysis technology has the potential to classify tumors based on the differential expression of genes. In the current study, a collaborative, multidisciplinary approach was utilized to study the molecular determinants of human uveal melanoma invasion and metastasis. Uveal melanoma is considered the most common primary intraocular cancer in adults, resulting in the death of approximately 50% of patients affected. Unfortunately, at the time of diagnosis, many patients already harbor microscopic metastases, thus underscoring a critical need to identify prognostic markers indicative of metastatic potential. The investigative strategy consisted of isolating highly invasive vs. poorly invasive uveal melanoma cells from a heterogeneous tumor derived from cells that had metastasized from the eye to the liver. The heterogeneous tissue explant MUM-2 led to the derivation of two clonal cell lines: MUM-2B and MUM-2C. Further morphological and functional analyses revealed that the MUM-2B cells were epithelioid, interconverted (expressing mesenchymal and epithelial phenotypes) highly invasive, and demonstrated vasculogenic mimicry. The MUM-2C cells were spindle-like, expressed only a vimentin mesenchymal phenotype, poorly invasive, and were incapable of vasculogenic mimicry. The molecular analysis of the MUM-2B vs. the MUM-2C clones resulted in the differential expression of 210 known genes. Overall, the molecular signature of the MUM-2B cells resembled that of multiple phenotypes--similar to a pluripotent, embryonic-like genotype. Validation of select genes that were upregulated and down-regulated was conducted by semiquantitative RT-PCR measurement. This study provides a molecular profile that will hopefully lead to the development of new molecular targets for therapeutic intervention and possible diagnostic markers to predict the clinical outcome of patients with uveal melanoma.
authors.affiliation: Department of Anatomy and Cell Biology, College of Medicine and The Holden Comprehensive Cancer Center, University of Iowa, Iowa City 52242-1109, USA.
authors.forename: Elisabeth A
authors.initials: EA
authors.lastname: Seftor
authors.forename: Paul S
authors.initials: PS
authors.lastname: Meltzer
authors.forename: Dawn A
authors.initials: DA
authors.lastname: Kirschmann
authors.forename: Jacob
authors.initials: J
authors.lastname: Pe'er
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Maniotis
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Trent
authors.forename: Robert
authors.initials: R
authors.lastname: Folberg
authors.forename: Mary J C
authors.initials: MJ
authors.lastname: Hendrix
mesh_terms: D000328:Adult; D015536:Down-Regulation; D005838:Genotype; D006801:Humans; D007150:Immunohistochemistry; D053533:Keratin-8; D007633:Keratins / Q000096:biosynthesis; D008297:Male; D008545:Melanoma / Q000235:genetics* / Q000473:pathology*; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D020411:Oligonucleotide Array Sequence Analysis; D011379:Prognosis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D014604:Uveal Neoplasms / Q000235:genetics* / Q000473:pathology*; D014746:Vimentin / Q000096:biosynthesis
chemical_list: C426203:KRT8 protein, human; D053533:Keratin-8; D014746:Vimentin; D007633:Keratins