Expression profiling reveals that methylation of TIMP3 is involved in uveal melanoma development.
Uveal melanoma is associated with a high tumor-related mortality due to the propensity to develop metastases. The mechanisms that are responsible for malignant dissemination are largely unknown and need to be explored to facilitate diagnosis and treatment of metastases. To identify molecules involved in dissemination, we used cell lines derived from a primary uveal melanoma and 2 liver metastases from the same patient as a model for tumor progression. Using a microarray representing 1,176 genes, we identified 63 differentially expressed genes. Forty genes showed a higher expression and 23 showed a lower expression in the primary cell line compared to the metastasis cell lines. These genes are involved in processes like angiogenesis, apoptosis, macrophage stimulation, and extracellular matrix regulation. In contrast, the 2 liver metastasis cell lines produced nearly identical expression profiles. Demethylation of the primary melanoma cell line by 5-aza-2'deoxycytidine treatment revealed that 19 genes were suppressed by hypermethylation. An important finding was the 5-fold decreased expression of TIMP3 in the metastatic cell lines, a molecule involved in extracellular matrix degradation. We demonstrate in the cell lines that TIMP3 expression is regulated by methylation. These observations were confirmed in primary uveal melanoma and suggests a role for TIMP3 in uveal melanoma development.
van der Velden
Pieter A
PA
Department of Dermatology, Leiden University Medical Center, Leiden, The Netherlands.
Zuidervaart
Wieke
W
Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands.
Hurks
Monique H M H
MHMH
Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands.
Pavey
Sandra
S
Queensland Institute of Medical Research, Brisbane, Australia.
Ksander
Bruce R
BR
Schepens Eye Research Institute and the Department of Ophthalmology, Harvard Medical School, Boston, MA, USA.
Krijgsman
Elise
E
Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands.
Frants
Rune R
RR
Center for Human and Clinical Genetics section Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Tensen
Cornelis P
CP
Department of Dermatology, Leiden University Medical Center, Leiden, The Netherlands.
Willemze
Rein
R
Department of Dermatology, Leiden University Medical Center, Leiden, The Netherlands.
Jager
Martine J
MJ
Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands.
Gruis
Nelleke A
NA
Department of Dermatology, Leiden University Medical Center, Leiden, The Netherlands.
D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014408:Biomarkers, Tumor; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors; D015342:DNA Probes; D000077209:Decitabine; D018450:Disease Progression; D015536:Down-Regulation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007124:Immunoenzyme Techniques; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000556:secondary; D008545:Melanoma / Q000201:enzymology / Q000235:genetics* / Q000556:secondary; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000378:metabolism; D012334:RNA, Neoplasm / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019717:Tissue Inhibitor of Metalloproteinase-3; D019714:Tissue Inhibitor of Metalloproteinases / Q000235:genetics*; D014407:Tumor Cells, Cultured; D014604:Uveal Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology
D014408:Biomarkers, Tumor; D015342:DNA Probes; D004791:Enzyme Inhibitors; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; C088390:TIMP3 protein, human; D019717:Tissue Inhibitor of Metalloproteinase-3; D019714:Tissue Inhibitor of Metalloproteinases; D000077209:Decitabine; D015254:DNA Modification Methylases; D001374:Azacitidine