Dominant-negative CREB inhibits tumor growth and metastasis of human melanoma cells.
The ATF/CREB family of eukaryotic transcription factors contain the bZIP structural motif and mediate their transcriptional activities via heterodimerization with ATF and AP-1 family members. Quenching of CREB-associated proteins by a dominant-negative CREB (KCREB) that is mutated within its DNA-binding domain decreases radiation resistance of human melanoma cells. The purpose of this study was to determine the role of CREB in tumor growth and metastasis of human melanoma using KCREB. Highly metastatic MeWo human melanoma cells were transfected with the KCREB expression vector and subsequently analysed for changes in their tumorigenic and metastatic potential. Expression of KCREB in MeWo human cells decreased their tumorigenic and metastatic potential in nude mice compared with parental and control transfected cells. The KCREB-transfected cells displayed downregulation of 72 kDa collagenase type IV (MMP-2) mRNA expression and activity and decreased invasiveness through Matrigel-coated filters. Moreover, transcriptional activities mediated by the CAT gene driven by the MMP-2 promoter were decreased by 14-45-fold in KCREB-transfected cells. In addition, the cell-surface adhesion molecule MCAM/MUC18 that is involved in metastasis of human melanoma was downregulated in the KCREB-transfected cells. These data indicate that, through their transcriptional activities, CREB and its associated proteins play an important role in the acquisition of the metastatic phenotype of human melanoma cells.
Xie
S
S
Department of Cell Biology, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Price
J E
JE
Luca
M
M
Jean
D
D
Ronai
Z
Z
Bar-Eli
M
M
D000818:Animals; D015703:Antigens, CD; D051929:CD146 Antigen; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003094:Collagen; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D004338:Drug Combinations; D018093:Gelatinases / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D006801:Humans; D007797:Laminin; D008175:Lung Neoplasms / Q000556:secondary*; D008297:Male; D020778:Matrix Metalloproteinase 2; D008545:Melanoma / Q000378:metabolism / Q000473:pathology* / Q000556:secondary*; D008562:Membrane Glycoproteins / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D009368:Neoplasm Transplantation; D019006:Neural Cell Adhesion Molecules; D011401:Promoter Regions, Genetic; D011509:Proteoglycans; D014162:Transfection; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D051929:CD146 Antigen; D017362:Cyclic AMP Response Element-Binding Protein; D004338:Drug Combinations; D007797:Laminin; C497902:MCAM protein, human; C497903:Mcam protein, mouse; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D019006:Neural Cell Adhesion Molecules; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D015500:Chloramphenicol O-Acetyltransferase; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2