Cyclooxygenase-2 overexpression in human basal cell carcinoma cell line increases antiapoptosis, angiogenesis, and tumorigenesis
Journal
Journal of Investigative Dermatology
Journal Volume
126
Journal Issue
5
Pages
1143-1151
Date Issued
2006
Author(s)
Abstract
Cyclooxygenase-2 (COX-2) is critical for tumor formation, angiogenesis, metastasis, and prognosis. In this study, the role of COX-2 in antiapoptosis, tumorigenesis, and angiogenesis of human basal cell carcinoma (BCC) cells was investigated. Transfection of COX-2 constitutive expression vector into a BCC cell line yielded several overexpressing clones. All transfectants demonstrated remarkable resistance to ultraviolet B-induced apoptosis (confirmed by flow cytometry analysis, morphological change, and DNA fragmentation). Immunoblot analysis revealed marked increases in apoptosis-regulated genes Mcl-1 and Bcl-2. A 10-fold concentrated conditioned medium from COX-2-overexpressing BCC cells exhibited higher angiogenic activity in Matrigel plug and human umbilical vein endothelial cell tube formation assay. Cells exhibited increased levels of vascular endothelial growth factor-A (VEGF-A) mRNA and protein, and secreted VEGF-A and basic fibroblast growth factor (bFGF). COX-2-specific small interfering RNA markedly reduced the secreted species. After 7 weeks of inoculation, the tumor volume of COX-2-overexpressing cells in severe combined immunodeficient mice was significantly greater than that of vector control cells. Immunohistochemical analysis of CD31-positive vessels revealed a two-fold increase in microvessel density in COX-2 tumors, compared to control vector tumors. Our data indicate that Mcl-1 and Bcl-2, as well as VEGF-A and bFGF, are downstream effectors of COX-2-induced antiapoptosis and angiogenesis, respectively. ? 2006 The Society for Investigative Dermatology.
SDGs
Other Subjects
basic fibroblast growth factor; CD31 antigen; cyclooxygenase 2; matrigel; messenger RNA; protein bcl 2; protein mcl 1; small interfering RNA; vasculotropin A; angiogenesis; animal experiment; animal model; animal tissue; apoptosis; article; assay; basal cell carcinoma; cancer cell culture; carcinogenesis; cell structure; controlled study; DNA strand breakage; expression vector; flow cytometry; gene overexpression; genetic transfection; human; human cell; immunohistochemistry; metastasis; microvasculature; mouse; nonhuman; priority journal; prognosis; protein expression; protein secretion; SCID mouse; signal transduction; ultraviolet B radiation; Animals; Apoptosis; Carcinoma, Basal Cell; Cell Line, Tumor; Cyclooxygenase 2; Humans; Mice; Mice, SCID; Neovascularization, Pathologic; RNA, Messenger
Type
journal article