Heparanase is involved in angiogenesis in esophageal cancer through induction of cyclooxygenase-2.
OBJECTIVE
Both heparanase and cyclooxygenase-2 (COX-2) are thought to play critical roles for tumor malignancy, including angiogenesis, although it is unknown about their relationship with each other in cancer progression. We hypothesized that they may link to each other on tumor angiogenesis.


METHODS
The expressions of heparanase and COX-2 in 77 primary human esophageal cancer tissues were assessed by immunohistochemistry to do statistical analysis for the correlation between their clinicopathologic features, microvessel density, and survival of those clinical cases. Human esophageal cancer cells were transduced with heparanase cDNA and used for reverse transcription-PCR and Western blot to determine the expression of heparanase and COX-2. COX-2 promoter vector and its deletion/mutation constructs were also used along with transduction of heparanase cDNA for luciferase assay.


RESULTS
Heparanase and COX-2 protein expression exhibited a similar pattern in esophageal tumor tissues, and their expression correlated with tumor malignancy and poor survival. Their expression also revealed a significant correlation with high intratumoral microvessel density. Up-regulation of COX-2 mRNA and protein was observed in esophageal cancer cells transfected with heparanase cDNA. COX-2 promoter was activated after heparanase cDNA was transduced and the deletion/mutation of three transcription factor (cyclic AMP response element, nuclear factor-kappaB, and nuclear factor-interleukin-6) binding elements in COX-2 promoter strongly suppressed its activity.


CONCLUSIONS
Our results suggest that heparanase may play a novel role for COX-2-mediated tumor angiogenesis.
Okawa
Takaomi
T
Department of Gastroenterological Surgery, Transplant, and Surgical Oncology, Okayama University, Okayama, Japan.
Naomoto
Yoshio
Y
Nobuhisa
Tetsuji
T
Takaoka
Munenori
M
Motoki
Takayuki
T
Shirakawa
Yasuhiro
Y
Yamatsuji
Tomoki
T
Inoue
Hiroyasu
H
Ouchida
Mamoru
M
Gunduz
Mehmet
M
Nakajima
Motowo
M
Tanaka
Noriaki
N
D000328:Adult; D000368:Aged; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism*; D004938:Esophageal Neoplasms / Q000098:blood supply* / Q000201:enzymology / Q000473:pathology; D004947:Esophagus / Q000201:enzymology / Q000473:pathology; D005260:Female; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009389:Neovascularization, Pathologic; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016019:Survival Analysis; D014162:Transfection; D015854:Up-Regulation / Q000235:genetics; D000099175:Heparanase
D051546:Cyclooxygenase 2; D005966:Glucuronidase; D008156:Luciferases; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D000099175:Heparanase