Abstract 847: Anti-cancer effects of selective Cox-2 and EP2 inhibition through suppression of EMT and the clinical implications of overexpression of Cox-2 and downregulation of E-cadherin in pharyngeal squamous cell carcinoma
Authors: Watanabe Yoshihiro , Imanishi Yorihisa , Ozawa Hiroyuki , Kameyama Kaori , Sakamoto Koji , Fujii Ryoichi , Shigetomi Seiji , Habu Noboru , Otsuka Kuninori , Sato Yoichiro , Sekimizu Mariko , Ito Fumihiro , Ikari Yuichi , Saito Shin , Ogawa Kaoru 
Published: July 2017

Background: Overexpression of cyclooxygenase 2 (Cox-2), an inducible prostaglandin (PG) synthetase, is assumed to promote cancer progression through its multifaceted function, including induction of angiogenesis, stimulation of cell proliferation, restraint on apoptosis, and immunosuppression. Furthermore, recently its inverse relationship with E-cadherin expression was reported. The epithelial-to-mesenchymal transition (EMT) accompanied by the downregulation of E-cadherin is supposed to promote metastasis. However, neither the anti-cancer effect of selective Cox-2 inhibitors on the regulation of E-cadherin nor its specific mechanism has been examined in pharyngeal squamous cell carcinoma (PhSCC). In addition, the anti-tumor effect of EP2 antagonist, a possibly more selective inhibitor of PGE2, remains to be elucidated.

Methods: We used quantitative real-time PCR to examine the effects of a selective Cox-2 inhibitors, Celecoxib, and a selective EP2 antagonist, PF-04418948, on the gene expressions of CDH-1, its transcriptional repressors (snail, ZEB1, twist), and vimentin in the human PhSCC cell lines BICR6 and FaDu. Alteration in E-cadherin expression on the cell surface was evaluated by immunofluorescent staining. In vitro proliferation and migration assays were performed to assess cellular behaviors. Clinicopathological factors and immunohistochemical expressions of Cox-2 and E-cadherin were evaluated using surgical specimens from 54 patients with PhSCC who underwent transoral resection.

Results: Both Celecoxib and PF-04418948 upregulated the CDH-1 expression and the membranous E-cadherin expression and downregulate the vimentin expression in the PhSCC cells through the suppression of snail, ZEB1, and twist expression. The extent of this effect depended on the baseline expression levels of both E-cadherin and Cox-2 in each cell line. Additionally, the cell migration was attenuated by both Celecoxib and PF-04418948, whereas the cell proliferation was not affected. Univariate analysis showed that differentiation, overexpression of Cox-2, and decreased expression of E-cadherin were significantly correlated with lymph node metastasis. Multivariate logistic regression revealed that overexpression of Cox-2 (odds ratio [OR]=53.49, P&lt;0.001) and decreased expression of E-cadherin (OR=0.06, P=0.016) were the independent risk factors affecting lymph node metastasis.

Conclusions: These findings suggest that the appropriately selective administration of the Cox-2 inhibitor and EP2 antagonist may have an anti-metastatic effect through suppression of EMT by restoring E-cadherin expression. In addition, overexpression of Cox-2 and downregulation of E-cadherin may be closely implicated in lymph node metastasis in PhSCC.

Citation Format: Yoshihiro Watanabe, Yorihisa Imanishi, Hiroyuki Ozawa, Kaori Kameyama, Koji Sakamoto, Ryoichi Fujii, Seiji Shigetomi, Noboru Habu, Kuninori Otsuka, Yoichiro Sato, Mariko Sekimizu, Fumihiro Ito, Yuichi Ikari, Shin Saito, Kaoru Ogawa. Anti-cancer effects of selective Cox-2 and EP2 inhibition through suppression of EMT and the clinical implications of overexpression of Cox-2 and downregulation of E-cadherin in pharyngeal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 847. doi:10.1158/1538-7445.AM2017-847