Noboru Habu

Restoration of E-cadherin expression by selective Cox-2 inhibition and the clinical relevance of the epithelial-to-mesenchymal transition in head and neck squamous cell carcinoma

BackgroundThe epithelial-to-mesenchymal transition (EMT) accompanied by the downregulation of E-cadherin has been thought to promote metastasis. Cyclooxygenase-2 (Cox-2) is presumed to contribute to cancer progression through its multifaceted function, and recently its inverse relationship with E-cadherin was suggested. The aim of the present study was to investigate whether selective Cox-2 inhibitors restore the expression of E-cadherin in head and neck squamous cell carcinoma (HNSCC) cells, and to examine the possible correlations of the expression levels of EMT-related molecules with clinicopathological factors in HNSCC.MethodsWe used quantitative real-time PCR to examine the effects of three selective Cox-2 inhibitors, i.e., celecoxib, NS-398, and SC-791 on the gene expressions of E-cadherin (CDH-1) and its transcriptional repressors (SIP1, Snail, Twist) in the human HNSCC cell lines HSC-2 and HSC-4. To evaluate the changes in E-cadherin expression on the cell surface, we used a flowcytometer and immunofluorescent staining in addition to Western blotting. We evaluated and statistically analyzed the clinicopathological factors and mRNA expressions of Cox-2, CDH-1 and its repressors in surgical specimens of 40 patients with tongue squamous cell carcinoma (TSCC).ResultsThe selective Cox-2 inhibitors upregulated the E-cadherin expression on the cell surface of the HNSCC cells through the downregulation of its transcriptional repressors. The extent of this effect depended on the baseline expression levels of both E-cadherin and Cox-2 in each cell line. A univariate analysis showed that higher Cox-2 mRNA expression (p = 0.037), lower CDH-1 mRNA expression (p = 0.020), and advanced T-classification (p = 0.036) were significantly correlated with lymph node metastasis in TSCC. A multivariate logistic regression revealed that lower CDH-1 mRNA expression was the independent risk factor affecting lymph node metastasis (p = 0.041).ConclusionsThese findings suggest that the appropriately selective administration of certain Cox-2 inhibitors may have an anti-metastatic effect through suppression of the EMT by restoring E-cadherin expression. In addition, the downregulation of CDH-1 resulting from the EMT may be closely involved in lymph node metastasis in TSCC.

QOL after head and neck reconstruction: Evaluation of Japanese patients using SF-36 and GOHAI

OBJECTIVE To evaluate the health-related quality of life (HRQOL) of Japanese patients with head and neck cancer after treatment. STUDY DESIGN Cross-sectional, descriptive study. PATIENTS AND METHODS Twenty-nine patients who underwent resection and reconstruction of the head and neck cancer between September 2001 and January 2008 at the National Hospital Organization Tokyo Medical Center completed the Short Form 36 (generic QOL measure) and the General Oral Health Assessment Index (oral-specific QOL measure). RESULTS The generic QOL of the patients was relatively maintained while oral-specific QOL was impaired compared to the Japanese norms. The patients with musculo-cutaneous flaps and 1y or longer after operation reported significantly lower QOL. CONCLUSION This is a unique study on Japanese patients with relatively longer time after operation. Further evaluation with increased number of cases and disease-specific QOL scale is required to better understand the QOL of the patients.

Clinical diagnosis and treatment outcomes for parapharyngeal space schwannomas: A single-institution review of 21 cases

Because the incidence of schwannoma arising from the parapharyngeal space (PPS) is very low, no studies have analyzed extirpation methods and postoperative neurological complications exclusively in PPS schwannomas.

Abstract 2818: Flt-4 expression on tumor cells and its autocrine mechanism with VEGF-C/contactin-1 stimulate progression of head and neck squamous cell carcinoma.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Background: Tumor cell-derived VEGF-C has been considered to promote lymphangiogenesis by activating Flt-4 (VEGFR-3) expressed on lymphatic endothelial cells via a paracrine mechanism in tumor microenvironment. Recent studies have shown that Flt-4 is expressed not only in endothelial cells but also selectively in certain subsets of a variety of cancer cells. In addition, the VEGF-C/Flt-4 autocrine loop on lung adenocarcinoma cells was reported to enhance cell motility and invasiveness through up-regulation of the neural cell adhesion molecule Contactin-1 (CNTN-1), although little of such mechanism is known in head and neck squamous cell carcinoma (HNSCC). Present study was aimed to examine Flt-4 expression and VEGF-C/Flt-4 autocrine loop in HNSCC cells, and clinicopathological significance of the expressions of Flt-4, VEGF-C, and CNTN-1 in the tumor specimens of the patients with HNSCC. Methods: We examined expression of Flt-4 in HNSCC cell lines using quantitative real-time PCR, RT-PCR, and immunocytochemistry. The role of the VEGF-C/Flt-4 axis in Flt-4-positive HNSCC cells in vitro was investigated by stimulating or blocking Flt-4. Immunohistochemical studies were performed on 56 oral tongue squamous cell carcinoma (TSCC) specimens. The associations of the immunohistochemical expressions of Flt-4, VEGF-C, and CNTN-1 with the clinicopathological factors were statistically analyzed. Results: Real-time PCR detected Flt-4 mRNA expression in several HNSCC cells, which was further confirmed by RT-PCR and immunocytochemistry in SAS and HO1U1. Stimulation of Flt-4 using recombinant VEGF-C up-regulated expression of CNTN-1 and VEGF-C itself in SAS, whereas blocking using neutralizing anti-Flt-4 antibody or Flt-4 inhibitor down-regulated expression of CNTN-1 in both SAS and HO1U1, and that of VEGF-C itself in SAS. Immunohistochemical analyses using TSCC specimens found significant correlation of CNTN-1 expression with both VEGF-C and Flt-4 expressions, but not between VEGF-C and Flt-4. With regard to clinicopathological factors, expressions of all the three molecular markers in tumor cells significantly correlated with lymph node (LN) metastasis, which was also correlated with T classification and lymphatic invasion in univariate analysis. Multivariate analysis elucidated that T classification (p=0.003), lymphatic invasion (p=0.024), and expression of Flt-4 in tumor cells (p=0.046) were independent predictors of LN metastasis. Conclusions: Our results suggest that VEGF-C activates FLt-4 expressed not only on lymphatic endothelial cells but also selectively on subset of HNSCC cells, resulting in VEGF-C/Flt-4 autocrine mechanism that up-regulates CNTN-1 and VEGF-C itself in tumor cells. Therefore, both paracrine and autocrine mechanisms of VEGF-C/Flt-4 may contribute to progression of HNSCC including LN metastasis. Citation Format: Seiji Shigetomi, Yorihisa Imanishi, Masayuki Shimoda, Kaori Kameyama, Katsushi Shibata, Nobuya Sakai, Koji Sakamoto, Ryoichi Fujii, Noboru Habu, Kuninori Ootsuka, Kaoru Ogawa. Flt-4 expression on tumor cells and its autocrine mechanism with VEGF-C/contactin-1 stimulate progression of head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2818. doi:10.1158/1538-7445.AM2013-2818

Abstract 3359: Expression of stem cell markers Oct3/4 and Nanog in the head and neck squamous carcinoma cells and its clinical implications for delayed neck metastasis in stage I/II tongue squamous cell carcinoma

Background: The side population (SP) of cancer cells is reportedly enriched with cancer stem cells (CSCs), however, the functional role and clinical relevance of CSC marker molecules upregulated in the SP of head and neck squamous carcinoma (HNSCC) cells are yet to be elucidated. Patients with clinical stage I/II (T1-2N0M0) tongue squamous cell carcinoma (TSCC) typically undergo partial glossectomy alone; however, development of delayed neck metastasis (DNM) tends to reduce their survival. In the present study, we aimed to determine the CSC markers in the SP of HNSCC cells along with their functions in cellular behaviors, and to clarify the association of these markers with DNM. Methods: Flow cytometry was applied to isolate SP from main population (MP) in HNSCC cells (SCC4, SAS, and HSC4). The expression of the CSC markers was examined by semi-quantitative RT-PCR and immunocytochemistry. In vitro proliferation, migration, and invasion assays were performed to assess cellular behaviors. Clinicopathological factors and immunohistochemical expressions of Oct3/4 and Nanog were evaluated using surgical specimens from 50 patients with stage I/II TSCC who underwent partial glossectomy alone. Results: SPs were isolated in all three cell lines examined. Expression levels of Oct3/4 and Nanog were higher in SP cells than MP cells. Additionally, cell migration and invasion abilities were higher in SP cells than MP cells, whereas there was no difference in proliferation. Univariate analysis showed that expression of Oct3/4 and Nanog, vascular invasion, muscular invasion, and mode of invasion were significantly correlated with DNM. Multivariate logistic regression revealed that Oct3/4 expression (risk ratio = 14.78, p = 0.002) and vascular invasion (risk ratio = 12.93, p = 0.017) were independently predictive of DNM. Regarding the diagnostic performance, Oct3/4 showed the highest accuracy, sensitivity, and negative predictive value of 82.0%, 61.5%, and 86.8%, respectively, while vascular invasion showed the highest specificity and positive predictive value of 94.6% and 71.4%, respectively. Conclusion: These results suggest that Oct3/4 and Nanog represent probable CSC markers in HNSCC, which contribute to the development of DNM in part by enhancing cell motility and invasiveness. Moreover, along with vascular invasion, expression of Oct3/4 in the primary cancer cells can be considered a potential predictor for selecting patients at high risk of developing DNM. Citation Format: Noboru Habu, Yorihisa Imanishi, Kaori Kameyama, Masayuki Shimoda, Yutaka Tokumaru, Koji Sakamoto, Ryoichi Fujii, Seiji Shigetomi, Kuninori Otsuka, Yoichiro Sato, Yoshihiro Watanabe, Hiroyuki Ozawa, Toshiki Tomita, Masato Fujii, Kaoru Ogawa. Expression of stem cell markers Oct3/4 and Nanog in the head and neck squamous carcinoma cells and its clinical implications for delayed neck metastasis in stage I/II tongue squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3359.