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Effects of 4-(3-Chloro-Benzyl)-6,7-Dimethoxy-Quinazoline on Kinetics of P120-Catenin and Periplakin in Human Buccal Mucosa Squamous Carcinoma Cell Line

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Open Journal of Stomatology 2014年第5期4卷 249-257页

作者： Isao Tamura Aiko Kamada Seiji Goda Yoshihiro Yoshikawa Eisuke Domae takashi ikeo Department of Biochemistry Osaka Dental University Hirakata Japan

In order to detect molecular markers for the epidermal growth factor inhibitor 4-(3-chloro-benzyl)- 6,7-dimethoxy-quinazoline (tyrphostin), we investigated the kinetics of p120-catenin and periplakin in the human buccal mucosa squamous cancer cell line BICR 10 treated with 3 nM tyrphostin. Growth of BICR 10 cells was inhibited by treatment with tyrphostin. Although changes were not observed in the expression of EGFR and p120-catenin, expression of Akt, Src and periplakin in BICR 10 treated with 3 nM tyrphostin tended to decrease. In addition, phosphorylation of EGFR, Akt and Src was inhibited by treatment with tyrphostin. On immunocytochemical staining, immunoreactions with phosphorylated EGFR, phosphorylated Akt and phosphorylated p120-catenin were weak in BICR 10 treated with tyrphostin. There was a slight immunocy to chemical reaction to periplakin in BICR 10 cells induced by tyrphostin. In conclusion, the decrease in phosphorylation in EGFR and p120-catenin by tyrphostin, following the decrease in Src or Akt phosphorylation, may inhibit expression of several growth factors associated with the proliferation and migration of cancer cells.

关键词： 4-(3-Chloro-Benzyl)-6 7-Dimethoxy-Quinazoline Human Buccal Mucosa Squamous Cancer Cell Line P120-Catenin Periplakin

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Influence of Adiponectin and Leptin on the Kinetics of Human Pulp Cells

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Open Journal of Stomatology 2020年第12期10卷 360-371页

作者： Isao Tamura Katsura Ueda Yoshifumi Matsuda Aiko Kamada Yoshihiro Yoshikawa Eisuke Domae takashi ikeo Shunji Kumabe Department of Oral Anatomy School of Dentistry Osaka Dental University 8-1 Kuzuhahanazono-cho Hirakata-shi Osaka Japan Department of Biochemistry School of Dentistry Osaka Dental University 8-1 Kuzuhahanazono-cho Hirakata-shi Osaka Japan

Adipocytokines, such as adiponectin and leptin, are expressed by adipocytes and are known as anti-metabolic syndrome factors. They are also thought to mediate calcification in mineralized tissue. We investigated the effects of adiponectin and leptin on the kinetics of human pulp cells using ELISA and western blot. After gaining informed consent, we obtained human pulp cells from three patients. After cultivation in Dulbecco’s Modified Eagle Medium (DMEM) without serum, cells from the 4th to 6th passages were incubated with various concentrations of adiponectin (1, 10, or 100 ng/mL) in DMEM or leptin (0.1, 1, or 10 ng/mL) in DMEM for 24 h. We confirmed that human pulp cells expressed adiponectin receptor 1 and leptin receptor. Although the proliferation index of these cells as measured by 5-bromo-2'-deoxyuridine (BrdU) incorporation increased in the presence of adiponectin in a dose-dependent manner, pulp cells stimulated with leptin showed no significant changes in BrdU incorporation. Alkaline phosphatase activity showed no significant changes after stimulation with either adipocytokine. Adiponectin induced expression of BMP-2 and osteopontin more strongly than did leptin. In particular, expression of BMP-2 increased in the presence of adiponectin in a dose-dependent manner. In contrast, the expression of dentin sialoprotein increased after stimulation with leptin. The expression of Runx2 was not observed from the cultured pulp cells stimulated with both molecules. These results indicate that adiponectin and leptin contribute to growth and differentiation of human pulp cells and may consequently affect the formation of secondary dentin or reparative dentin in the dentin-pulp complex.

关键词： Dental Pulp Cells Adiponectin Leptin Bone Morphogenetic Protein-2 Dentin Sialoprotein Osteopontin ECM Formation

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Expression of Plectin-1 and Trichohyalin in Human Tongue Cancer Cells

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Open Journal of Stomatology 2018年第6期8卷 196-204页

作者： Isao Tamura Katsura Ueda Tetsunari Nishikawa Aiko Kamada Tomoharu Okamura Yoshifumi Matsuda Kentaro Ueno Yoshihiro Yoshikawa Eisuke Domae Kazuya Tominaga Shunji Kumabe takashi ikeo Akio Tanaka Department of Oral Anatomy School of Dentistry Osaka Dental University Hirakata Japan Department of Innovation in Dental Education School of Dentistry Osaka Dental University Hirakata Japan Department of Biochemistry School of Dentistry Osaka Dental University Hirakata Japan Department of Oral Pathology School of Dentistry Osaka Dental University Hirakata Japan Department of Pathology School of Dentistry Osaka Dental University Hirakata Japan

In basal squamous cells, plectin-1 interacts with intermediate filaments, whereas trichohyalin, which is distributed primarily in the medulla and inner root sheath cells of human hair follicles, plays a role in strengthening cells during keratinization. Although both cytoskeletal proteins occur in trace amounts in human tongue epithelial cells, there are minimal data on their expression in human tongue primary cancer cells. We therefore investigated the expression of plectin-1 and trichohyalin in human tongue epithelial cell line (DOK) and tongue cancer cell line (BICR31) using western blotting and FITC-labeled immunocytochemistry techniques. DOK and BICR31 cells were cultivated to subconfluence in Dulbecco’s Modified Eagle’s Medium containing 0.4 μg/ml of hydrocortisone and 10% fetal bovine serum, and the levels of trichohyalin and plectin-1 were determined by western blot analysis and immunocytochemical staining. Trichohyalin expression was clearly observed, with no differences between DOK and BICR31 cells. Although DOK cells expressed trace levels of plectin-1, obvious plectin-1 bands were detected in western blot analyses of BICR31 cells. Immunocytochemical staining revealed that trichohyalin and plectin-1 localize in the cytoplasm. Trichohyalin was diffusely distributed in both cell lines, and colocalization of trichohyalin and cytokeratin 1/10 was observed in almost all BICR31 cells. There were no correlations between western blot and immunocytochemical data for trichohyalin. Conversely, correlations in immunochemical reactions for plectin-1 were observed. Most DOK cells showed no localization of plectin-1, but strong reactions were detected in the cytoplasm of BICR31 cells. These results indicate that trichohyalin is expressed by cancerous tongue epithelial cells during various stages of malignancy and that plectin-1 provides an index of malignancy.

关键词： Tongue Cancer Plectin-1 Trichohyalin Diagnosis

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