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J. Biol. Chem., Vol. 281, Issue 50, 38941-38950, December 15, 2006

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Expression of Claudin7 Is Tightly Associated with Epithelial Structures in Synovial Sarcomas and Regulated by an Ets Family Transcription Factor, ELF3^*

Yoshiki Kohno, Takeshi Okamoto, Tatsuya Ishibe, Satoshi Nagayama^¶, Yasuko Shima, Kohichi Nishijo, Kotaro R. Shibata, Kenichi Fukiage, Seiji Otsuka^||, Daisuke Uejima^**, Nobuhito Araki, Norifumi Naka, Yasuaki Nakashima, Tomoki Aoyama, Tomitaka Nakayama, Takashi Nakamura, and Junya Toguchida¹

From the Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, the Department of Orthopaedic Surgery and the ^¶Department of Surgical Oncology, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, the ^||Department of Musculoskeletal Medicine, Graduate School of Medical Sciences, Nagoya City University, Aichi 467-8601, the ^**Department of Orthopedic Surgery, Kansai Medical University, Osaka 570-8506, the Department of Orthopedic Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, Osaka 537-8511, and the Department Pathology, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan

Synovial sarcoma, a soft tissue sarcoma that develops in adults, is pathologically subclassified into monophasic spindle synovial sarcoma and biphasic synovial sarcoma with epithelial components. The molecular mechanism building the epithelial components in biphasic synovial sarcoma is totally unknown. Here we investigated claudins, critical molecules in the tight junction, in biphasic synovial sarcoma. Expression profiles of 21 claudins in 17 synovial sarcoma tumor samples, including 9 biphasic tumors, identified claudin4, claudin7, and claudin10 as biphasic tumor-related claudins, and immunohistochemical analyses demonstrated the localization of these claudins in the epithelial component in biphasic tumors, with claudin7 the most closely associated with the epithelial component. The mRNA expression and protein localization of claudin7 coincided with those of the ELF3, an epithelia-specific member of the Ets family of transcription factors. Luciferase reporter assays demonstrated that the presence of the Ets-binding site at -150 in the promoter region of the claudin7 gene was critical for the transcriptional activity, and gel shift and chromatin immunoprecipitation assays confirmed the binding of ELF3 to the Ets site at -150. Inhibition of ELF3 expression by small interfering RNA simultaneously down-regulated the mRNA expression of the claudin7 gene, and the introduction of ELF3 expression in claudin7-negative cell lines induced mRNA expression of the claudin7 gene. Therefore, the induction of claudin7 expression by ELF3 appears critical to the formation of the epithelial structures in biphasic synovial sarcoma.

Received for publication, August 31, 2006 , and in revised form, October 18, 2006.

^* This work was supported by grants-in-aid for scientific research from the Japan Society for the Promotion of Science, from the Ministry of Education, Culture, Sports, Science, and Technology, and from the Ministry of Health, Labor, and Welfare. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. Tel.: 81-75-751-4134; Fax: 81-75-751-4144; E-mail: togjun{at}frontier.kyoto-u.ac.jp.

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