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Volume 115, Issue 5, Pages (November 1998)

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1 Volume 115, Issue 5, Pages 1090-1095 (November 1998)
A novel gain-of-function mutation of c-kit gene in gastrointestinal stromal tumors  Masanori Nakahara, Koji Isozaki, Seiichi Hirota, Jun-Ichiro Miyagawa, Naoko Hase-Sawada, Masahiko Taniguchi, Toshirou Nishida, Suji Kanayama, Yukihiko Kitamura, Yasuhisa Shinomura, Yuji Matsuzawa  Gastroenterology  Volume 115, Issue 5, Pages (November 1998) DOI: /S (98) Copyright © 1998 American Gastroenterological Association Terms and Conditions

2 Fig. 1 Immunohistochemical characteristics of the tumor. Stained with (A) H&E, (B) anti-KIT antibody, (C) anti-CD34 antibody, (D) antidesmin antibody, (E) anti–α-smooth muscle actin antibody, (F) anti-S100 antibody, and (G) antivimentin antibody. The tumor was positive for KIT, CD34, and vimentin and negative for desmin, α–smooth muscle actin, and S-100 (original magnification 200×). Gastroenterology  , DOI: ( /S (98) ) Copyright © 1998 American Gastroenterological Association Terms and Conditions

3 Fig. 2 Site of the c-kit mutation found in the GIST. Although all mutations observed in 5 GISTs in our previous report were confined within the 11 amino acids (Lys-550 to Val-560) in the juxtamembrane domain, the present GIST showed the deletion-type mutation at codon 579 (Asp) in the juxtamembrane domain. Gastroenterology  , DOI: ( /S (98) ) Copyright © 1998 American Gastroenterological Association Terms and Conditions

4 Fig. 3 Constitutive activation of KITdel579. c-kitdel579 was transfected into Ba/F3 cells. Mouse wild-type c-kit cDNA and c-kitdel559–560 were transfected as controls. (A) Tyrosine phosphorylation was examined with or without rmSCF stimulation. (B) Immune complex kinase assay was performed with or without rmSCF stimulation. KITdel579 showed constitutive phosphorylation and strong kinase activity without stimulation of SCF. Gastroenterology  , DOI: ( /S (98) ) Copyright © 1998 American Gastroenterological Association Terms and Conditions

5 Fig. 4 Autonomous proliferation of Ba/F3 cells with KITdel579 in culture and nude mice. (A) MTT colorimetric assay in the presence or absence of rmIL-3. (B) MTT colorimetric assay in the presence or absence of rmSCF. Data are expressed as the mean of four wells. (C) Development of tumors in nude mice after injection of various Ba/F3 cells. Ba/F3 cells with KITdel579 showed factor-independent growth and formed tumors as in the case of Ba/F3 cells with KITdel559–560. Untransfected Ba/F3 cells (2), Ba/F3 cells with the wild KIT (○), Ba/F3 cells with KITdel559–560 (■), and Ba/F3 cells with KITdel579 (•). For some points, the standard error was too small to be shown by bars. Gastroenterology  , DOI: ( /S (98) ) Copyright © 1998 American Gastroenterological Association Terms and Conditions


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