Reza Salimi1,2, Sashidhar Bandaru1, Sravani Devarakonda1, Sevtap Gökalp1, Chandu Ala1, Ali Alvandian2, Nilgün Yener3, Levent M Akyürek4,5. 1. Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden. 2. Department of Molecular Medicine, Faculty of Medicine, Dokuz Eylül University, Izmir, Turkey. 3. Department of Medical Biochemistry, Faculty of Medicine, Dokuz Eylül University, Izmir, Turkey. 4. Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden levent.akyurek@gu.se. 5. Department of Clinical Pathology and Genetics, Sahlgrenska Academy Hospital, Gothenburg, Sweden.
Abstract
BACKGROUND/AIM: Filamin A (FLNA) is the most abundant and widely expressed isoform of filamin in human tissues. It is cleaved by calpain at the hinge 1 and 2 domains, producing a 90-kDa carboxyl-terminal fragment (FLNACT). Recently, it has been shown that FLNACT mediates cell signaling and transports transcription factors into the cell nucleus. However, the significance of cleavage of FLNA by calpain has not been studied in cancer cell growth. Calpeptin is a chemical inhibitor of both calpain 1 and 2 that cleaves FLNA. In this study, we questioned if inhibiting calpain using calpeptin would decrease tumor cell proliferation, migration, invasion, and colony formation. MATERIALS AND METHODS: Human melanoma (A7), prostate cancer (PC3), mouse fibrosarcoma (T241) and endothelial (MS1) cells were assayed for proliferation, migration, invasion and colony formation after treatment with calpeptin. Cell lysates were immunoblotted for FLNA and FLNACT Results: Calpeptin treatment of these cells resulted in a decreased production of FLNACT Calpeptin-treated human and mouse tumor cells displayed impaired proliferation, migration, and colony formation. CONCLUSION: These data suggest that the cleavage of FLNA by calpain is an important cellular event in the regulation of tumor cell growth. Copyright
BACKGROUND/AIM: Filamin A (FLNA) is the most abundant and widely expressed isoform of filamin in human tissues. It is cleaved by calpain at the hinge 1 and 2 domains, producing a 90-kDa carboxyl-terminal fragment (FLNACT). Recently, it has been shown that FLNACT mediates cell signaling and transports transcription factors into the cell nucleus. However, the significance of cleavage of FLNA by calpain has not been studied in cancer cell growth. Calpeptin is a chemical inhibitor of both calpain 1 and 2 that cleaves FLNA. In this study, we questioned if inhibiting calpain using calpeptin would decrease tumor cell proliferation, migration, invasion, and colony formation. MATERIALS AND METHODS:Humanmelanoma (A7), prostate cancer (PC3), mousefibrosarcoma (T241) and endothelial (MS1) cells were assayed for proliferation, migration, invasion and colony formation after treatment with calpeptin. Cell lysates were immunoblotted for FLNA and FLNACT Results:Calpeptin treatment of these cells resulted in a decreased production of FLNACT Calpeptin-treated human and mousetumor cells displayed impaired proliferation, migration, and colony formation. CONCLUSION: These data suggest that the cleavage of FLNA by calpain is an important cellular event in the regulation of tumor cell growth. Copyright