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Involvement of adaptor protein Crk in malignant feature of human ovarian cancer cell line MCAS

Oncogene volume 25, pages 3547–3556 (2006)Cite this article

Abstract

Signaling adaptor protein Crk regulates cell motility and growth through its targets Dock180 and C3G, those are the guanine-nucleotide exchange factors (GEFs) for small GTPases Rac and Rap, respectively. Recently, overexpression of Crk has been reported in various human cancers. To define the role for Crk in human cancer cells, Crk expression was targeted in the human ovarian cancer cell line MCAS through RNA interference, resulting in the establishment of three Crk knockdown cell lines. These cell lines exhibited disorganized actin fibers, reduced number of focal adhesions, and abolishment of lamellipodia formation. Decreased Rac activity was demonstrated by pull-down assay and FRET-based time-lapse microscopy, in association with suppression of both motility and invasion by phagokinetic track assay and transwell assay in these cells. Furthermore, Crk knockdown cells exhibited slow growth rates in culture and suppressed anchorage-dependent growth in soft agar. Tumor forming potential in nude mice was attenuated, and intraperitoneal dissemination was not observed when Crk knockdown cells were injected into the peritoneal cavity. These results suggest that the Crk is a key component of focal adhesion and involved in cell growth, invasion, and dissemination of human ovarian cancer cell line MCAS.

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Acknowledgements

We thank Dr Michiyuki Matsuda (Osaka University, Japan) for setting up a FRET-based time-lapse microscopy and the providing plasmids for monitoring the activity of small GTPases in a single cell. This study was supported in part by the Japan–China Sasakawa Medical Fellowship, by YASUDA Medical Research Foundation, by Grants-in-Aid from the Ministry of Education, Science, Culture, and Sports, and of the Ministry of Health, Labor, and Welfare.

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Author notes

  1. H Linghu

    Present address: Department of Obstetrics & Gynecology, First Affiliated Hospital of Chongqing University of Medical Sciences, Chongqing, 400012, China

  2. H Sawa

    Present address: Department of Molecular Pathobiology, and 21st Century COE Program for Zoonosis Control, Hokkaido University Research Center for Zoonosis Control, N15, W7, Sapporo, 060-8638, Japan

Authors and Affiliations

  1. Laboratory of Molecular and Cellular Pathology, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, Japan

    H Linghu, M Tsuda, Y Makino, M Sakai, T Watanabe, S Ichihara, H Sawa, K Nagashima & S Tanaka

  2. CREST, Japan Science and Technology Cooporation, Saitama, Japan

    M Tsuda, Y Makino, M Sakai, T Watanabe, S Ichihara, H Sawa, K Nagashima & S Tanaka

  3. National Cardiovascular Center Research Institute, Osaka, Japan

    N Mochizuki

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  1. H Linghu
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Correspondence to S Tanaka.

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Linghu, H., Tsuda, M., Makino, Y. et al. Involvement of adaptor protein Crk in malignant feature of human ovarian cancer cell line MCAS. Oncogene 25, 3547–3556 (2006). https://doi.org/10.1038/sj.onc.1209398

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