title: Cbl-b accelerates trypsin-induced cell detachment through ubiquitination and degradation of proline-rich tyrosine kinase 2.
abstract: Trypsin is a digestive enzyme that is widely used for cell detachment, which is the first stage of tumor metastasis. Recent studies show that adhesion-related kinases are involved in cell detachment. Proline-rich tyrosine kinase 2 (Pyk2) is a crucial kinase in the regulation of cell adhesion and detachment. However, the effect of Pyk2 on cell detachment is controversial. In the present study, we found that Pyk2 expression was rapidly decreased after trypsin treatment in gastric cancer, breast cancer, colon cancer, lung cancer, and human gastric epithelial cells. Knockdown of Pyk2 accelerated cell detachment. Furthermore, lysosome inhibitor NH4CL suppressed cell detachment and increased ubiquitination of Pyk2. Cbl-b is a type of E3 ubiquitin ligase that interacted with Pyk2, reduced the expression of Pyk2, and promoted trypsin-induced degradation of Pyk2. These findings suggest that Cbl-b promoted cell detachment through mono-ubiquitination of Pyk2. Our data provide a new insight into the role of Cbl-b in cell detachment.
authors.affiliation: Department of Medical Oncology, The First Hospital of China Medical University, No.155, North Nanjing Street, Heping District, Shenyang, 110001, China.
authors.forename: Yibo
authors.initials: Y
authors.lastname: Fan
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Qu
authors.forename: Yanju
authors.initials: Y
authors.lastname: Ma
authors.forename: Jinglei
authors.initials: J
authors.lastname: Qu
authors.forename: Yunpeng
authors.initials: Y
authors.lastname: Liu
authors.forename: Xuejun
authors.initials: X
authors.lastname: Hu
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D051418:Focal Adhesion Kinase 2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D009369:Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D010766:Phosphorylation / Q000187:drug effects; D059748:Proteolysis / Q000187:drug effects*; D050721:Proto-Oncogene Proteins c-cbl / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D014357:Trypsin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D054875:Ubiquitination / Q000187:drug effects*
chemical_list: D034741:RNA, Small Interfering; D050721:Proto-Oncogene Proteins c-cbl; D051418:Focal Adhesion Kinase 2; D014357:Trypsin
references.citation: Eur J Cancer. 2012 Nov;48(17):3288-99
references.pmid: 22456178
references.citation: FEBS Lett. 2009 Jul 7;583(13):2255-62
references.pmid: 19508871
references.citation: J Immunol. 2010 Jul 15;185(2):917-28
references.pmid: 20543098
references.citation: Bone. 2007 May;40(5):1329-42
references.pmid: 17321230
references.citation: Br J Cancer. 2009 May 19;100(10):1540-8
references.pmid: 19384300
references.citation: Oncogene. 2004 Dec 2;23(56):9052-61
references.pmid: 15489898
references.citation: J Pathol. 2006 Jun;209(2):147-56
references.pmid: 16691544
references.citation: J Colloid Interface Sci. 2012 Sep 1;381(1):125-36
references.pmid: 22727404
references.citation: J Cell Biochem. 2009 Nov 1;108(4):851-9
references.pmid: 19711372
references.citation: Biochem Biophys Res Commun. 2008 Feb 29;367(1):183-9
references.pmid: 18164258
references.citation: J Biol Chem. 2012 Feb 17;287(8):5327-39
references.pmid: 22203672
references.citation: Biochem Biophys Res Commun. 2012 Feb 3;418(1):144-8
references.pmid: 22244876
references.citation: Leuk Res. 2013 Aug;37(8):956-62
references.pmid: 23587524
references.citation: FEBS Lett. 2012 Sep 21;586(19):3104-10
references.pmid: 22884570
references.citation: Cell Signal. 2007 Feb;19(2):289-300
references.pmid: 16945503
references.citation: J Biol Chem. 2007 Aug 17;282(33):24120-30
references.pmid: 17581824
references.citation: J Cell Biol. 2000 Mar 20;148(6):1267-81
references.pmid: 10725339
references.citation: PLoS One. 2012;7(6):e40087
references.pmid: 22768227
references.citation: J Cell Sci. 2004 May 15;117(Pt 12):2557-68
references.pmid: 15128873
references.citation: BMC Cancer. 2010 Sep 17;10:498
references.pmid: 20849596