title: The ubiquitin E3 ligase WWP1 decreases CXCL12-mediated MDA231 breast cancer cell migration and bone metastasis.
abstract: Advanced breast cancers preferentially metastasize to bone where cells in the bone microenvironment produce factors that enhance breast cancer cell homing and growth. Expression of the ubiquitin E3 ligase WWP1 is increased in some breast cancers, but its role in bone metastasis has not been investigated. Here, we studied the effects of WWP1 and itch, its closest family member, on breast cancer bone metastasis. First, we immunostained a multi-tumor tissue microarray and a breast cancer tissue microarray and demonstrated that WWP1 and ITCH are expressed in some of breast cancer cases. We then knocked down WWP1 or itch in MDA-MB-231 breast cancer cells using shRNA and inoculated these cells and control cells into the left ventricle of athymic nude mice. Radiographs showed that mice given shWWP1 cells had more osteolytic lesions than mice given control MDA-MB-231 cells. Histologic analysis confirmed osteolysis and showed significantly increased tumor area in bone marrow of the mice. WWP1 knockdown did not affect cell growth, survival or osteoclastogenic potential, but markedly increased cell migration toward a CXCL12 gradient in vitro. Furthermore, WWP1 knockdown significantly reduced CXCL12-induced CXCR4 lysosomal trafficking and degradation. In contrast, itch knockdown had no effect on MDA-MB-231 cell bone metastasis. Taken together, these findings demonstrate that WWP1 negatively regulates cell migration to CXCL12 by limiting CXCR4 degradation to promote breast cancer metastasis to bone and highlight the potential utility of WWP1 as a prognostic indicator for breast cancer bone metastasis.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, NY 14642, USA.
authors.forename: Kristina
authors.initials: K
authors.lastname: Subik
authors.forename: Lei
authors.initials: L
authors.lastname: Shu
authors.forename: Chengyu
authors.initials: C
authors.lastname: Wu
authors.forename: Qianqian
authors.initials: Q
authors.lastname: Liang
authors.forename: David
authors.initials: D
authors.lastname: Hicks
authors.forename: Brendan
authors.initials: B
authors.lastname: Boyce
authors.forename: Linda
authors.initials: L
authors.lastname: Schiffhauer
authors.forename: Di
authors.initials: D
authors.lastname: Chen
authors.forename: Ceshi
authors.initials: C
authors.lastname: Chen
authors.forename: Ping
authors.initials: P
authors.lastname: Tang
authors.forename: Lianping
authors.initials: L
authors.lastname: Xing
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001859:Bone Neoplasms / Q000150:complications / Q000473:pathology / Q000556:secondary*; D001943:Breast Neoplasms / Q000150:complications / Q000201:enzymology* / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D054377:Chemokine CXCL12 / Q000494:pharmacology*; D005260:Female; D055785:Gene Knockdown Techniques; D006801:Humans; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D010014:Osteolysis / Q000150:complications / Q000473:pathology; D021381:Protein Transport / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000378:metabolism; D011960:Receptors, Estrogen / Q000378:metabolism; D011980:Receptors, Progesterone / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D046888:Tissue Array Analysis; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
chemical_list: C527276:CXCR4 protein, human; D054377:Chemokine CXCL12; D034741:RNA, Small Interfering; D019718:Receptors, CXCR4; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D012097:Repressor Proteins; C432966:ITCH protein, human; C471642:WWP1 protein, human; D044767:Ubiquitin-Protein Ligases