Tumor cell migration is a key step underlying cancer cell dissemination and metastasis and is controlled by extracellular signaling-mediated dynamic cytoskeletal and cell matrix adhesion remodeling. Using a phagokinetic track (PKT) assay in combination with multi-parametric image analysis and highly motile H1299 adenocarcinoma cells, we screened for 1,429 upstream kinase signaling components and downstream adhesion and cytoskeletal regulators that determine tumor cell migratory behavior: speed, directionality, and persistence. Thirty significant genes were validated by live cell imaging random tumor cell migration, which was associated with modulation of focal adhesion dynamics. For eight genes a significant association with metastasis free survival in breast cancer patients was observed, SHC1, SRPK1, NEK2, ITGB3BP and MAP3K8 being most significant. Also, high SRFS protein kinase 1 (SRPK1) protein expression on breast cancer tissue micro arrays was associated with poor disease outcome. SRPK1 expression was highest in basal-like breast cancer cell lines and depletion of SRPK1 inhibited breast cancer cell motility and focal adhesion dynamics. Finally, in an orthotopic mammary tumor metastasis model, stable knockdown of SRPK1 in lung metastatic variant MDA-MB-231 basal-like breast cancer cells reduced lung metastasis formation. This study provides a comprehensive information resource on the molecular determinants of tumor cell migration in close association with a clinical significant role in breast cancer progression.
Publication:
A Kinome Screen Identifies SRPK1 to Mediate Breast Cancer Metastasis
Authors:
Wies van Roosmalen, Sylvia E. le Dévédec, Ofra Golani, Marcel Smid, Irina Pulyakhina, Annemieke M. Timmermans, Maxime P. Look, Zi Di, Chantal Pont, Marjo de Graauw, Suha Naffar-Abu-Amara, Peter A.C. 't Hoen, John W.M. Martens, John A. Foekens, Benjamin Geiger, Bob van de Water
