Tumor cell migration screen identifies SRPK1 as breast cancer metastasis determinant
Wies van Roosmalen, … , Benjamin Geiger, Bob van de Water
Wies van Roosmalen, … , Benjamin Geiger, Bob van de Water
Published April 1, 2015; First published March 16, 2015
Citation Information: J Clin Invest. 2015;125(4):1648-1664. https://doi.org/10.1172/JCI74440.
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Categories: Research Article Oncology

Tumor cell migration screen identifies SRPK1 as breast cancer metastasis determinant

Abstract

Tumor cell migration is a key process for cancer cell dissemination and metastasis that is controlled by signal-mediated cytoskeletal and cell matrix adhesion remodeling. Using a phagokinetic track assay with migratory H1299 cells, we performed an siRNA screen of almost 1,500 genes encoding kinases/phosphatases and adhesome- and migration-related proteins to identify genes that affect tumor cell migration speed and persistence. Thirty candidate genes that altered cell migration were validated in live tumor cell migration assays. Eight were associated with metastasis-free survival in breast cancer patients, with integrin β3–binding protein (ITGB3BP), MAP3K8, NIMA-related kinase (NEK2), and SHC-transforming protein 1 (SHC1) being the most predictive. Examination of genes that modulate migration indicated that SRPK1, encoding the splicing factor kinase SRSF protein kinase 1, is relevant to breast cancer outcomes, as it was highly expressed in basal breast cancer. Furthermore, high SRPK1 expression correlated with poor breast cancer disease outcome and preferential metastasis to the lungs and brain. In 2 independent murine models of breast tumor metastasis, stable shRNA-based SRPK1 knockdown suppressed metastasis to distant organs, including lung, liver, and spleen, and inhibited focal adhesion reorganization. Our study provides comprehensive information on the molecular determinants of tumor cell migration and suggests that SRPK1 has potential as a drug target for limiting breast cancer metastasis.

Authors

Wies van Roosmalen, Sylvia E. Le Dévédec, Ofra Golani, Marcel Smid, Irina Pulyakhina, Annemieke M. Timmermans, Maxime P. Look, Di Zi, Chantal Pont, Marjo de Graauw, Suha Naffar-Abu-Amara, Catherine Kirsanova, Gabriella Rustici, Peter A.C. ‘t Hoen, John W.M. Martens, John A. Foekens, Benjamin Geiger, Bob van de Water

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Figure 8

Knockdown of SRPK1 does not affect splicing but results in the differential expression of almost 200 genes when analyzed with deep sequencing.

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Knockdown of SRPK1 does not affect splicing but results in the different...
(A) No difference in splicing of MAP2K2 and VEGFA was detected. (B) Pairwise comparison to identify common genes between the 4 groups that are significantly differentially expressed (log cpm >1.4; FDR <0.1). (C) A pathway analysis performed with MetaCore shows that the NF-κB signaling is the most significantly altered. (D) Similarly, the 187 genes were used in the STRING 9.1 database to visualize potential protein networks (highlighted in red are proteins of the NF-κB signaling). The confidence view is shown: stronger association between proteins is represented by thicker lines (disconnected nodes are hidden).