Tumor cell migration screen identifies SRPK1 as breast cancer metastasis determinant

J Clin Invest. 2015 Apr;125(4):1648-64. doi: 10.1172/JCI74440. Epub 2015 Mar 16.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Bone Neoplasms / secondary
  • Breast Neoplasms / genetics*
  • Carcinoma, Non-Small-Cell Lung / pathology
  • Cell Adhesion
  • Cell Movement / genetics
  • Cell Polarity
  • Female
  • Focal Adhesions / physiology
  • Gene Expression Regulation, Neoplastic
  • Genetic Association Studies
  • Humans
  • Kaplan-Meier Estimate
  • Lung Neoplasms / pathology
  • Lung Neoplasms / secondary
  • Mice
  • NF-kappa B / metabolism
  • Neoplasm Metastasis / genetics*
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology*
  • Nuclear Proteins / physiology
  • Organ Specificity
  • Prognosis
  • Protein-Serine-Threonine Kinases / deficiency
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / physiology*
  • RNA Interference
  • RNA, Small Interfering / genetics

Substances

  • ITGB3BP protein, human
  • NF-kappa B
  • Neoplasm Proteins
  • Nuclear Proteins
  • RNA, Small Interfering
  • SRPK1 protein, human
  • Srpk1 protein, mouse
  • Protein-Serine-Threonine Kinases