© 2001 by Oxford University Press
Journal of the National Cancer Institute, Vol. 93, No. 18, 1392-1400,
September 19, 2001
© 2001 Oxford University Press
REPORT |
Collapsin Response Mediator Protein-1 and the Invasion and Metastasis of Cancer Cells
Affiliations of authors: J.-Y. Shih, C.-J. Yu (Department of Internal Medicine), A. Yuan (Department of Emergency Medicine), J. J. W. Chen (Department of Clinical Research), Y.-L. Chang (Department of Pathology), Y.-C. Lee (Department of Surgery), National Taiwan University Hospital, Taipei; S.-C. Yang, T.-M. Hong, K. Peck, Institute of Biomedical Sciences, Academia Sinica, Taipei; C.-W. Wu, National Health Research Institutes, Taipei; P.-C. Yang, Department of Internal Medicine, National Taiwan University Hospital, National Health Research Institutes, and Institute of Biomedical Sciences, Taipei.
Correspondence to: Pan-Chyr Yang, M.D., Ph.D., Department of Internal Medicine, National Taiwan University Hospital, No. 7, Chung-Shan South Rd., Taipei 100, Taiwan (e-mail: pcyang{at}ha.mc.ntu.edu.tw).
Background: Numerous genetic changes are associated with metastasis and invasion of cancer cells. To identify differentially expressed invasion-associated genes, we screened a panel of lung cancer cell lines (CL1–0, CL1–1, CL1–5, and CL1–5-F4 in order of increasing invasive activity) for such genes and selected one gene, collapsin response mediator protein-1 (CRMP-1), to characterize. Methods: We used a microarray containing 9600 gene sequences to assess gene expression in the cell panel and selected the differentially expressed CRMP-1 gene for further study. We confirmed the differential expression of CRMP-1 with northern and western blot analyses. After transfecting and overexpressing CRMP-1 in highly invasive CL1–5 cells, the cells were assessed morphologically and with an in vitro invasion assay. We used enhanced green fluorescent protein-tagged CRMP-1 and fluorescence microscopy to localize CRMP-1 intracellularly. CRMP-1 expression in 80 lung cancer specimens was determined by real-time quantitative reverse transcription–polymerase chain reaction (RT–PCR). All statistical tests were two-sided. Results: Expression of CRMP-1 was inversely associated with invasive activity in the cell panel, an observation confirmed by northern and western blot analyses. CRMP-1-transfected CL1–5 cells became rounded and had fewer filopodia and statistically significantly lower in vitro invasive activity than untransfected cells (all P<.001). During interphase, CRMP-1 protein was present uniformly throughout the cytoplasm and sometimes in the nucleus; during mitosis, CRMP-1 was associated with mitotic spindles, centrosomes, and the midbody (in late telophase). Real-time RT-PCR of lung cancer specimens showed that reduced expression of CRMP-1 was statistically significantly associated with advanced disease (stage III or IV; P = .010), lymph node metastasis (N1, N2, and N3; P = .043), early postoperative relapse (P = .030), and shorter survival (P = .016). Conclusions: CRMP-1 appears to be involved in cancer invasion and metastasis and may be an invasion-suppressor gene.
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