JIN-YUAN SHIHPAN-CHYR YANGHong T.-M.ANG YUANChen J.J.W.CHONG-JEN YUYIH-LEONG CHANGLee Y.-C.Peck K.Wu C.-W.Yang P.-C.2020-03-072020-03-0720010027-8874https://www.scopus.com/inward/record.uri?eid=2-s2.0-0035913647&doi=10.1093%2fjnci%2f93.18.1392&partnerID=40&md5=8aef2ee5fa6f6cc96df0f9bb255ca7e1https://scholars.lib.ntu.edu.tw/handle/123456789/473946Background: 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.[SDGs]SDG3cell protein; collagen response mediator proten 1; green fluorescent protein; unclassified drug; adult; advanced cancer; aged; article; assay; cancer cell; cancer cell culture; cancer genetics; cancer invasion; cancer recurrence; cancer staging; cancer survival; cell activity; cell invasion; cell nucleus; cell shape; cell structure; centrosome; controlled study; cytoplasm; DNA microarray; female; fluorescence microscopy; gene expression; gene overexpression; gene sequence; genetic transfection; human; human cell; in vitro study; interphase; lung cancer; lymph node metastasis; male; metastasis; mitosis; mitosis spindle; Northern blotting; nucleotide sequence; postoperative period; priority journal; protein analysis; protein localization; reverse transcription polymerase chain reaction; statistical analysis; suppressor gene; telophase; Western blottingjournal article10.1093/jnci/93.18.1392115623902-s2.0-0035913647