The fms oncogene encodes the macrophage colony-stimulating factor receptor (CSF1R), a transmembrane tyrosine kinase receptor, which is abnormally expressed in breast cancer. Transfection of wild-type CSF1R into HC11 mammary epithelial cells (HC11-CSF1R) renders the transfectants capable of in vitro local invasion and in vivo tumorigenesis. Transfection with CSF1R mutated to express phe at the tyr-721 autophosphorylation site (HC11-CSF1R-721) creates a phenotype that lacks metastastic competence but maintains local invasiveness. Conversely, HC11 cells transfected with CSF1R mutated at tyr-807 (HC11-CSF1R-807) retain their metastatic competence, but are not locally invasive. Our aims were to determine which genes were differentially expressed with transfection of HC11 with wild-type CSF1R, and to determine the effect of mutation at the autophosphorylation sites on gene expression, using 4.6 K cDNA microarrays. Complementary DNA from HC11, HC11-CSF1R-721 and HC11-CSF1R-807 were each hybridized together with HC11-CSF1R on individual arrays. A principal component spectral method combined with prenormalization procedures was used for sample clustering. Differentially expressed genes were identified by the analysis of variance. Confirmation by Northern blotting was performed for MAP kinase phosphatase-1, WDNM1 (extracellular proteinase inhibitor), Trop 2 (tumor-associated calcium signal transducer-2), procollagen type IV alpha, secretory leukoprotease inhibitor, prenylated snare protein Ykt6, ceruloplasmin and chaperonin 10. Many of these genes have not previously been associated with tumor invasion and metastasis. We have successfully identified genes that can be linked to the invasive phenotypes or to tumorigenesis. These genes provide a basis for further studies of metastatic progression and local invasiveness, and can be evaluated as therapeutic targets. The cfms proto-oncogene encodes the only known receptor for the macrophage colony-stimulating factor (CSF1). CSF1R is a transmembrane tyrosine kinase receptor, and its ligand, CSF1, has soluble, membrane-bound and cell matrix-associated isoforms. 1-3 The CSF1R/CSF1 receptor/ligand pair has essential physiologic functions in monocyte and macrophage differentiation, 4,5 embryonic implantation and placental development, and lactogenic differentiation of the human breast. 6-8 Abnormally high CSF1R expression has clinically been associated with aggressive breast, ovarian, endometrial, and prostate cancer. [9][10][11][12][13][14][15][16] Most invasive breast carcinoma cells express readily detectable levels of activated CSF1R. 13 Studies 17 have shown that invasive breast cancer cells coexpress the ligand CSF 1, but adjacent in situ carcinoma cells do not. Moreover, in early-stage breast cancer patients treated with local therapy only, high levels of CSF1R expression also have been associated with a higher likelihood of ipsilateral recurrence. 18 To further evaluate the role of CSF1R in invasion and metastasis, we studied a mouse cell line model 19 that utilized the HC11...