Primary pulmonary myxoid sarcoma (PPMS) is a very rare lung tumor that has recently been shown to harbor an EWSR1-CREB1 translocation. However, the histogenesis and biological behavior of PPMS remains unclear. To provide insight into the histogenesis of PPMS, we studied surgical resection specimens of four patients, two females and two males with an age range of 26 to 65 years, all non-smokers with mild anemia. The tumors, three of which are endobronchial, measured between 4 and 13 cm. One patient developed metastasis to the contra-lateral lung 7 months after resection. Other patients remained alive without tumor for 1.5, 10, and 13 years. Fluorescence in situ hybridization (FISH) analysis with a gene break apart probe showed an EWSR1 translocation in all cases. The EWSR1-CREB1 fusion transcript was detected in all cases by reverse-transcription PCR. Immunohistochemical staining showed diffuse positive staining of the tumor cells only for vimentin. Tumor cells expressed no other myoid, epithelial, endothelial, melanocytic, myoepithelial, or neuroendocrine markers, except for smooth muscle actin and epithelial membrane antigen, which were only focally positive in individual cases. Ultrastructural analyses revealed the presence in the tumor cells of intermediate filaments with focal densities along the sub-cytoplasmic membrane as well as dense plaques. These results suggest that PPMS exhibits myofibroblastic differentiation. We conclude that PPMS is an intermediate grade malignant lung tumor harboring EWSR1 translocations, which may originate from mesenchymal cells that undergo fibroblastic or myofibroblastic differentiation.