Matriptase is an epithelial-derived, integral serine protease that has been implicated in the progression of epithelial tumors. We investigated whether the expression of matriptase is associated with the progression of cervical neoplasia. Using immunohistochemistry, we evaluated the matriptase expression in 89 formalin-fixed paraffin-embedded cervical tissues that included 10 normal cervical specimens, 19 low-grade squamous intraepithelial lesions, 20 high-grade squamous intraepithelial lesions, 20 invasive squamous cell carcinomas (ISCC) without lymph node (LN) metastasis, and 20 ISCC with lymph node metastasis. We also used the reverse transcriptase-polymerase chain reaction technique to determine the expression of matriptase transcripts in normal cervical and ISCC tissues. The immunohistochemical staining showed that the expression of matriptase was undetectable in all normal cervical squamous epithelia, but had cytoplasmic and membranous staining in the normal endocervical glands. Staining gradually increased in accordance with the histopathologic grades from low-grade squamous intraepithelial lesions to high-grade squamous intraepithelial lesions and ISCC ( P < .001); matriptase was detected in most cases (95%) of ISCC. In addition, matriptase transcripts were expressed in all (n = 26) of the ISCC cases by microdissection and reverse transcriptase-polymerase chain reaction, whereas none of the normal squamous epithelia cases (n = 3) expressed matriptase transcripts. These results suggest that matriptase may play a significant role in the development of cervical carcinoma and may serve as a useful marker of the malignant transformation of cervical squamous cells. Further studies could potentially lead to the development of novel approaches for early detection and therapy for this disease.