Raman microspectroscopy can provide molecular-level finger-print information about the biochemical composition and structure of cells and tissues with excellent spatial resolution. Raman spectroscopy of nasopharyngeal carcinoma cell lines C666-1, CNE2 and normal nasopharyngeal cell line NP69 are presented to investigate the differences between them. The ratio of I1449/I1657(1.10) seems to very easily divide tumor and normal cell lines into two groups, and this result is coincident with the existing report about the study of normal and malignant bronchial tissue. Principal component analysis (PCA) and linear discriminant analysis (LDA) are also used to classify different cell lines and achieve an exciting result with a sensitivity and specificity of 90% and 100%, respectively. The results of the work may be helpful to the discrimination of normal and tumor cells, which show that Raman spectroscopy can be one of the diagnostic methods of nasopharyngeal carcinoma and do favor to the early diagnosis.