Abstract: This study aims to establish a carcinogenesis model of human squamous tongue cancer cell in vitro, and test by laser confocal micro Raman spectrometer, then analyze from the molecular level.MethodsThe CAL-27 cells were cultivated, and injected subcutaneously in nude mice into a tumor, then using laser confocal micro Raman spectrometer and HE staining to observe the canceration.Results①From the normal skin tissue slice, it can be seen that the cellular level is clear and neat. But it can be found that the tumor epithelial dysplasia is cellular pleomorphism and the structure level is not clear.②After laser confocal micro Raman spectrometerobsersving, the principal components analysis(PCA) combination with linear discrimination analysis(LDA) to classify the health nude mice and human tongue squamous cancer cells in vitro tumor tissue. It can be obtained the diagnostic sensitivity of 98.4%,specificity of 95% and accuracies of 96.7%.It was shown that this method is very good for detecting the animal modelin vivo.