Objective. The objective of the present study is to evaluate and compare the infrared spectral features of normal and malignant exfoliated cervical cells, cells from malignant tissue, and the SiSo cell line.Methods. Infrared spectra of cervical adenocarcinoma (CA) tissue, normal and malignant exfoliated cervical cells, and a uterine cervical adenocarcinoma cell line (SiSo) were obtained. Spectral qualities in terms of band intensity ratio and band position, which reflect configurational changes in the functional groups of the above samples, were measured.Results. Spectral bands of CA tissue, exfoliated cells from CA, and the cell line were similar but markedly different from that of exfoliated normal cervical cells. Significant changes in bands at 1025 cm−1 (glycogen), 1080 cm−1 (glycogen and nucleic acids), 1155 cm−1 (C–OH groups of serine, threonine, and tyrosine of cell proteins, and C–O groups of carbohydrates), 1240 cm−1 (PO2 groups of nucleic acids), 1400 cm−1 (methyl group of lipids and proteins), and 1450 cm−1 (methylene group of lipids and proteins) were noted in the CA tissue, exfoliated CA cells, and adenocarcinoma cell line compared with exfoliated normal cells. Marked shifts in band positions from 1080 to 1086 cm−1, 1153 to 1160 cm−1, and 935 to 970 cm−1 in CA tissue, exfoliated CA cells, and the adenocarcinoma cell line were noted.Conclusion. Spectral bands of the adenocarcinoma cell line matched very well with those of cervical CA tissue and exfoliated CA cells in terms of position. In contrast, spectral bands of the SiSo cell line differed greatly from those of normal exfoliated cells.