RS is a well-established quantifiable optical technique with excellent reproducibility, and its spectrum might provide information in identifying structural changes of molecule. Each molecule had its own pattern of vibration that could serve as a “Raman biomarker.”18 RS had been used to accurately identify benign prostate hyperplasia and PC tissues, otherwise, heterogenetic cell lines (LNCaP, CaP2b, PC-3, and DU-145) with significant biochemical differences were applied in their research,19,29 but whether RS could identify congelleric CRPC and ADPC cells and tissues in vivo was still confusing. We used the classic LNCaP/C4-2 cells as cell model which mimicked closely the natural history from ADPC to CRPC (20), then selected and matched the PC samples with coincident Gleason score to reduce the intrinsic interference. Notably, spectra of the nine paired patients presented more reliable results in vitro. We compared the spectral changes between CRPC and ADPC in both cell and tissue models. It revealed the corresponding spectral changes reflected structural changes of protein or DNA, concomitant with the alteration of amino acid and basic radical. Depending on these changes of conformation or component in some biomolecules, PCA/SVM effectively identified CRPC and ADPC. However, these spectral changes only represented partial information of the biomacromolecules, such as the level of secondary protein structure. Further, molecular biological studies were needed to determine the mechanism of these alterations.