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Trichosanthin (TCS) is a traditional Chinese herbal medicine to treat some gynecological diseases. Its effective component has diverse biological function, including antineoplastic activity. The human trophoblast cell line BeWo was chosen as an experimental model for in vitro test of drug screen for anti-cancer property of TCS. MTT method was used in this study to get primary screen result. The result showed that 100 mM had the best IC50 value. Proteomics analysis was then performed for further investigation for the drug effect of TCS to the BeWo cell line. In this differential proteomic expression analysis, total protein extract from BeWo cell line and their protein expression level after the drug treatment were compared by 2DE. Then 24 unique three-fold differential expressed proteins (DEPs) were successfully identified by MALDI-TOF-TOF-MS. Label-free proteomics as complemental method was run for the same experimental procedure. There are two proteins were identified in both 2DE and label free methods. Among those identified proteins, bioinformatics analysis shows the importance of pathway and signal transduction and gives us the potential possibility for the disease treatment hypothesis.