Studies On The Anti-Tumor Effects Of Proteins From Trachyrhamphus Serratus In Vitro

In view of the need for new anti-cancer compounds with low toxic potential, numerous proteins from different Chinese Traditional Medicine have been investigated for anti-tumor and immunomodulating activities. In this thesis, the anti-tumor activity of the water-soluble proteins (HCP) from Trachyrhamphus serratus was studied.After the tumor cell lines were cultured with 0.008、0.04、0.2、1、5 mg/ml HCP in vitro,the proliferation were evaluated by MTT assay. The results showed HCP had anti-proliferation effect on tumor cell lines L1210, CCRF-CEM, A549 and LOVO, and the inhibition influences on cell lines were directly proportioned to the HCP concentrations in the range of 0.008～5 mg/ml.Cell cycle, and apoptosis of A549 cells were analyzed by flow cytometry; apoptosis induction was also proved by electrophoresis and morphologic assessment. As a result, the crude proteins induced apoptosis in A549 cells. In this thesis, an anti-tumor active protein, namely S1, was purified with the methods of DEAE-Sepharose Fast Flow column chromatogramphy and Superdex 75 prep grade chromatography. S1 has main bands with 58.3 kDa by SDS-PAGE, the isoelectric point is about 4.4. Furthermore, the protein could inhibit the growth of tumor cell lines CCRF-CEM, L1210 and A549 in vitro with IC500.44±0.04 mg/ml, 0.32±0.06 mg/ml and 0.28±0.09 mg/ml respectively.