TY - JOUR
T1 - A novel polypeptide extracted from Ciona savignyi induces apoptosis through a mitochondrial-mediated pathway in human colorectal carcinoma cells
AU - Cheng, Linyou
AU - Wang, Chunguang
AU - Liu, Haizhou
AU - Wang, Fengxia
AU - Zheng, Lanhong
AU - Zhao, Jin
AU - Chu, Edward
AU - Lin, Xiukun
N1 - Funding Information:
This study was supported by 863 High Technology Project (No. 2007AA091403 and 2007AA09Z408 from the Chinese Ministry of Science and Innovative Drug Development Projects of China (2009ZX09103-661 and 2009ZX09102). We are grateful to all members of the laboratory for their continuous technical advice and helpful discussion.
PY - 2012/9
Y1 - 2012/9
N2 - Sessile marine animals such as sponges, ascidians, and bryozoans are a rich source of bioactive natural products, many of which exhibit potent anticancer activity. We extracted and purified a polypeptide with potent antitumor activity from Ciona savignyi by acetone fractionation, ultrafiltration, ion exchange chromatography, gel chromatography, and high-performance liquid chromatography. An MTT assay was used to study the cytotoxicity of the isolated fraction and the purified polypeptide. Cell cycle and Western blot analysis were performed to study the mode of action of the purified polypeptide. A novel polypeptide with potent antitumor activity was purified. The molecular weight of the polypeptide, designated CS5931, was 5931 Da, and use of the genome basic local alignment search tool (BLAST) revealed that the N-terminal sequence of CS5931 is identical to that of granulin A from C savignyi. CS5931 exhibited significant cytotoxicity for several cancer cell types and induced apoptotic death in HCT-8 cells in a dose- and time-dependent manner. Cell cycle analysis demonstrated that CS5931 caused cell cycle arrest at the G2/M phase, and a sub-G1 peak appeared after treating the cells with CS5931 for 12 hours. The mitochondrial-mediated pathway was implicated in CS5931-induced apoptosis. Our observations clearly demonstrate the antiproliferative and proapoptotic activities of the polypeptide CS5931 from C savignyi and the mitochondrial-mediated pathway involved in the polypeptide-induced cell death.
AB - Sessile marine animals such as sponges, ascidians, and bryozoans are a rich source of bioactive natural products, many of which exhibit potent anticancer activity. We extracted and purified a polypeptide with potent antitumor activity from Ciona savignyi by acetone fractionation, ultrafiltration, ion exchange chromatography, gel chromatography, and high-performance liquid chromatography. An MTT assay was used to study the cytotoxicity of the isolated fraction and the purified polypeptide. Cell cycle and Western blot analysis were performed to study the mode of action of the purified polypeptide. A novel polypeptide with potent antitumor activity was purified. The molecular weight of the polypeptide, designated CS5931, was 5931 Da, and use of the genome basic local alignment search tool (BLAST) revealed that the N-terminal sequence of CS5931 is identical to that of granulin A from C savignyi. CS5931 exhibited significant cytotoxicity for several cancer cell types and induced apoptotic death in HCT-8 cells in a dose- and time-dependent manner. Cell cycle analysis demonstrated that CS5931 caused cell cycle arrest at the G2/M phase, and a sub-G1 peak appeared after treating the cells with CS5931 for 12 hours. The mitochondrial-mediated pathway was implicated in CS5931-induced apoptosis. Our observations clearly demonstrate the antiproliferative and proapoptotic activities of the polypeptide CS5931 from C savignyi and the mitochondrial-mediated pathway involved in the polypeptide-induced cell death.
KW - Antitumor
KW - Ciona savignyi
KW - Polypeptide
KW - Purification and identification
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U2 - 10.1016/j.clcc.2012.01.002
DO - 10.1016/j.clcc.2012.01.002
M3 - Article
C2 - 22440403
AN - SCOPUS:84865504525
SN - 1533-0028
VL - 11
SP - 207
EP - 214
JO - Clinical colorectal cancer
JF - Clinical colorectal cancer
IS - 3
ER -