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摘要
[Objective]The objective of this experiment is to investigate proteomic profile of HT29 treated with high-dosage-violacein and low-dosage-violacein, and to analyze its involved metabolic pathway and mechanism. [Method]Cytotoxic activity of violacein was analyzed by MTT assay and observed by using transmission electron microscopy. After that, the total protein was extracted from the cells.To obtain relative abundance of peptides information, total proteins labeled with multiple iTRAQ stable isotopes were segregated and analyzed with RP-LC-MS/MS. The differential expression proteins were identified through NCBI. Human. Protein database. The metabolic pathway was analyzed through Go analysis and KEGG analysis. [Result]Violacein inhibited the growth of HT29 in dose-dependent and time-dependent manner. Compared with 5-Flu, 5 mg·L-1 violacein resulted in 50％ inhibition of HT29. Its dosage was one sixth of the dosage of 5-Fu. With the increasing violacein, vacuoles in the mitochondrion and membrane blebs were found under transmission electron micrograph. Violet pigment at 30 mg·L-1 induced margination of nuclear chromatin and disappearance of cell membrane. Through quantitative proteomic analysis of these three cells, 4258 proteins were identified with MS, 757 of which were differential expression proteins (fold change of protein expression≥2). Among these differential expression proteins, there were 492 proteins in cells treated with high dosage of violacein, 112 proteins in cells treated with low dosage of violacein, and 336 proteins in cells treated with 5-Fu. Analysis of enriched KEGG pathway showed that most of these differential expression proteins were involved in 50 signaling pathways, 10 of which were enriched significantly (P <0.05), such as ribosome, citrate cycle and RNA degradation. [Conclusion]The antitumor mechanism of violacein against colon cancer cells are mainly involved in transcription and translation in cell life cycle.
- 出版日期2017
- 单位中国农业科学院
