Identification of differential proteins in colorectal cancer cells treated with caffeic acid phenethyl ester.

Abstract

AIM:

To investigate the molecular mechanisms of the anti-cancer activity of caffeic acid phenethyl ester (CAPE).

METHODS:

Protein profiles of human colorectal cancer SW480 cells treated with or without CAPE were analysed using a two-dimensional (2D) electrophoresis gel-based proteomics approach. After electrophoresis, the gels were stained with Coomassie brilliant blue R-250. Digital images were taken with a GS-800 Calibrated Densitometer, and image analysis was performed using PDQuest 2-D Analysis software. The altered proteins following CAPE treatment were further identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry following a database search. The identified proteins were validated by Western blot and immunofluorescence assay.

RESULTS:

CAPE induced human colorectal cancer cell apoptosis. Four up-regulated proteins and seven down-regulated proteins in colorectal cancer cells treated with CAPE were found. The identified down-regulated proteins in CAPE-treated colorectal cancer cells were Triosephosphate Isomerase (Tim), Proteasome subunit alpha 4 (PSMA4) protein, Guanine nucleotide binding protein beta, Phosphoserine aminotransferase 1 (PSAT1), PSMA1, Myosin XVIIIB and Tryptophanyl-tRNA synthetase. Notably, CAPE treatment led to the down-regulation of PSAT1 and PSMA1, two proteins that have been implicated in tumorigenesis. The identified up-regulated proteins were Annexin A4, glyceraldehyde-3-phosphate dehydrogenase, Glucosamine-6-phosphate deaminase 1 (GNPDA1), and Glutathione peroxidase (GPX-1). Based on high match scores and potential role in cell growth control, PSMA1, PSAT1, GNPDA1 and GPX-1 were further validated by Western blotting and immunofluorescence assay. PSMA1 and PSAT1 were down-regulated, while GNPDA1 and GPX-1 were up-regulated in CAPE-treated colorectal cancer cells.

CONCLUSION:

These differentiated proteins in colorectal cancer cells following CAPE treatment, may be potential molecular targets of CAPE and involved in the anti-cancer effect of CAPE.

• Authors: Yu-Jun He, Wan-Ling Li, Bao-Hua Liu, Hui Dong, Zhi-Rong Mou, Yu-Zhang Wu
• Journal: World journal of gastroenterology (World J Gastroenterol) Vol. 20 Issue 33 Pg. 11840-9 (Sep 07 2014) ISSN: 2219-2840 [Electronic] United States
• PMID: 25206290 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents
• Biomarkers, Tumor
• Caffeic Acids
• Neoplasm Proteins
• caffeic acid phenethyl ester
• Phenylethyl Alcohol

Topics

• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects)
• Biomarkers, Tumor (metabolism)
• Blotting, Western
• Caffeic Acids (pharmacology)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Colorectal Neoplasms (metabolism, pathology)
• Electrophoresis, Gel, Two-Dimensional
• Fluorescent Antibody Technique
• Humans
• Microscopy, Confocal
• Neoplasm Proteins (metabolism)
• Phenylethyl Alcohol (analogs & derivatives, pharmacology)
• Proteomics (methods)
• Reproducibility of Results
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization