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Profiles of metabolic protein expressions in proteomic and phospho‐proteomic analysis of low folate modulates energy metabolism bioenergetics in colon cancer
Author(s) -
Lan WenYu,
Ku WeiChi,
Huang RweiFen Syu.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.802.27
Subject(s) - biochemistry , proteomics , chemistry , bioenergetics , metabolism , cancer cell , biology , cancer , mitochondrion , genetics , gene
Previous studies we found that low folate microenvironment promotes cancer stem cell properties. Induction of cancer stem cells is associated with energy metabolism. How folate nutritional status induces cancer stem cell‐like potentials and energy switch mechanisms remains unclear. Aims of this study were to investigate profiles of metabolic protein expressions in proteomic and phospho‐proteomic analysis of low folate modulates energy metabolism bioenergetics in colon cancer. Human colon cancer cell line SW480 were cultured in medium containing 2.2 μM folate (control (C) group), 10 nM folate (low folate (LF) group) 4 days and 8 days. SW480 cells (control and LF) were scraped in a lysis buffer containing 4% SDS and protease inhibitor cocktails. After acetone precipitation, total proteins were digested by trypsin followed by dimethyl labeling at the peptide level. The labeled peptides were combined and analyzed by nanoLC‐MS/MS. The peptides were separated by MonoCap C18 High Resolution Ultra 2000 (0.1 × 2000 mm) column and analyzed by Thermo LTQ‐Orbitrap XL mass spectrometer. Peptide identification and quantification were performed by MaxQuant packages. Bio‐informatics analyses were performed by Perseus and GproX program. These proteomic analysis data observed regulated proteins related to energy metabolism, lipid metabolism, and TOR signaling. Among these proteins, mTOR, pyruvate dehydrogenase‐phosphatase 1, ADP/ATP translocase 3 were over 2 fold‐changes in LF group. ATP synthase was under 2 fold‐changes in LF group. Using phospho‐proteomic analysis data revealed that MAPK1, IRS2 were over 2 fold‐changes in LF group. Akt, S6K1, and pyruvate dehydrogenase protein phosphorylation were significantly up‐regulated in LF group. Overall protein and phosphorylated proteins related to metabolism were overlap and associated with cancer stem cell‐like potentials. And there were onco‐spheroid formation cells in LF treated group. These findings demonstrated that proteomic analysis was applied to metabolic protein expressions of low folate modulates energy metabolism bioenergetics in colon cancer.