Identification of Rack1, EF-Tu and Rhodanese as aging-related proteins in human colonic epithelium by proteomic analysis.
The aging process of human colonic epithelium involves a slow decline in physiological vigor and an increasing susceptibility to age-related diseases, especially, colon cancer, but the mechanisms still remain to be elucidated. To reveal the molecular bases of colonic epithelial aging, a proteomic approach was used to screen for differential proteins in the human normal colonic epithelial tissues from young and old people. As a result, 17 differential proteins were identified by two-dimensional electrophoresis and mass spectrometry, and the partial differential proteins were confirmed by immunohistochemistry. Rack1, EF-Tu and Rhodanese, three validated differential proteins, were further investigated for their role in the in vitro cell senescence. Western blot showed that the expression of all the three proteins was downregulated in the senescent NIH/3T3 cells induced by D-galactose as compared to the control cells. Furthermore, knockdown of Rack1 by siRNA could promote NIH/3T3 cell senescence. Taken together, our results suggest that Rack1, EF-Tu and Rhodanese are aging-related proteins in human colonic epithelium, and injury of mitochondrial function and decline of antioxidant capability are important reasons for the aging of human colonic epithelium.
Yi
Hong
H
Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
Li
Xin-Hui
XH
Yi
Bin
B
Zheng
Jie
J
Zhu
Guo
G
Li
Cui
C
Li
Mao-Yu
MY
Zhang
Peng-Fei
PF
Li
Jian-Ling
JL
Chen
Zhu-Chu
ZC
Xiao
Zhi-Qiang
ZQ
D000368:Aged; D000818:Animals; D015153:Blotting, Western; D016922:Cellular Senescence / Q000187:drug effects / Q000502:physiology*; D003106:Colon / Q000166:cytology; D015180:Electrophoresis, Gel, Two-Dimensional; D019204:GTP-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D005690:Galactose / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D041681:NIH 3T3 Cells; D009363:Neoplasm Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D010444:Peptide Elongation Factor Tu / Q000032:analysis / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D034741:RNA, Small Interfering; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D013884:Thiosulfate Sulfurtransferase / Q000032:analysis / Q000378:metabolism*; D055815:Young Adult
D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011506:Proteins; C419368:RACK1 protein, human; D034741:RNA, Small Interfering; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D013884:Thiosulfate Sulfurtransferase; D019204:GTP-Binding Proteins; D010444:Peptide Elongation Factor Tu; D005690:Galactose