Comparative proteomic analysis of beta-catenin-mediated malignant progression of esophageal squamous cell carcinoma.
beta-catenin has emerged as a key regulator of Wnt signaling pathway, which plays an important role in the development and progression of various cancers. Its accumulation in nucleus of the esophagus squamous epithelium might be the crucial step for the carcinogenesis of esophageal squamous cell carcinoma (ESCC). To detect the proteins correlated with beta-catenin function, we used the established cell lines of pGen-3-con (Eca109 cells transfected by control vector) and pGen-3-CTNNB1 (Eca109 cells transfected by beta-catenin siRNA) as cell models for further analysis. Two-dimensional gel electrophoresis technology was performed to separate the proteins of pGen-3-con and pGen-3-CTNNB1 cell lines, respectively. The differential protein spots were analyzed by software analysis, subjected to in-gel digestion, and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Consequently, 13 differentially expressed proteins between the two cell lines were identified, of which 14-3-3sigma, prohibitin, and nm23-H1 were further verified by western blotting and quantitative real-time reverse transcriptase-polymerase chain reaction. Then, the tissue microarray and immunohistochemical analysis were employed to research their relationship in ESCC and their corresponding normal mucosa tissues. The upregulation of prohibitin or the downregulation of 14-3-3sigma and nm23-H1 proteins was significantly associated with the proliferation, invasion depth, and lymph node metastasis of ESCC. There were statistically significant correlations between the expression of beta-catenin and the three proteins. The results presented here might provide potential protein markers to elucidate the mechanism of beta-catenin-mediated biologic characteristics for ESCC.
Ren
H-Z
HZ
Department of Pathology, Xiangya Medical College, Ministry of Health of China, Xiangya Hospital, Central South University, Changsha 410078, China.
Wang
J-S
JS
Pan
G-Q
GQ
Lv
H
H
Wen
J-F
JF
Luo
G-Q
GQ
Wang
K-S
KS
Zhang
P-F
PF
D048948:14-3-3 Proteins / Q000032:analysis / Q000235:genetics; D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018450:Disease Progression; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D004938:Esophageal Neoplasms / Q000235:genetics / Q000473:pathology*; D004947:Esophagus / Q000166:cytology; D005092:Exonucleases / Q000032:analysis / Q000235:genetics; D005095:Exoribonucleases; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D008207:Lymphatic Metastasis / Q000473:pathology; D054778:NM23 Nucleoside Diphosphate Kinases / Q000032:analysis / Q000235:genetics; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000032:analysis / Q000235:genetics; D000091402:Prohibitins; D040081:Protein Array Analysis; D020543:Proteome / Q000032:analysis*; D012097:Repressor Proteins / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014162:Transfection; D015854:Up-Regulation; D051176:beta Catenin / Q000032:analysis* / Q000235:genetics
D048948:14-3-3 Proteins; D014408:Biomarkers, Tumor; C495270:CTNNB1 protein, human; D054778:NM23 Nucleoside Diphosphate Kinases; D009363:Neoplasm Proteins; D000091402:Prohibitins; D020543:Proteome; D012097:Repressor Proteins; D051176:beta Catenin; C517938:NME1 protein, human; D005092:Exonucleases; D005095:Exoribonucleases; C413442:SFN protein, human