Abstract：Objective To identify the differential proteins in gallbladder cancer group and normal gallbladder tissue control group, and find and identify the tumor markers related to gallbladder cancer and potential therapeutic targets by using classical proteomic techniques, two-dimensional gel electrophoresis, mass spectrometry identification and bioinformatics analysis. Methods The samples were divided into two groups: gallbladder cancer group (59 cases) and normal control group (40 cases). The total protein samples were collected by washing, lysis and centrifugation of the tissue samples, and then packaged and frozen. After the first direction isofocal focusing and the second SDS-PAGE electrophoresis, the gel was dyed and colored with Coomnassie blue, and the gel image was obtained by Gs-800 optical densitometer. After analysis by PD Quest gel image analysis software, different protein spots were found. Secondary mass spectrometry of differential protein fragments was obtained using Peptide Mass Fingerprinting (PMF) and Matrix Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) mass spectrometry techniques. Biotools software was used to collect mass- charge ratio information of peptide fragments. The Mascot search engine was used to search the database, identify the amino acid sequences of differential proteins, and obtain the corresponding specific protein information. Results A total of 66 differentially expressed proteins were found, Among which, 21 proteins were up-regulated and 45 proteins were down-regulated. Of 21 differential proteins with scores higher than 56, 10 were up-regulated and 11 were down-regulated in gallbladder cancer. It mainly includes annexinsANX, moesin, heat shock protein (HSP), vimentin, Retinol dehydrogenase 1, galectin, and so on. Conclusions The above-mentioned proteins may play an important role in the occurrence and development mechanism of tumors, and are potential targets for diagnosis and treatment, which deserves further research in the future.
Morris R M,Fung J M,Rahm B G,et al.Comparative proteomics of Dehalococcoides sppreveals strain-specific peptides associated with activity[J].Appl Environ Microbio,2007,73(1): 320-326.
[1]
Morris R M,Fung J M,Rahm B G,et al.Comparative proteomics of Dehalococcoides sppreveals strain-specific peptides associated with activity[J].Appl Environ Microbio,2007,73(1): 320-326.
[2]
Ziv T,Barnea E,Segal H,et al.Comparative proteomics of small cell lung carcinoma[J]. Cancer Biomark,2006, 2(6): 219-234.
[2]
Ziv T,Barnea E,Segal H,et al.Comparative proteomics of small cell lung carcinoma[J]. Cancer Biomark,2006, 2(6): 219-234.
[3]
Lisacek F, Cohen B S,Appel R D.Proteome informatics II: bioinformatics for comparative proteomics[J].Proteomics,2006, 6(20): 5445-566.
[3]
Lisacek F, Cohen B S,Appel R D.Proteome informatics II: bioinformatics for comparative proteomics[J].Proteomics,2006, 6(20): 5445-566.
[4]
Naaby H S,Waterfield M D,Cramer R.Proteomics--post-genomic cartography to understand gene function[J].Trends Pharmacol Sci,2001,22(7): 376-384.
[4]
Naaby H S,Waterfield M D,Cramer R.Proteomics--post-genomic cartography to understand gene function[J].Trends Pharmacol Sci,2001,22(7): 376-384.
[5]
Cormican J A, Soh W T, Mishto M, et al.iBench: a ground truth approach for advanced validation of mass spectrometry identification method[J].Proteomics,2023,23(2):2200271.
[5]
Cormican J A, Soh W T, Mishto M, et al.iBench: a ground truth approach for advanced validation of mass spectrometry identification method[J].Proteomics,2023,23(2):2200271.
[6]
Wasinger V C,Cordwell S J,Cerpa P A,et al.Progress with gene product mapping of the Mdlicutos:Mycoplaanla genitalium[J].Electrophoresis,1995,16(7):1090-1094.
[6]
Wasinger V C,Cordwell S J,Cerpa P A,et al.Progress with gene product mapping of the Mdlicutos:Mycoplaanla genitalium[J].Electrophoresis,1995,16(7):1090-1094.
[7]
Rahman R R,Roblick U J,Hellman U,et al.p53 targets by protein expression profiling[J].Proc Natl Acad Sci USA,2007,104(13): 5401-5406.
[7]
Rahman R R,Roblick U J,Hellman U,et al.p53 targets by protein expression profiling[J].Proc Natl Acad Sci USA,2007,104(13): 5401-5406.
[8]
Cho W C.Contribution of oncoproteomics to cancer biomarker discovery[J].Mol Cancer,2007,6: 25.
[8]
Cho W C.Contribution of oncoproteomics to cancer biomarker discovery[J].Mol Cancer,2007,6: 25.
[9]
Chung C H,Levy S,Chaurand P,et al.Genomics and proteomics: emerging technologies in clinical cancer research[J].Crit Rev Oncol Hematol,2007,61(1):1-25.
[10]
O’Farell P H.High resolution two-dimensional electrephoresis of protein[J]. Biol Chem,1975,250(10):4007-4021.
[9]
Chung C H,Levy S,Chaurand P,et al.Genomics and proteomics: emerging technologies in clinical cancer research[J].Crit Rev Oncol Hematol,2007,61(1):1-25.
[10]
O’Farell P H.High resolution two-dimensional electrephoresis of protein[J]. Biol Chem,1975,250(10):4007-4021.
[11]
Klose J.Protein mapping by combined isoclectric focusing and electro-phornsis of mouse tissues.A novel approach to testing for induced pointmutations in mammals[J].Humangenetik,1975,26(3):231-243.
[11]
Klose J.Protein mapping by combined isoclectric focusing and electro-phornsis of mouse tissues.A novel approach to testing for induced pointmutations in mammals[J].Humangenetik,1975,26(3):231-243.
[12]
Calvo K R,Liotta L A, Petricoin E F. Clinical pmteomics:from biomarker discorvery and cell signaling profiles to indifidualized personal therapy[J].Bio Qei Pep,2005,25(1-2):107-125.
[13]
Gunawardana C G,Diamandis E P.High throughput proteomic strategies for identifying tumour-associated antigens[J]. Cancer Lett,2007,249(1): 110-119.
[14]
Roboz J. Mass spectrometry in diagnostic oncoproteomics[J].Cancer Invest,2005,23(5): 465-478.
[12]
Calvo K R,Liotta L A, Petricoin E F. Clinical pmteomics:from biomarker discorvery and cell signaling profiles to indifidualized personal therapy[J].Bio Qei Pep,2005,25(1-2):107-125.
[15]
Koeril M.Mass spectrometry in Proteomic and oncoproteomics[J]. Cancer,2004,16(6): 245-249.
[13]
Gunawardana C G,Diamandis E P.High throughput proteomic strategies for identifying tumour-associated antigens[J]. Cancer Lett,2007,249(1): 110-119.
[16]
Alfonso P,Canamero M,Fernandez Carbonie F, et al.Roteome analysis of membrane fractions in colorectal carcinomas by using 2D-DIGE saturation labeling[J].J Proteome Res,2008,7(10): 4247-4255.
[14]
Roboz J. Mass spectrometry in diagnostic oncoproteomics[J].Cancer Invest,2005,23(5): 465-478.
Koeril M.Mass spectrometry in Proteomic and oncoproteomics[J]. Cancer,2004,16(6): 245-249.
[18]
Hirsh M I,Junger W G.Roles of heat shock proteins and gamma delta T cells in inflammation[J].Am J Respir Cell Mol Biol,2008,39(5): 509-513.
[16]
Alfonso P,Canamero M,Fernandez Carbonie F, et al.Roteome analysis of membrane fractions in colorectal carcinomas by using 2D-DIGE saturation labeling[J].J Proteome Res,2008,7(10): 4247-4255.
[19]
Ai J H,Yang Z, Qiu F Z.Role of heat shock protein 90 in hyperdynamic circulation of portal hypertensive rats[J]. Zhonghua Yi Xue Za Zhi,2004,84(1): 9-13.
Ko S H,Suh S H,Kim B J,et al.Expression of the intermediate filament vimentin in proliferating duct cells as a marker of pancreatic precursor cell[J].Pancreas,2004,28:121-128.
[18]
Hirsh M I,Junger W G.Roles of heat shock proteins and gamma delta T cells in inflammation[J].Am J Respir Cell Mol Biol,2008,39(5): 509-513.
[21]
Beachy P A,Karhadkar S S,Berman D M.Tissue repair and stem cell renewal in carcinogenessis[J].Nature,2004,432: 324-331.
[19]
Ai J H,Yang Z, Qiu F Z.Role of heat shock protein 90 in hyperdynamic circulation of portal hypertensive rats[J]. Zhonghua Yi Xue Za Zhi,2004,84(1): 9-13.
[22]
AL H M, Clarker M F.Prospective identification of tumorigenic breast cancer cell[J].Proc Natl Acad Sci USA, 2003,100: 3983-3988.
[20]
Ko S H,Suh S H,Kim B J,et al.Expression of the intermediate filament vimentin in proliferating duct cells as a marker of pancreatic precursor cell[J].Pancreas,2004,28:121-128.
[23]
Cerri D G,Rodrigues L C,Stowell S R,et al.Degeneration of dystrophic or injured skeletal muscles induces high expression of Galectin-1[J].Glycobiology,2008,18(11): 842-850.
[21]
Beachy P A,Karhadkar S S,Berman D M.Tissue repair and stem cell renewal in carcinogenessis[J].Nature,2004,432: 324-331.
[24]
Okano K,Uchida K,Nitta K, et al.Galectin-1 suppresses alpha collagen through Smad3 in renal epithelial cells[J].Cell Mol Life Sci,2008,65(20): 3304-3311.
[22]
AL H M, Clarker M F.Prospective identification of tumorigenic breast cancer cell[J].Proc Natl Acad Sci USA, 2003,100: 3983-3988.
[25]
Akazawa C,Nakamura Y,Sango K,et al.Distribution of the galectin-1 mRNA in the rat nervous system: its transient upregulation in rat facial motor neurons after facial nerve axotomy[J].Neuroscience,2004,125(1): 171-178.
[23]
Cerri D G,Rodrigues L C,Stowell S R,et al.Degeneration of dystrophic or injured skeletal muscles induces high expression of Galectin-1[J].Glycobiology,2008,18(11): 842-850.
[26]
Brule F,Califice S,Garnier F,et al.Galectin-1 accumulation in the ovary carcinoma peritumoral stroma is induced by ovary carcinoma cells and affects both cancer cell proliferation and adhesion to laminin-1 and fibronectin[J].Lab Invest,2003, 83(3): 377-386.
[24]
Okano K,Uchida K,Nitta K, et al.Galectin-1 suppresses alpha collagen through Smad3 in renal epithelial cells[J].Cell Mol Life Sci,2008,65(20): 3304-3311.
[27]
Szoke T,Kayser K,Trojan I,et al.The role of microvascularization and growth/adhesion-regulatory lectins in the prognosis of non-small cell lung cancer in stage II[J].Eur J Cardiothorac Surg,2007,31(5): 783-787.
[25]
Akazawa C,Nakamura Y,Sango K,et al.Distribution of the galectin-1 mRNA in the rat nervous system: its transient upregulation in rat facial motor neurons after facial nerve axotomy[J].Neuroscience,2004,125(1): 171-178.
[26]
Brule F,Califice S,Garnier F,et al.Galectin-1 accumulation in the ovary carcinoma peritumoral stroma is induced by ovary carcinoma cells and affects both cancer cell proliferation and adhesion to laminin-1 and fibronectin[J].Lab Invest,2003, 83(3): 377-386.
[27]
Szoke T,Kayser K,Trojan I,et al.The role of microvascularization and growth/adhesion-regulatory lectins in the prognosis of non-small cell lung cancer in stage II[J].Eur J Cardiothorac Surg,2007,31(5): 783-787.
2024, Vol. 35 
