title: Quantitative proteomics identification of phosphoglycerate mutase 1 as a novel therapeutic target in hepatocellular carcinoma.
abstract: BACKGROUND
  Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide with poor prognosis due to resistance to conventional chemotherapy and limited efficacy of radiotherapy. There is an urgent need to develop novel biomarkers for early diagnosis, as well as to identify new drug targets for therapeutic interventions.
  
  
  METHODS
  54 paired HCC samples and 21 normal liver tissues were obtained from West China Hospital of Sichuan University. Informed consent was obtained from all the patients or their relatives prior to analysis, and the project was approved by the Institutional Ethics Committee of Sichuan University. Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC)-based proteomics was employed to profile the differentially expressed proteins between a HepG2 human hepatoma cell line and an immortal hepatic cell line L02. Validation of PGAM1 expression was performed by semi-quantitative RT-PCR, immunoblot and immunohistochemistry using clinical samples. shRNA expressing plasmids specifically targeting PGAM1 were designed and constructed by GenePharma Corporation (Shanghai, China), and were utilized to silence expression of PGAM1 in vitro and in vivo. Cell proliferation was measured by a combination of colony formation assay and Ki67 staining. Apoptosis was examined by flow cytometry and TUNEL assay.
  
  
  RESULTS
  A total of 63 dysregulated proteins were identified, including 51 up-regulated proteins, and 12 down-regulated proteins (over 2-fold, p < 0.01). Phosphoglycerate mutase 1 (PGAM1) was found markedly upregulated. Clinico-pathological analysis indicated that overexpression of PGAM1 was associated with 66.7% HCC, and strongly correlated with poor differentiation and decreased survival rates (p < 0.01). shRNAs-mediated repression of PGAM1 expression resulted in significant inhibition in liver cancer cell growth both in vitro and in vivo.
  
  
  CONCLUSIONS
  Our studies suggested that PGAM1 plays an important role in hepatocarcinogenesis, and should be a potential diagnostic biomarker, as well as an attractive therapeutic target for hepatocellular carcinoma.
authors.affiliation: The State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
authors.forename: Fenglian
authors.initials: F
authors.lastname: Ren
authors.forename: Hong
authors.initials: H
authors.lastname: Wu
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Lei
authors.forename: Haiyuan
authors.initials: H
authors.lastname: Zhang
authors.forename: Rui
authors.initials: R
authors.lastname: Liu
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhao
authors.forename: Xiancheng
authors.initials: X
authors.lastname: Chen
authors.forename: Dequan
authors.initials: D
authors.lastname: Zeng
authors.forename: Aiping
authors.initials: A
authors.lastname: Tong
authors.forename: Lijuan
authors.initials: L
authors.lastname: Chen
authors.forename: Yuquan
authors.initials: Y
authors.lastname: Wei
authors.forename: Canhua
authors.initials: C
authors.lastname: Huang
mesh_terms: D000328:Adult; D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002469:Cell Separation; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D009367:Neoplasm Staging; D010736:Phosphoglycerate Mutase / Q000096:biosynthesis / Q000235:genetics*; D040901:Proteomics / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023041:Xenograft Model Antitumor Assays
chemical_list: D014408:Biomarkers, Tumor; D010736:Phosphoglycerate Mutase
references.citation: J Hepatol. 2009 May;50(5):1019-28
references.pmid: 19303656
references.citation: Br J Cancer. 2009 May 5;100(9):1369-72
references.pmid: 19352381
references.citation: Gastroenterology. 2008 May;134(6):1752-63
references.pmid: 18471552
references.citation: Hepatology. 2007 Jul;46(1):200-8
references.pmid: 17596871
references.citation: J Bioenerg Biomembr. 2007 Jun;39(3):251-7
references.pmid: 17624581
references.citation: Cell. 2003 Nov 26;115(5):577-90
references.pmid: 14651849
references.citation: Oncogene. 2006 Aug 7;25(34):4633-46
references.pmid: 16892078
references.citation: Proc Natl Acad Sci U S A. 2003 May 27;100(11):6343-6
references.pmid: 12730368
references.citation: Cancer Res. 2005 Jan 1;65(1):177-85
references.pmid: 15665293
references.citation: Protein Sci. 2001 Sep;10(9):1835-46
references.pmid: 11514674
references.citation: J Biol Chem. 1988 Nov 15;263(32):16899-905
references.pmid: 2846553
references.citation: Int J Cancer. 2008 Nov 15;123(10):2377-83
references.pmid: 18729184
references.citation: Mol Cell Proteomics. 2009 Jan;8(1):70-85
references.pmid: 18723843
references.citation: Dig Surg. 2007;24(5):342-8
references.pmid: 17785977
references.citation: Nat Rev Cancer. 2004 Nov;4(11):891-9
references.pmid: 15516961
references.citation: Oncogene. 2006 Jun 26;25(27):3866-84
references.pmid: 16799628
references.citation: Proc Natl Acad Sci U S A. 2003 Nov 11;100(23):13537-42
references.pmid: 14573703
references.citation: Gene. 2001 Feb 21;264(2):273-9
references.pmid: 11250083
references.citation: Proteomics. 2008 May;8(10):2012-23
references.pmid: 18491315
references.citation: Biochim Biophys Acta. 2005 Jun 10;1740(3):403-10
references.pmid: 15949708
references.citation: Hepatology. 2008 Mar;47(3):897-907
references.pmid: 18176954
references.citation: Int J Cancer. 2001 Dec 1;94(5):662-8
references.pmid: 11745460
references.citation: FEBS Lett. 1999 Jul 23;455(3):344-8
references.pmid: 10437801
references.citation: Int J Oncol. 2008 Oct;33(4):725-31
references.pmid: 18813785
references.citation: Cancer Lett. 2009 Apr 18;276(2):125-35
references.pmid: 18790562
references.citation: Nat Biotechnol. 2005 Oct;23(10):1303-7
references.pmid: 16200062
references.citation: Electrophoresis. 2006 Apr;27(7):1417-23
references.pmid: 16568407
references.citation: J Biomed Sci. 2006 Jul;13(4):453-68
references.pmid: 16633742
references.citation: Biochem J. 1990 Jan 15;265(2):503-9
references.pmid: 2302181
references.citation: Cancer Detect Prev. 2004;28(6):443-52
references.pmid: 15582268
references.citation: Front Biosci (Landmark Ed). 2009 Jan 01;14:19-44
references.pmid: 19273052
references.citation: Liver Int. 2009 Jan;29(1):10-7
references.pmid: 19120940
references.citation: Cell. 2000 Jan 7;100(1):57-70
references.pmid: 10647931
references.citation: Gastroenterology. 2007 Jun;132(7):2557-76
references.pmid: 17570226
references.citation: J Biol Chem. 2008 Aug 15;283(33):22700-8
references.pmid: 18541534
references.citation: Mol Biol Evol. 2002 May;19(5):654-63
references.pmid: 11961099
references.citation: Expert Rev Proteomics. 2005 Aug;2(4):589-601
references.pmid: 16097891
references.citation: Curr Drug Targets. 2004 May;5(4):375-82
references.pmid: 15134220
references.citation: Carcinogenesis. 2004 Jun;25(6):865-71
references.pmid: 14764456
references.citation: J Biol Chem. 2004 Aug 20;279(34):35803-12
references.pmid: 15181008
references.citation: Cancer Immunol Immunother. 1995 Feb;40(2):73-8
references.pmid: 7882385
references.citation: Proteomics. 2006 Nov;6(22):6095-106
references.pmid: 17051635
references.citation: Br J Cancer. 2000 Jan;82(1):20-7
references.pmid: 10638961
references.citation: Horm Res. 2004;62(4):191-6
references.pmid: 15375329
references.citation: Trends Biochem Sci. 1999 Feb;24(2):68-72
references.pmid: 10098401
references.citation: Proteomics. 2006 Jan;6(2):547-58
references.pmid: 16342241
references.citation: Mol Cancer Ther. 2003 Jun;2(6):573-80
references.pmid: 12813137
references.citation: Toxicol Sci. 2008 Oct;105(2):286-94
references.pmid: 18567617
references.citation: Gastroenterology. 2004 Nov;127(5 Suppl 1):S5-S16
references.pmid: 15508102
references.citation: FEBS J. 2008 Jul;275(13):3454-69
references.pmid: 18510554
references.citation: Oncology. 2009;77(6):385-94
references.pmid: 20130421
references.citation: Cell Biol Int. 2008 Oct;32(10):1215-22
references.pmid: 18675922
references.citation: Cancer Lett. 2001 Nov 8;173(1):83-91
references.pmid: 11578813
references.citation: J Exp Med. 2000 May 15;191(10):1675-86
references.pmid: 10811861
references.citation: Science. 1956 Feb 24;123(3191):309-14
references.pmid: 13298683