title: Methylation-silencing RCC1 expression is associated with tumorigenesis and depth of invasion in gastric cancer.
abstract: BACKGROUND
  Regulator of chromosome condensation 1 (RCC1) is a critical cell cycle regulator. We firstly identified RCC1 gene hypermethylation in gastric tumor tissues using the differential methylation hybridization (DMH) microarray, but the role of RCC1 in the pathogenesis of gastric carcinoma is largely unknown.
  
  
  METHODS
  Three gastric cancer cell lines (AGS, MKN45, and TSGH9201) were used to analyze RCC1 gene methylation, mRNA and protein expressions. Furthermore, 85 pairs of matched human gastric carcinoma samples in a tissue microarray were used to analyze RCC1 expression by immunohistochemistry staining.
  
  
  RESULTS
  A differential methylation pattern was found in TSGH9201 (100%), MKN45 (87%), and AGS (62%) cell lines at the 9th CpG site of RCC1 exon 1. RCC1 mRNA and protein expressions in AGS cells were significantly higher than in TSGH9201 and MKN45 cell lines (P < 0.05). Tissue array data showed that RCC1 expression was detected in 21% (18/85) of gastric carcinoma tissues and in 80% (76/95) of adjacent non-tumor tissues. The expression of RCC1 in gastric carcinoma tissues was significantly lower than in adjacent non-tumor tissues (P < 0.001). Furthermore, an association between RCC1 expression and clinicopathological features showed that RCC1 expression was closely correlated with tumor differentiation and depth of invasion (P < 0.05).
  
  
  CONCLUSIONS
  Our data indicate that RCC1 expression is frequently lost in poorly differentiated gastric cell lines and gastric carcinoma tissues. Loss of RCC1 expression is correlated with tumor differentiation and depth of invasion. These findings suggest that RCC1 may play a tumor suppressor role in gastric carcinoma.
authors.affiliation: Department of Life Sciences, and Agricultural Biotechnology Center, National Chung Hsing UniversityTaichung 402, Taiwan; Department of General Surgery, Taichung Veterans General HospitalTaichung 407, Taiwan.
authors.forename: Yi-Ling
authors.initials: YL
authors.lastname: Lin
authors.affiliation: Department of Bioresources, Da-Yeh University Changhwa 515, Taiwan.
authors.forename: Hsiao-Ling
authors.initials: HL
authors.lastname: Chen
authors.affiliation: Department of General Surgery, Taichung Veterans General HospitalTaichung 407, Taiwan; School of Medicine, Chung-Shan Medical UniversityTaichung 402, Taiwan.
authors.forename: Shao-Bin
authors.initials: SB
authors.lastname: Cheng
authors.affiliation: Department of General Surgery, Taichung Veterans General Hospital Taichung 407, Taiwan.
authors.forename: Dah-Cherng
authors.initials: DC
authors.lastname: Yeh
authors.affiliation: Department of General Surgery, Taichung Veterans General Hospital Taichung 407, Taiwan.
authors.forename: Chu-Chun
authors.initials: CC
authors.lastname: Huang
authors.affiliation: Department of Surgery, Faculty of Medicine, National Yang-Ming University Taipei 112, Taiwan.
authors.forename: Fang-Ku
authors.initials: FK
authors.lastname: P'eng
authors.affiliation: Department of Life Sciences, and Agricultural Biotechnology Center, National Chung Hsing University Taichung 402, Taiwan.
authors.forename: Tung-Chou
authors.initials: TC
authors.lastname: Tsai
authors.affiliation: Department of General Surgery, Taichung Veterans General HospitalTaichung 407, Taiwan; Department of Surgery, Faculty of Medicine, National Yang-Ming UniversityTaipei 112, Taiwan; Department of Surgery, Chung-Shan Medical UniversityTaichung 402, Taiwan; Department of Surgery, Taipei Medical UniversityTaipei 110, Taiwan.
authors.forename: Cheng-Chung
authors.initials: CC
authors.lastname: Wu
authors.affiliation: Department of Life Sciences, and Agricultural Biotechnology Center, National Chung Hsing UniversityTaichung 402, Taiwan; Rong-Hsing Translational Medicine Center, and iEGG Center, National Chung Hsing UniversityTaichung 402, Taiwan.
authors.forename: Chuan-Mu
authors.initials: CM
authors.lastname: Chen
mesh_terms: D001483:Base Sequence; D063646:Carcinogenesis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002277:Carcinoma / Q000235:genetics / Q000378:metabolism* / Q000556:secondary; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002465:Cell Movement; D018899:CpG Islands; D019175:DNA Methylation; D015536:Down-Regulation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D060787:Neoplasm Grading; D009361:Neoplasm Invasiveness; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D046888:Tissue Array Analysis; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D020662:Guanine Nucleotide Exchange Factors; D009687:Nuclear Proteins; C062521:RCC1 protein, human; D012333:RNA, Messenger; D025521:Tumor Suppressor Proteins
keywords: DMH microarray; DNA methylation; RCC1; gastric carcinoma; immunohistochemistry; invasion; tumor tissue array
references.citation: Am J Pathol. 2000 Sep;157(3):787-94
references.pmid: 10980118
references.citation: Dev Biol. 2004 Jun 1;270(1):106-21
references.pmid: 15136144
references.citation: PLoS One. 2012;7(11):e49683
references.pmid: 23152928
references.citation: PLoS One. 2014;9(3):e90818
references.pmid: 24595062
references.citation: Genomics. 1994 Oct;23(3):719-21
references.pmid: 7851910
references.citation: PLoS One. 2015;10(2):e0115968
references.pmid: 25700270
references.citation: EMBO J. 1991 Jun;10(6):1555-64
references.pmid: 1851087
references.citation: Oncotarget. 2015 Apr 30;6(12):10222-38
references.pmid: 25912305
references.citation: Cancer Res. 2001 Dec 1;61(23):8375-80
references.pmid: 11731411
references.citation: Lancet. 2009 Aug 8;374(9688):477-90
references.pmid: 19625077
references.citation: Vaccine. 2008 Jun 2;26(23):2882-9
references.pmid: 18450335
references.citation: Genes Dev. 2001 Nov 15;15(22):2945-9
references.pmid: 11711430
references.citation: Int J Cancer. 2011 Dec 1;129(11):2600-10
references.pmid: 21960261
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Cancer. 1988 Oct 15;62(8 Suppl):1676-9
references.pmid: 3167785
references.citation: Acta Pathol Microbiol Scand. 1965;64:31-49
references.pmid: 14320675
references.citation: Am J Pathol. 2003 Jul;163(1):37-45
references.pmid: 12819009
references.citation: World J Gastroenterol. 2015 Apr 28;21(16):4954-60
references.pmid: 25945009
references.citation: Int J Surg Oncol. 2012;2012:868645
references.pmid: 22778942
references.citation: Int J Cancer. 2009 Aug 1;125(3):666-73
references.pmid: 19382179
references.citation: Genes Dev. 1987 Aug;1(6):585-93
references.pmid: 3678831
references.citation: Nature. 1991 Nov 7;354(6348):80-2
references.pmid: 1944575
references.citation: Aliment Pharmacol Ther. 2002 Jul;16(7):1209-16
references.pmid: 12144569
references.citation: Phytomedicine. 2014 Oct 15;21(12):1708-16
references.pmid: 25442281
references.citation: J Pathol. 2004 Jun;203(2):681-7
references.pmid: 15141383
references.citation: Gan. 1968 Jun;59(3):251-8
references.pmid: 5726267
references.citation: Histopathology. 2005 May;46(5):505-14
references.pmid: 15842632
references.citation: Cancer Res. 1992 Dec 15;52(24):6735-40
references.pmid: 1458460
references.citation: Cancer Res. 1994 Apr 1;54(7 Suppl):1941s-1943s
references.pmid: 8137316
references.citation: Hum Mol Genet. 1992 Nov;1(8):559-63
references.pmid: 1338691
references.citation: Carcinogenesis. 2011 Sep;32(9):1305-14
references.pmid: 21803736
references.citation: Clin Cancer Res. 2003 Sep 15;9(11):4034-42
references.pmid: 14519624
references.citation: Lab Invest. 1998 Sep;78(9):1143-53
references.pmid: 9759658
references.citation: J Surg Oncol. 1995 Jan;58(1):17-24
references.pmid: 7823569
references.citation: Cancer. 2000 Apr 15;88(8):1801-6
references.pmid: 10760755
references.citation: Oncotarget. 2014 Dec 15;5(23):12273-90
references.pmid: 25401518
references.citation: Int J Cancer. 2010 Dec 15;127(12):2893-917
references.pmid: 21351269
references.citation: Cancer Res. 2002 Jun 1;62(11):3214-20
references.pmid: 12036936
references.citation: J Nutr. 2002 Aug;132(8 Suppl):2430S-2434S
references.pmid: 12163706
references.citation: J Urol. 2014 Nov;192(5):1528-33
references.pmid: 24704013
references.citation: Carcinogenesis. 2015 May;36(5):553-63
references.pmid: 25784376
references.citation: Methods Mol Biol. 2009;472:467-77
references.pmid: 19107449
references.citation: Lancet Oncol. 2008 Mar;9(3):279-87
references.pmid: 18308253
references.citation: World J Gastroenterol. 2013 Feb 28;19(8):1182-92
references.pmid: 23482412
references.citation: Cell. 1978 Oct;15(2):475-83
references.pmid: 719750
references.citation: Gene. 2011 Dec 15;490(1-2):6-14
references.pmid: 21963386
references.citation: Cancer Res. 1994 Sep 1;54(17):4750-5
references.pmid: 8062274
references.citation: Nature. 1993 Oct 14;365(6447):661-3
references.pmid: 8413630
references.citation: Biomed Res Int. 2014;2014:378358
references.pmid: 25197641