Microsatellite instability associated with hepatocarcinogenesis.

BACKGROUND/AIMS The biological and clinicopathological significance of microsatellite instability in hepatocellular carcinoma still remains to be determined. The aim of this study was to assess the role of microsatellite instability in hepatocarcinogenesis. METHODS Genomic DNA extracted from 38 fresh samples of hepatocellular carcinoma was amplified by polymerase chain reaction using 29 fluorescence-labeled microsatellite markers and analyzed using a semi-automated laser scanning system. Associations between the incidence of replication error and the clinicopathological features of hepatocellular carcinoma were evaluated. Since reference DNA was extracted from corresponding fresh samples of non-cancerous liver tissue, the incidence of microsatellite instability in non-cancerous liver tissues was not assessed in this study. RESULTS Four (11%) hepatocellular carcinomas had a replication error in one or two microsatellite markers; they were all poorly differentiated hepatocellular carcinomas. The incidence of replication error correlated significantly with the histological differentiation of the tumor (p<0.05) and with portal vein involvement (p<0.05). All four hepatocellular carcinomas with replication errors showed loss of heterozygosity in one or more of the 29 markers we examined. No replication errors were detected in six markers in the coding regions of the BAX, insulin-like growth factor II receptor, transforming growth factor-beta type II receptor, E2F-4, hMSH3 and hMSH6 genes. CONCLUSIONS The results of this study indicate that: (1) microsatellite instability is a rare event during hepatocarcinogenesis and may be specifically associated with progression of hepatocellular carcinoma; and (2) frame-shift mutation in the above six genes is not a common mechanism involved in progression of this cancer.

[1]  Cécile Fizames,et al.  The 1993–94 Généthon human genetic linkage map , 1994, Nature Genetics.

[2]  H. Yamamoto,et al.  Somatic frameshift mutations in DNA mismatch repair and proapoptosis genes in hereditary nonpolyposis colorectal cancer. , 1998, Cancer research.

[3]  R. Weinberg,et al.  E2F-4 and E2F-5, two members of the E2F family, are expressed in the early phases of the cell cycle. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[4]  K. Kinzler,et al.  Inactivation of the type II TGF-beta receptor in colon cancer cells with microsatellite instability. , 1995, Science.

[5]  E. Wilson,et al.  Reduced androgen receptor gene expression with first exon CAG repeat expansion. , 1996, Molecular endocrinology.

[6]  Noriyuki Moriyama,et al.  Pathology of small hepatocellular carcinoma. A proposal for a new gross classification , 1987, Cancer.

[7]  G. Klintmalm,et al.  Microsatellite instability in human hepatocellular carcinoma: relationship to p53 abnormalities. , 2008, Liver.

[8]  J. Bruix,et al.  Androgen receptors in hepatocellular carcinoma and surrounding liver: relationship with tumor size and recurrence rate after surgical resection. , 1995, Journal of hepatology.

[9]  Y. Nakamura,et al.  Detection of loss of heterozygosity at the human TP53 locus using a dinucleotide repeat polymorphism , 1992, Genes, chromosomes & cancer.

[10]  K. M. Mulder,et al.  Differential sensitivity of subclasses of human colon carcinoma cell lines to the growth inhibitory effects of transforming growth factor-beta 1. , 1989, Experimental cell research.

[11]  J. Mattick,et al.  'Touchdown' PCR to circumvent spurious priming during gene amplification. , 1991, Nucleic acids research.

[12]  K. Kinzler,et al.  Lessons from Hereditary Colorectal Cancer , 1996, Cell.

[13]  K. Kinzler,et al.  Clues to the pathogenesis of familial colorectal cancer. , 1993, Science.

[14]  K. Kinzler,et al.  Analysis of mismatch repair genes in hereditary non–polyposis colorectal cancer patients , 1996, Nature Medicine.

[15]  M. Leppert,et al.  A CA repeat 30-70 KB downstream from the adenomatous polyposis coli (APC) gene. , 1991, Nucleic acids research.

[16]  A. Kurt,et al.  Microsatellite alterations in plasma DNA of small cell lung cancer patients , 1996, Nature Medicine.

[17]  S N Thibodeau,et al.  Microsatellite instability in cancer of the proximal colon. , 1993, Science.

[18]  S. Hirohashi,et al.  The E‐cadherin gene is silenced by CpG methylation in human hepatocellular carcinomas , 1997, International journal of cancer.

[19]  Y. Nakamura,et al.  Genetic instability in pancreatic cancer and poorly differentiated type of gastric cancer. , 1993, Cancer research.

[20]  H. Yamamoto,et al.  Frameshift somatic mutations in gastrointestinal cancer of the microsatellite mutator phenotype. , 1997, Cancer research.

[21]  D. S. Rath,et al.  Tetranucleotide repeat polymorphism at the human beta-actin related pseudogene H-beta-Ac-psi-2 (ACTBP2). , 1992, Nucleic acids research.

[22]  Y S Erozan,et al.  Molecular Detection of Primary Bladder Cancer by Microsatellite Analysis , 1996, Science.

[23]  S. Hirohashi,et al.  Increased DNA Methyltransferase Expression Is Associated with an Early Stage of Human Hepatocarcinogenesis , 1997, Japanese journal of cancer research : Gann.

[24]  M. Loda,et al.  Methylation of the hMLH1 promoter correlates with lack of expression of hMLH1 in sporadic colon tumors and mismatch repair-defective human tumor cell lines. , 1997, Cancer research.

[25]  R. Weinberg,et al.  Expression cloning of the TGF-β type II receptor, a functional transmembrane serine/threonine kinase , 1992, Cell.

[26]  Y S Erozan,et al.  Microsatellite alterations as clonal markers for the detection of human cancer. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[27]  S. Hirohashi,et al.  Allele loss on chromosome 16 associated with progression of human hepatocellular carcinoma. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[28]  B. Leggett,et al.  Microsatellite instability in the insulin–like growth factor II receptor gene in gastrointestinal tumours , 1996, Nature Genetics.

[29]  S. Kern,et al.  Analysis of replication error (RER+) phenotypes in cervical carcinoma. , 1996, Cancer research.

[30]  H. Bismuth,et al.  Frequent microsatellite instability in post hepatitis B viral cirrhosis. , 1996, Oncogene.

[31]  A. Chapelle,et al.  Loss of the wild type MLH1 gene is a feature of hereditary nonpolyposis colorectal cancer , 1994, Nature Genetics.

[32]  S. Hirohashi,et al.  Aberrant DNA Methylation on Chromosome 16 Is an Early Event in Hepatocarcinogenesis , 1996, Japanese journal of cancer research : Gann.

[33]  J. Rüschoff,et al.  Diagnostic microsatellite instability: definition and correlation with mismatch repair protein expression. , 1997, Cancer research.

[34]  D. Kwiatkowski,et al.  Dinucleotide repeat polymorphism at the IFNA locus (9p22). , 1992, Human molecular genetics.

[35]  G. Gyapay,et al.  A second-generation linkage map of the human genome , 1992, Nature.

[36]  K. Kinzler,et al.  Genetic instability in colorectal cancers , 1997, Nature.

[37]  Darryl Shibata,et al.  Ubiquitous somatic mutations in simple repeated sequences reveal a new mechanism for colonic carcinogenesis , 1993, Nature.

[38]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[39]  L. Aaltonen,et al.  Semiautomated assessment of loss of heterozygosity and replication error in tumors. , 1996, Cancer research.

[40]  A. Knudson,et al.  Genetics of human cancer. , 1986, Annual review of genetics.

[41]  L. Peltonen,et al.  An Alu variable polyA repeat polymorphism upstream of L-myc at 1p32. , 1992, Human molecular genetics.

[42]  J. Weber,et al.  Dinucleotide repeat polymorphism at the D15S87 locus. , 1990, Nucleic acids research.

[43]  Hiroyuki Yamamoto,et al.  Frameshift mutator mutations , 1996, Nature.

[44]  J C Reed,et al.  Somatic Frameshift Mutations in the BAX Gene in Colon Cancers of the Microsatellite Mutator Phenotype , 1997, Science.

[45]  D. Rifkin,et al.  Cellular activation of latent transforming growth factor beta requires binding to the cation-independent mannose 6-phosphate/insulin-like growth factor type II receptor. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[46]  N. Matsubara,et al.  Mutations of E2F-4 trinucleotide repeats in colorectal cancer with microsatellite instability. , 1996, Biochemical and biophysical research communications.

[47]  S. Korsmeyer,et al.  Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death , 1993, Cell.

[48]  David Sidransky,et al.  Microsatellite alterations in serum DNA of head and neck cancer patients , 1996, Nature Medicine.

[49]  Sajeev P. Cherian,et al.  Microsatellite instability and loss of heterozygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis , 1998, Hepatology.

[50]  J. Fujimoto,et al.  Flow cytometric dna analysis of hepatocellular carcinoma , 1991, Cancer.

[51]  K. Kinzler,et al.  Mutations of GTBP in genetically unstable cells. , 1995, Science.

[52]  S. Hirohashi,et al.  Early stages of multistep hepatocarcinogenesis: adenomatous hyperplasia and early hepatocellular carcinoma. , 1991, Human pathology.