Genetic alterations and oxidative metabolism in sporadic colorectal tumors from a Spanish community

Deletions of loci on chromosomes 5q, 17p, 18q, and 22q, together with the incidence of p53 mutations and amplification of the double minute‐2 gene were investigated in the sporadic colorectal tumors of 44 patients from a Spanish community. Chromosome deletions were analyzed by means of loss of heterozygosity analysis using a restriction fragment length polymorphism assay. Allelic losses were also detected by polymerase chain reaction (PCR)‐single‐stranded conformation polymorphism (SSCP) analysis of a polymorphic site in intron 2 of the p53 gene. The percentages of genetic deletions on the screened chromosomes were 39.3% (5q), 58.3% (17p), 40.9% (18q), and 40% (22q). Mutations in p53 exons 2–9 were examined by PCR‐SSCP analysis and direct sequencing of the mutated region. Twenty of 44 tumor samples (45.45%) showed mutations at various exons except for exons 2, 3, and 9, the most frequent changes being G → T transversion and C → T transition. Because oxygen‐free radicals play a role in the carcinogenesis process, we evaluated the oxidative status of the colorectal tumors. Antioxidant activities, lipid peroxidation, and DNA‐damaged product concentrations in colon tumors and normal mucosa were compared. In tumor tissues, superoxide dismutase and catalase decreased fourfold and twofold, respectively, whereas glutathione peroxidase and reduced glutathione increased threefold. Malondialdehyde and 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG) levels were twofold higher in colorectal tumors than in normal mucosa. Seven of 10 DNA tumor samples (70%) showing higher values of 8‐OHdG also had genetic alterations at different chromosomal loci. In these samples, the p53 gene was deleted or mutated in 71.4% of cases. We concluded that the observed changes in the oxidative metabolism of the tumor cells and the consecutive increase in DNA damage may potentiate the genomic instability of different chromosomal regions, leading to further cell malignancy and tumor expansion. Mol. Carcinog. 18:232–243, 1997. © 1997 Wiley‐Liss, Inc.

[1]  C. Cordon-Cardo,et al.  A new polymorphic site in intron 2 of TP53 characterizes LOH in human tumors by PCR-SSCP. , 1995, Diagnostic molecular pathology (Print).

[2]  D. Phillips,et al.  Induction of activating mutations in the human c‐Ha‐ras‐1 proto‐oncogene by oxygen free radicals , 1994, Molecular carcinogenesis.

[3]  C. Cordon-Cardo,et al.  Altered patterns of MDM2 and TP53 expression in human bladder cancer. , 1994, Journal of the National Cancer Institute.

[4]  D. S. St. Clair,et al.  Suppression of fibrosarcoma metastasis by elevated expression of manganese superoxide dismutase. , 1994, Cancer research.

[5]  P. Cerutti,et al.  Oxy-radical induced mutagenesis of hotspot codons 248 and 249 of the human p53 gene. , 1994, Oncogene.

[6]  J. Willson,et al.  Sensitive enzymatic cycling assay for glutathione: measurements of glutathione content and its modulation by buthionine sulfoximine in vivo and in vitro in human colon cancer. , 1994, Cancer research.

[7]  Z. F. Liu,et al.  Allelic loss of chromosome 18q and prognosis in colorectal cancer. , 1994, The New England journal of medicine.

[8]  C. Harris,et al.  Geographic variation of p53 mutational profile in nonmalignant human liver. , 1994, Science.

[9]  A. Fornace,et al.  The p53-dependent γ-Ray Response of GADD45 , 1994 .

[10]  M. Fukayama,et al.  Molecular genetics for clinical management of colorectal carcinoma. 17p, 18q, and 22q loss of heterozygosity and decreased DCC expression are correlated with the metastatic potential , 1994, Cancer.

[11]  C. Cordon-Cardo,et al.  p53 mutations in human bladder cancer: Genotypic versus phenotypic patterns , 1994, International journal of cancer.

[12]  A. Levine,et al.  Molecular abnormalities of mdm2 and p53 genes in adult soft tissue sarcomas. , 1994, Cancer research.

[13]  R. Mangues,et al.  Absence of MDM‐2 gene amplification in experimentally induced tumors regardless of p53 status , 1994, Molecular carcinogenesis.

[14]  H. Kovar,et al.  Narrow spectrum of infrequent p53 mutations and absence of MDM2 amplification in Ewing tumours. , 1993, Oncogene.

[15]  T. Sellers,et al.  Reduced risk of colon cancer with high intake of vitamin E: the Iowa Women's Health Study. , 1993, Cancer research.

[16]  A. Levine,et al.  Mapping of the p53 and mdm-2 interaction domains. , 1993, Molecular and cellular biology.

[17]  S. Hamilton,et al.  The molecular genetics of colorectal neoplasia. , 1993, Gastroenterology.

[18]  P. Meltzer,et al.  p53 Mutation and MDM2 amplification in human soft tissue sarcomas. , 1993, Cancer research.

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

[20]  J. Marx.  New colon cancer gene discovered. , 1993, Science.

[21]  C. Boland,et al.  Genetics, natural history, tumor spectrum, and pathology of hereditary nonpolyposis colorectal cancer: an updated review. , 1993, Gastroenterology.

[22]  R. Lothe,et al.  The TP53 tumour suppressor gene in colorectal carcinomas. I. Genetic alterations on chromosome 17. , 1993, British Journal of Cancer.

[23]  B. Vogelstein,et al.  A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia , 1992, Cell.

[24]  M. Kastan,et al.  Wild-type p53 is a cell cycle checkpoint determinant following irradiation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[25]  A. Levine,et al.  Two distinct mechanisms alter p53 in breast cancer: mutation and nuclear exclusion. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[26]  P. Meltzer,et al.  Amplification of a gene encoding a p53-associated protein in human sarcomas , 1992, Nature.

[27]  A. Levine,et al.  The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation , 1992, Cell.

[28]  K. Frenkel,et al.  Suppression of tumor promoter-induced oxidative events and DNA damage in vivo by sarcophytol A: a possible mechanism of antipromotion. , 1992, Cancer research.

[29]  B. Vogelstein,et al.  Participation of p53 protein in the cellular response to DNA damage. , 1991, Cancer research.

[30]  R. Metcalf,et al.  Genetic analysis of human esophageal tumors from two high incidence geographic areas: frequent p53 base substitutions and absence of ras mutations. , 1991, Cancer research.

[31]  J. Solomon,et al.  Quantitative high-performance liquid chromatography analysis of DNA oxidized in vitro and in vivo. , 1991, Analytical biochemistry.

[32]  A. Levine,et al.  The p53 tumour suppressor gene , 1991, Nature.

[33]  D. George,et al.  Tumorigenic potential associated with enhanced expression of a gene that is amplified in a mouse tumor cell line. , 1991, The EMBO journal.

[34]  J. Wands,et al.  Selective G to T mutations of p53 gene in hepatocellular carcinoma from southern Africa , 1991, Nature.

[35]  A. Grollman,et al.  Insertion of specific bases during DNA synthesis past the oxidation-damaged base 8-oxodG , 1991, Nature.

[36]  B. Vogelstein,et al.  p53 gene mutations occur in combination with 17p allelic deletions as late events in colorectal tumorigenesis. , 1990, Cancer research.

[37]  J. Bufill,et al.  Colorectal cancer: evidence for distinct genetic categories based on proximal or distal tumor location. , 1990, Annals of internal medicine.

[38]  R. Floyd.  The role of 8-hydroxyguanine in carcinogenesis. , 1990, Carcinogenesis.

[39]  J. Essigmann,et al.  Mechanistic studies of ionizing radiation and oxidative mutagenesis: genetic effects of a single 8-hydroxyguanine (7-hydro-8-oxoguanine) residue inserted at a unique site in a viral genome. , 1990, Biochemistry.

[40]  B. Vogelstein,et al.  A genetic model for colorectal tumorigenesis , 1990, Cell.

[41]  E. Stanbridge.  Identifying tumor suppressor genes in human colorectal cancer. , 1990, Science.

[42]  F. Collins,et al.  Mutations in the p53 gene occur in diverse human tumour types , 1989, Nature.

[43]  P. Okunieff,et al.  Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. , 1989, Cancer research.

[44]  T. Sekiya,et al.  Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. , 1989, Genomics.

[45]  R. Weinberg,et al.  Oncogenes, antioncogenes, and the molecular bases of multistep carcinogenesis. , 1989, Cancer research.

[46]  A. Levine,et al.  The p53 proto-oncogene can act as a suppressor of transformation , 1989, Cell.

[47]  M. Leppert,et al.  Allelic Loss in Colorectal Carcinoma , 1989 .

[48]  Y. Nakamura,et al.  Clinical and pathological associations with allelic loss in colorectal carcinoma [corrected]. , 1989, JAMA.

[49]  Y. Nakamura,et al.  Allelotype of colorectal carcinomas. , 1989, Science.

[50]  Y. Nakamura,et al.  Genetic alterations during colorectal-tumor development. , 1988, The New England journal of medicine.

[51]  B. Ames,et al.  Normal oxidative damage to mitochondrial and nuclear DNA is extensive. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[52]  R. Floyd,et al.  Hydroxyl free radical mediated formation of 8-hydroxyguanine in isolated DNA. , 1988, Archives of biochemistry and biophysics.

[53]  K. Sugio,et al.  Loss of constitutional heterozygosity in colon carcinoma from patients with familial polyposis coli , 1988, Nature.

[54]  B. Ames,et al.  Oxygen radicals and human disease. , 1987, Annals of internal medicine.

[55]  H. Kasai,et al.  Misreading of DNA templates containing 8-hydroxydeoxyguanosine at the modified base and at adjacent residues , 1987, Nature.

[56]  B. Halliwell,et al.  Free radicals in biology and medicine , 1985 .

[57]  P. Cerutti.  Prooxidant states and tumor promotion. , 1985, Science.

[58]  H. Sies,et al.  Identification and quantitation of glutathione in hepatic protein mixed disulfides and its relationship to glutathione disulfide. , 1983, Biochemical pharmacology.

[59]  M. Uchiyama,et al.  Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. , 1978, Analytical biochemistry.

[60]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[61]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[62]  G. Thomas,et al.  Association of p53 mutations with short survival in colorectal cancer. , 1994, Gastroenterology.

[63]  K. Guyton,et al.  Oxidative mechanisms in carcinogenesis. , 1993, British medical bulletin.

[64]  J. Gutteridge,et al.  Free radicals in disease processes: a compilation of cause and consequence. , 1993, Free radical research communications.

[65]  K. Frenkel.  Carcinogen-mediated oxidant formation and oxidative DNA damage. , 1992, Pharmacology & therapeutics.

[66]  A. Fornace.  Mammalian genes induced by radiation; activation of genes associated with growth control. , 1992, Annual review of genetics.

[67]  Y. Sun,et al.  Free radicals, antioxidant enzymes, and carcinogenesis. , 1990, Free radical biology & medicine.