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Proc Natl Acad Sci U S A. Apr 30, 1996; 93(9): 4045–4050.

Increased cytosine DNA-methyltransferase activity is target-cell-specific and an early event in lung cancer.


The association between increased DNA-methyltransferase (DNA-MTase) activity and tumor development suggest a fundamental role for this enzyme in the initiation and progression of cancer. A true functional role for DNA-MTase in the neoplastic process would be further substantiated if the target cells affected by the initiating carcinogen exhibit changes in enzyme activity. This hypothesis was addressed by examining DNA-MTase activity in alveolar type II (target) and Clara (nontarget) cells from A/J and C3H mice that exhibit high and low susceptibility, respectively, for lung tumor formation. Increased DNA-MTase activity was found only in the target alveolar type II cells of the susceptible A/J mouse and caused a marked increase in overall DNA methylation in these cells. Both DNA-MTase and DNA methylation changes were detected 7 days after carcinogen exposure and, thus, were early events in neoplastic evolution. Increased gene expression was also detected by RNA in situ hybridization in hypertrophic alveolar type II cells of carcinogen-treated A/J mice, indicating that elevated levels of expression may be a biomarker for premalignancy. Enzyme activity increased incrementally during lung cancer progression and coincided with increased expression of the DNA-MTase activity are strongly associated with neoplastic development and constitute a key step in carcinogenesis. The detection of premalignant lung disease through increased DNA-MTase expression and the possibility of blocking the deleterious effects of this change with specific inhibitors will offer new intervention strategies for lung cancer.

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  • Bestor T, Laudano A, Mattaliano R, Ingram V. Cloning and sequencing of a cDNA encoding DNA methyltransferase of mouse cells. The carboxyl-terminal domain of the mammalian enzymes is related to bacterial restriction methyltransferases. J Mol Biol. 1988 Oct 20;203(4):971–983. [PubMed]
  • Issa JP, Vertino PM, Wu J, Sazawal S, Celano P, Nelkin BD, Hamilton SR, Baylin SB. Increased cytosine DNA-methyltransferase activity during colon cancer progression. J Natl Cancer Inst. 1993 Aug 4;85(15):1235–1240. [PubMed]
  • Wu J, Issa JP, Herman J, Bassett DE, Jr, Nelkin BD, Baylin SB. Expression of an exogenous eukaryotic DNA methyltransferase gene induces transformation of NIH 3T3 cells. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8891–8895. [PMC free article] [PubMed]
  • Laird PW, Jackson-Grusby L, Fazeli A, Dickinson SL, Jung WE, Li E, Weinberg RA, Jaenisch R. Suppression of intestinal neoplasia by DNA hypomethylation. Cell. 1995 Apr 21;81(2):197–205. [PubMed]
  • Herman JG, Latif F, Weng Y, Lerman MI, Zbar B, Liu S, Samid D, Duan DS, Gnarra JR, Linehan WM, et al. Silencing of the VHL tumor-suppressor gene by DNA methylation in renal carcinoma. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9700–9704. [PMC free article] [PubMed]
  • Ottaviano YL, Issa JP, Parl FF, Smith HS, Baylin SB, Davidson NE. Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells. Cancer Res. 1994 May 15;54(10):2552–2555. [PubMed]
  • Issa JP, Ottaviano YL, Celano P, Hamilton SR, Davidson NE, Baylin SB. Methylation of the oestrogen receptor CpG island links ageing and neoplasia in human colon. Nat Genet. 1994 Aug;7(4):536–540. [PubMed]
  • Steenman MJ, Rainier S, Dobry CJ, Grundy P, Horon IL, Feinberg AP. Loss of imprinting of IGF2 is linked to reduced expression and abnormal methylation of H19 in Wilms' tumour. Nat Genet. 1994 Jul;7(3):433–439. [PubMed]
  • Merlo A, Herman JG, Mao L, Lee DJ, Gabrielson E, Burger PC, Baylin SB, Sidransky D. 5' CpG island methylation is associated with transcriptional silencing of the tumour suppressor p16/CDKN2/MTS1 in human cancers. Nat Med. 1995 Jul;1(7):686–692. [PubMed]
  • Wales MM, Biel MA, el Deiry W, Nelkin BD, Issa JP, Cavenee WK, Kuerbitz SJ, Baylin SB. p53 activates expression of HIC-1, a new candidate tumour suppressor gene on 17p13.3. Nat Med. 1995 Jun;1(6):570–577. [PubMed]
  • Barker D, Schafer M, White R. Restriction sites containing CpG show a higher frequency of polymorphism in human DNA. Cell. 1984 Jan;36(1):131–138. [PubMed]
  • Rideout WM, 3rd, Coetzee GA, Olumi AF, Jones PA. 5-Methylcytosine as an endogenous mutagen in the human LDL receptor and p53 genes. Science. 1990 Sep 14;249(4974):1288–1290. [PubMed]
  • Belinsky SA, Devereux TR, Foley JF, Maronpot RR, Anderson MW. Role of the alveolar type II cell in the development and progression of pulmonary tumors induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in the A/J mouse. Cancer Res. 1992 Jun 1;52(11):3164–3173. [PubMed]
  • Malkinson AM. The genetic basis of susceptibility to lung tumors in mice. Toxicology. 1989 Mar;54(3):241–271. [PubMed]
  • Belinsky SA, Lechner JF, Johnson NF. An improved method for the isolation of type II and Clara cells from mice. In Vitro Cell Dev Biol Anim. 1995 May;31(5):361–366. [PubMed]
  • Foley JF, Anderson MW, Stoner GD, Gaul BW, Hardisty JF, Maronpot RR. Proliferative lesions of the mouse lung: progression studies in strain A mice. Exp Lung Res. 1991 Mar-Apr;17(2):157–168. [PubMed]
  • Adams RL, Rinaldi A, Seivwright C. Microassay for DNA methyltransferase. J Biochem Biophys Methods. 1991 Jan;22(1):19–22. [PubMed]
  • Iezzoni JC, Kang JH, Bucana CD, Reed JA, Brigati DJ. Rapid colorimetric detection of epidermal growth factor receptor mRNA by in situ hybridization. J Clin Lab Anal. 1993;7(5):247–251. [PubMed]
  • Radinsky R, Bucana CD, Ellis LM, Sanchez R, Cleary KR, Brigati DJ, Fidler IJ. A rapid colorimetric in situ messenger RNA hybridization technique for analysis of epidermal growth factor receptor in paraffin-embedded surgical specimens of human colon carcinomas. Cancer Res. 1993 Mar 1;53(5):937–943. [PubMed]
  • Devereux TR, Anderson MW, Belinsky SA. Role of ras protooncogene activation in the formation of spontaneous and nitrosamine-induced lung tumors in the resistant C3H mouse. Carcinogenesis. 1991 Feb;12(2):299–303. [PubMed]
  • Ryan J, Barker PE, Nesbitt MN, Ruddle FH. KRAS2 as a genetic marker for lung tumor susceptibility in inbred mice. J Natl Cancer Inst. 1987 Dec;79(6):1351–1357. [PubMed]
  • You M, Wang Y, Stoner G, You L, Maronpot R, Reynolds SH, Anderson M. Parental bias of Ki-ras oncogenes detected in lung tumors from mouse hybrids. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5804–5808. [PMC free article] [PubMed]
  • Chen B, Johanson L, Wiest JS, Anderson MW, You M. The second intron of the K-ras gene contains regulatory elements associated with mouse lung tumor susceptibility. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1589–1593. [PMC free article] [PubMed]
  • Marx J. Forging a path to the nucleus. Science. 1993 Jun 11;260(5114):1588–1590. [PubMed]
  • Rouleau J, Tanigawa G, Szyf M. The mouse DNA methyltransferase 5'-region. A unique housekeeping gene promoter. J Biol Chem. 1992 Apr 15;267(11):7368–7377. [PubMed]
  • Rouleau J, MacLeod AR, Szyf M. Regulation of the DNA methyltransferase by the Ras-AP-1 signaling pathway. J Biol Chem. 1995 Jan 27;270(4):1595–1601. [PubMed]
  • MacLeod AR, Rouleau J, Szyf M. Regulation of DNA methylation by the Ras signaling pathway. J Biol Chem. 1995 May 12;270(19):11327–11337. [PubMed]
  • MacLeod AR, Szyf M. Expression of antisense to DNA methyltransferase mRNA induces DNA demethylation and inhibits tumorigenesis. J Biol Chem. 1995 Apr 7;270(14):8037–8043. [PubMed]
  • Hammond WG, Yellin A, Gabriel A, Paladugu RR, Azumi N, Hill LR, Benfield JR. Effects of 5-azacytidine in Syrian golden hamsters: toxicity, tumorigenicity, and differential modulation of bronchial carcinogenesis. Exp Mol Pathol. 1990 Aug;53(1):34–51. [PubMed]
  • Antequera F, Boyes J, Bird A. High levels of de novo methylation and altered chromatin structure at CpG islands in cell lines. Cell. 1990 Aug 10;62(3):503–514. [PubMed]
  • de Bustros A, Nelkin BD, Silverman A, Ehrlich G, Poiesz B, Baylin SB. The short arm of chromosome 11 is a "hot spot" for hypermethylation in human neoplasia. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5693–5697. [PMC free article] [PubMed]
  • Schmutte C, Yang AS, Beart RW, Jones PA. Base excision repair of U:G mismatches at a mutational hotspot in the p53 gene is more efficient than base excision repair of T:G mismatches in extracts of human colon tumors. Cancer Res. 1995 Sep 1;55(17):3742–3746. [PubMed]
  • Sozzi G, Miozzo M, Donghi R, Pilotti S, Cariani CT, Pastorino U, Della Porta G, Pierotti MA. Deletions of 17p and p53 mutations in preneoplastic lesions of the lung. Cancer Res. 1992 Nov 1;52(21):6079–6082. [PubMed]
  • Pastorino U, Sozzi G, Miozzo M, Tagliabue E, Pilotti S, Pierotti MA. Genetic changes in lung cancer. J Cell Biochem Suppl. 1993;17F:237–248. [PubMed]
  • Richel DJ, Colly LP, Kluin-Nelemans JC, Willemze R. The antileukaemic activity of 5-Aza-2 deoxycytidine (Aza-dC) in patients with relapsed and resistant leukaemia. Br J Cancer. 1991 Jul;64(1):144–148. [PMC free article] [PubMed]
  • Pinto A, Zagonel V. 5-Aza-2'-deoxycytidine (Decitabine) and 5-azacytidine in the treatment of acute myeloid leukemias and myelodysplastic syndromes: past, present and future trends. Leukemia. 1993 May;7 (Suppl 1):51–60. [PubMed]

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