Induction of Alpha-fetoprotein Synthesis in Differentiating F9 Teratocarcinoma Cells is Accompanied by a Genome-wide Loss of DNA Methylation

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1984 May 1

PMID 6203029

Citations 25

Authors

P R Young

S M Tilghman

Affiliations

Soon will be listed here.

Abstract

F9 teratocarcinoma cells can be grown as monolayers or aggregates, and upon treatment with retinoic acid they will differentiate into parietal or visceral endoderm, respectively. Visceral endoderm specifically synthesizes alpha-fetoprotein and albumin mRNAs, which are not found in parietal endoderm. In contrast, both endoderms produce enhanced levels of the major histocompatibility antigen (H2) mRNA compared with F9 cells. F9 cells contain highly methylated DNA as judged by restriction enzyme digestion. However, upon differentiation into visceral endoderm, there is a genome-wide loss of methylation in induced, silent, and constitutively expressed genes. Experiments in which methylation loss is induced via the methyltransferase inhibitor 5-azacytidine result in no induction of alpha-fetoprotein mRNA and no morphological differentiation, suggesting that methylation loss alone is not sufficient to induce the visceral endoderm phenotype. Likewise, 5-azacytidine treatment of differentiated cells does not result in enhanced expression of alpha-fetoprotein mRNA. However, the patterns of loss of DNA methylation at all sites examined after differentiation or 5-azacytidine treatment were remarkably similar, suggesting that the two occur by a similar mechanism, the inhibition of DNA methyltransferase activity. These results argue that the specificity for methylation loss at a given site is an inherent property of aggregated F9 cell chromatin. This system provides a model for studying a tissue-specific change in DNA methylation upon differentiation.

Citing Articles

Analysis of Hepatic Lentiviral Vector Transduction: Implications for Preclinical Studies and Clinical Gene Therapy Protocols.

Hu P, Hao Y, Tang W, Diering G, Zou F, Kafri T Viruses. 2025; 17(2).

PMID: 40007031 PMC: 11861806. DOI: 10.3390/v17020276.

Analysis of hepatic lentiviral vector transduction; implications for preclinical studies and clinical gene therapy protocols.

Hu P, Hao Y, Tang W, Diering G, Zou F, Kafri T bioRxiv. 2024; .

PMID: 39229157 PMC: 11370356. DOI: 10.1101/2024.08.20.608805.

Mechanisms Regulating Stemness and Differentiation in Embryonal Carcinoma Cells.

Kelly G, Gatie M Stem Cells Int. 2017; 2017:3684178.

PMID: 28373885 PMC: 5360977. DOI: 10.1155/2017/3684178.

Distinct DNA methylation patterns characterize differentiated human embryonic stem cells and developing human fetal liver.

Brunner A, Johnson D, Kim S, Valouev A, Reddy T, Neff N Genome Res. 2009; 19(6):1044-56.

PMID: 19273619 PMC: 2694474. DOI: 10.1101/gr.088773.108.

5-Methylcytosine DNA glycosylase participates in the genome-wide loss of DNA methylation occurring during mouse myoblast differentiation.

Jost J, Oakeley E, Zhu B, Benjamin D, Thiry S, Siegmann M Nucleic Acids Res. 2001; 29(21):4452-61.

PMID: 11691933 PMC: 60186. DOI: 10.1093/nar/29.21.4452.

References

Bouchard J, Momparler R . Incorporation of 5-Aza-2'-deoxycytidine-5'-triphosphate into DNA. Interactions with mammalian DNA polymerase alpha and DNA methylase. Mol Pharmacol. 1983; 24(1):109-14. View

Cremisi C . Effect of 5-azacytidine treatment on mouse embryonal carcinoma cells. J Cell Physiol. 1983; 116(2):181-90. DOI: 10.1002/jcp.1041160209. View

Mather E, PERRY R . Methylation status and DNase I sensitivity of immunoglobulin genes: changes associated with rearrangement. Proc Natl Acad Sci U S A. 1983; 80(15):4689-93. PMC: 384109. DOI: 10.1073/pnas.80.15.4689. View

CHARACHE S, Dover G, Smith K, Talbot Jr C, Moyer M, Boyer S . Treatment of sickle cell anemia with 5-azacytidine results in increased fetal hemoglobin production and is associated with nonrandom hypomethylation of DNA around the gamma-delta-beta-globin gene complex. Proc Natl Acad Sci U S A. 1983; 80(15):4842-6. PMC: 384141. DOI: 10.1073/pnas.80.15.4842. View

DOERFLER W . DNA methylation and gene activity. Annu Rev Biochem. 1983; 52:93-124. DOI: 10.1146/annurev.bi.52.070183.000521. View

Kratzer P, Chapman V, Lambert H, Evans R, Liskay R . Differences in the DNA of the inactive X chromosomes of fetal and extraembryonic tissues of mice. Cell. 1983; 33(1):37-42. DOI: 10.1016/0092-8674(83)90332-x. View

Santi D, Garrett C, Barr P . On the mechanism of inhibition of DNA-cytosine methyltransferases by cytosine analogs. Cell. 1983; 33(1):9-10. DOI: 10.1016/0092-8674(83)90327-6. View

Aviv H, Leder P . Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972; 69(6):1408-12. PMC: 426713. DOI: 10.1073/pnas.69.6.1408. View

Holliday R, Pugh J . DNA modification mechanisms and gene activity during development. Science. 1975; 187(4173):226-32. View

10.

Swartzendruber D, Lehman J . Neoplastic differentiation: interaction of simian virus 40 and polyoma virus with murine teratocarcinoma cells in vitro. J Cell Physiol. 1975; 85(2 Pt 1):179-87. DOI: 10.1002/jcp.1040850204. View

11.

Maniatis T, Jeffrey A, Kleid D . Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975; 72(3):1184-8. PMC: 432491. DOI: 10.1073/pnas.72.3.1184. View

12.

Martin G . Teratocarcinomas as a model system for the study of embryogenesis and neoplasia. Cell. 1975; 5(3):229-43. DOI: 10.1016/0092-8674(75)90098-7. View

13.

Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View

14.

Sherman M, Miller R . F9 embryonal carcinoma cells can differentiate into endoderm-like cells. Dev Biol. 1978; 63(1):27-34. DOI: 10.1016/0012-1606(78)90110-0. View

15.

Dziadek M, Adamson E . Localization and synthesis of alphafoetoprotein in post-implantation mouse embryos. J Embryol Exp Morphol. 1978; 43:289-313. View

16.

BIRNIE G . Isolation of nuclei from animal cells in culture. Methods Cell Biol. 1978; 17:13-26. DOI: 10.1016/s0091-679x(08)61131-0. View

17.

Waalwijk C, Flavell R . MspI, an isoschizomer of hpaII which cleaves both unmethylated and methylated hpaII sites. Nucleic Acids Res. 1978; 5(9):3231-6. PMC: 342244. DOI: 10.1093/nar/5.9.3231. View

18.

Strickland S, Mahdavi V . The induction of differentiation in teratocarcinoma stem cells by retinoic acid. Cell. 1978; 15(2):393-403. DOI: 10.1016/0092-8674(78)90008-9. View

19.

Constantinides P, Taylor S, Jones P . Phenotypic conversion of cultured mouse embryo cells by aza pyrimidine nucleosides. Dev Biol. 1978; 66(1):57-71. DOI: 10.1016/0012-1606(78)90273-7. View

20.

Tilghman S, Kioussis D, Gorin M, Ruiz J, Ingram R . The presence of intervening sequences in the alpha-fetoprotein gene of the mouse. J Biol Chem. 1979; 254(15):7393-9. View