Glycine-N methyltransferase expression in HepG2 cells is involved in methyl group homeostasis by regulating transmethylation kinetics and DNA methylation

Yi Cheng Wang, Feng Yao Tang, Shih Yin Chen, Yi Ming Chen, En Pei Isabel Chiang

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Glycine-N methyltransferase (GNMT) is a potential tumor suppressor that is commonly inactivated in human hepatoma. We systematically investigated how GNMT regulates methyl group kinetics and global DNA methylation. HepG2 cells (GNMT inactive, GNMT-) and cells transfected with GNMT expressed vector (GNMT+) were cultured in low (10 μmol/L), adequate (100 μmol/L), or high (500 μmol/L) L-methionine, each with 2.27 μmol/L folate. Transmethylation kinetics were studied using stable isotopic tracers and GC-MS. Methylation status was determined by S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) levels, SAM:SAH ratio, DNA methyltransferase (DNMT) activity, and methylated cytidine levels in DNA. Compared with GNMT- cells, GNMT+ cells had lower homocysteine and greater cysteine concentrations. GNMT expression increased methionine clearance by inducing homocysteine transsulfuration and remethylation metabolic fluxes when cells were cultured in high or adequate L-methionine. In contrast, homocysteine remethylation flux was lower in GNMT+ cells than in GNMT- cells and homocysteine transsulfuration fluxes did not differ when cells were cultured in low methionine, suggesting that normal GNMT function helps to conserve methyl groups. Furthermore, GNMT expression decreased SAM and increased SAH levels and reduced DNMT activity in high or adequate, but not low, methionine cultures. In low methionine cultures, restoring GNMT in HepG2 cells did not lead to sarcosine synthesis, which would waste methyl groups. Methylated cytidine levels were significantly lower in GNMT- cells than in GNMT+ cells. In conclusion, we have shown that GNMT affects transmethylation kinetics and SAM synthesis and facilitates the conservation of methyl groups by limiting homocysteine remethylation fluxes.

Original languageEnglish
Pages (from-to)777-782
Number of pages6
JournalJournal of Nutrition
Volume141
Issue number5
DOIs
Publication statusPublished - May 1 2011
Externally publishedYes

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Glycine N-Methyltransferase
Hep G2 Cells
DNA Methylation
Homeostasis
Methionine
Homocysteine
S-Adenosylmethionine
S-Adenosylhomocysteine
Cytidine
Methyltransferases
Cultured Cells
DNA
Sarcosine

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Nutrition and Dietetics

Cite this

Glycine-N methyltransferase expression in HepG2 cells is involved in methyl group homeostasis by regulating transmethylation kinetics and DNA methylation. / Wang, Yi Cheng; Tang, Feng Yao; Chen, Shih Yin; Chen, Yi Ming; Chiang, En Pei Isabel.

In: Journal of Nutrition, Vol. 141, No. 5, 01.05.2011, p. 777-782.

Research output: Contribution to journalArticle

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