Class I HDACs are mediators of smoke carcinogen-induced stabilization of DNMT1 and serve as promising targets for chemoprevention of lung cancer.
DNA methylation is an early event in bronchial carcinogenesis and increased DNA methyltransferase (DNMT)1 protein expression is a crucial step in the oncogenic transformation of epithelia. Here, we investigate the role of class I histone deacetylases (HDAC) 1 to 3 in the stabilization of DNMT1 protein and as a potential therapeutic target for lung cancer chemoprevention. Long-term exposure of immortalized bronchial epithelial cells (HBEC-3KT) to low doses of tobacco-related carcinogens led to oncogenic transformation, increased HDAC expression, cell-cycle independent increased DNMT1 stability, and DNA hypermethylation. Overexpression of HDACs was associated with increased DNMT1 stability and knockdown of HDACs reduced DNMT1 protein levels and induced DNMT1 acetylation. This suggests a causal relationship among increased class I HDACs levels, upregulation of DNMT1 protein, and subsequent promoter hypermethylation. Targeting of class I HDACs with valproic acid (VPA) was associated with reduced HDAC expression and a profound reduction of DNMT1 protein level. Treatment of transformed bronchial epithelial cells with VPA resulted in reduced colony formation, demethylation of the aberrantly methylated SFRP2 promoter, and derepression of SFRP2 transcription. These data suggest that inhibition of HDAC activity may reverse or prevent carcinogen-induced transformation. Finally, immunohistochemistry on human lung cancer specimens revealed a significant increase in DNMT1, HDAC1, HDAC2, and HDAC3 expression, supporting our hypotheses that class I HDACs are mediators of DNMT1 stability. In summary, our study provides evidence for an important role of class I HDACs in controlling the stability of DNMT1 and suggests that HDAC inhibition could be an attractive approach for lung cancer chemoprevention.
Atlanta VAMC, Winship Cancer Institute, Atlanta, GA 30322. johann.brandes@emory.edu.
Seth A
SA
Brodie
Ge
G
Li
Adam
A
El-Kommos
Hyunseok
H
Kang
Suresh S
SS
Ramalingam
Madhusmita
M
Behera
Khanjan
K
Gandhi
Jeanne
J
Kowalski
Gabriel L
GL
Sica
Fadlo R
FR
Khuri
Paula M
PM
Vertino
Johann C
JC
Brandes
D002274:Carcinogens, Environmental; D002283:Carcinoma, Bronchogenic / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control*; D002471:Cell Transformation, Neoplastic / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D018890:Chemoprevention / Q000379:methods; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism*; D056284:Histone Deacetylase 1 / Q000037:antagonists & inhibitors* / Q000502:physiology*; D056572:Histone Deacetylase Inhibitors / Q000627:therapeutic use*; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control*; D058990:Molecular Targeted Therapy; D011401:Promoter Regions, Genetic / Q000187:drug effects; D055550:Protein Stability / Q000187:drug effects; D012906:Smoke
D002274:Carcinogens, Environmental; D056572:Histone Deacetylase Inhibitors; D012906:Smoke; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human; C110550:HDAC1 protein, human; D056284:Histone Deacetylase 1