Estrogen signaling in livers of male mice with hepatocellular carcinoma induced by exposure to arsenic in utero.
BACKGROUND
Exposure of pregnant mice to inorganic arsenic induces a spectrum of tumors, including hepatocellular carcinoma (HCC), in their adult offspring similar to that induced by exposing adult mice to estrogenic compounds. To investigate whether arsenic exposure in utero causes altered estrogen signaling, we examined expression of estrogen receptor-alpha (ER-alpha), cyclin D1 (an estrogen-responsive hepatic oncogene), and several cytochrome P450 genes (with sexually dimorphic liver expression patterns) in livers from adult male mice with in utero arsenic-induced HCC.


METHODS
Quantitative real-time reverse transcription-polymerase chain reaction was used to evaluate gene expression in livers of adult male mice that had (i.e., exposed mice; n = 8) or had not (i.e., control mice; n = 5) been exposed to arsenic in utero. DNA methylation status of portions of the ER-alpha and cyclin D1 gene promoters in liver tissue was measured using methylation-specific polymerase chain reaction. Statistical tests were two-sided.


RESULTS
ER-alpha mRNA levels were 3.1-fold (95% confidence interval [CI] = 2.0-fold to 4.3-fold) higher in livers of exposed mice than in those of control mice, and cyclin D1 levels were 3.0-fold (95% CI = 1.7-fold to 4.3-fold) higher. Exposed mice showed a feminized expression pattern of several cytochrome P450 genes, expressing the female-dominant CYP2A4 (P =.017 versus control) and CYP2B9 (P<.001) genes at 8.7 and 10.5 times, respectively, the level in control mice and expressing the male-dominant CYP7B1 at approximately one-fourth the level in control mice(P =.0012). Exposed mice exhibited reduced (by approximately 90%) methylation of the ER-alpha gene promoter in liver DNA as compared with control mice; the cyclin D1 gene promoter was not methylated in either exposed or control mice.


CONCLUSIONS
Altered estrogen signaling may play a role in induction of HCC by arsenic exposure in utero. Specifically, overexpression of ER-alpha, potentially through promoter region hypomethylation, in livers of such mice may be linked to the hepatocarcinogenicity of arsenic.
Waalkes
Michael P
MP
Inorganic Carcinogenesis Section, Laboratory of Comparative Carcinogenesis, National Cancer Institute at National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA. waalkes@niehs.nih.gov
Liu
Jie
J
Chen
Hua
H
Xie
Yaxiong
Y
Achanzar
William E
WE
Zhou
Yun-Su
YS
Cheng
Min-Liang
ML
Diwan
Bhalchandra A
BA
D000818:Animals; D001152:Arsenicals; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D002273:Carcinogens; D056486:Chemical and Drug Induced Liver Injury; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D000072467:Cytochrome P450 Family 2; D000072474:Cytochrome P450 Family 7; D019175:DNA Methylation; D004781:Environmental Exposure / Q000009:adverse effects; D047628:Estrogen Receptor alpha; D004967:Estrogens / Q000378:metabolism*; D005333:Fetus; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008107:Liver Diseases / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental / Q000139:chemically induced / Q000378:metabolism*; D008297:Male; D051379:Mice; D012333:RNA, Messenger / Q000378:metabolism; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013250:Steroid Hydroxylases / Q000378:metabolism
D001152:Arsenicals; D002273:Carcinogens; D047628:Estrogen Receptor alpha; D004967:Estrogens; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D019938:Cyclin D1; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; C115867:oxysterol 7-alpha-hydroxylase; D001189:Aryl Hydrocarbon Hydroxylases; C460223:Cyp2a4 protein, mouse; C121503:Cyp2b9 protein, mouse; D000072467:Cytochrome P450 Family 2; D000072474:Cytochrome P450 Family 7; C499571:CYP7B1 protein, human