ShRNA-mediated gene silencing of AHR promotes the differentiation of             P19 mouse embryonic carcinoma cells into cardiomyocytes.
The aryl hydrocarbon receptor (AHR) is a basic helix-loop-helix (bHLH) transcription             factor that is activated by environmental contaminants including polychlorinated             biphenyls (PCBs). The AHR affects a variety of processes that are involved in             cell growth and differentiation. In this study, we constructed a P19 embryonic             carcinoma cell line with AHR gene silencing using the vector-based approach of             short hairpin (sh)RNA interference that allows cells to differentiate into cardiac             myocytes when treated with dimethyl sulfoxide (DMSO). The expression levels of             the cardiac development-specific GATA4 and Nkx2.5 genes were measured using real-time             quantitative polymerase chain reaction (qPCR). Our data showed that the expression             levels of the GATA4 and Nkx2.5 genes were increased in the AHR-silenced P19 cells             compared with the control groups. Four critical genes (ARNT, CYP1A1, GSK3β and             β-catenin) expressed in the AHR and in the Wnt signaling pathway were also measured             by qPCR. We found that the expression levels of ARNT, CYP1A1 and β-catenin were             suppressed, whereas GSK3β expression was elevated in the AHR-silenced P19 cells.             Therefore, it is possible that the silencing of AHR promotes the differentiation             of P19 cells through the AHR and Wnt signal transduction pathway.
State Key Laboratory of Reproductive Medicine, Department of Pediatrics, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing 210029, PR China.
Chun
C
Zhu
Yu-Lin
YL
Chen
Xue-Jie
XJ
Wang
Xiao-Shan
XS
Hu
Zhang-Bin
ZB
Yu
Shu-Ping
SP
Han
D000818:Animals; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D054278:Embryonal Carcinoma Stem Cells / Q000166:cytology* / Q000378:metabolism; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D018336:Receptors, Aryl Hydrocarbon / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
C497397:Arnt protein, mouse; D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C484469:Nkx2-5 protein, mouse; D034741:RNA, Small Interfering; D018336:Receptors, Aryl Hydrocarbon; D014157:Transcription Factors; D051176:beta Catenin; D051784:Aryl Hydrocarbon Receptor Nuclear Translocator; D019363:Cytochrome P-450 CYP1A1; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D038362:Glycogen Synthase Kinase 3; D004121:Dimethyl Sulfoxide