[Effects of 5-Aza-2'-deoxycitydine and trichostatin A on expression and methylation of CHFR in human laryngreal carcinoma cell line].
OBJECTIVE
To explore the effects of 5-Aza-2'-deoxycitydine(5-Aza-dC) and trichostatin A (TSA) on the expression and methylation of CHFR in human laryngeal carcinoma cell line.


METHODS
The mRNA expression and promoter hypermethylation and were detected by Realtime fluro-genetic quantitative PCR and methylation specific PCR in Hep-2 cell line, which were cultured in vitro and then treated with different concentrations of 5-Aza-dC and TSA.


RESULTS
Compared with the control team, 5-Aza-dC alone reactivated expression of the CHFR in Hep-2 cell line (1.75 +/- 0.21). TSA had no effect on gene expression (1.05 +/- 0.13). The combined treatment with 5-Aza-dC and TSA increased gene expression (2.15 +/- 0.18). The cell lines showed a characteristic DNA methylation status. 5-Aza-dC and combined 5-Aza-dC and TSA resulted in demethylation of CHFR. In contrast, TSA alone did not affect the DNA methylation status of CHFR.


CONCLUSIONS
Hypermethylation of CHFR gene promoter is a common event in the occurrence and development of laryngeal carcinoma. The promoter aberrant methylation of CHFR is a main cause for down-expression of CHFR. After either treatment with 5-Aza-dC alone or in combination with TSA, the expression of CHFR is up-regulated duo to the reversal methylation. It can be a new idea to the therapy of laryngeal carcinoma.
Department of Otorhinolaryngology, Shengjing Hospital of China Medical University, Shenyang 110004, China.
Lixia
L
He
Wenyue
W
Ji
Jing
J
Yang
Xudong
X
Zhao
D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation / Q000187:drug effects; D000077209:Decitabine; D015870:Gene Expression / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D007822:Laryngeal Neoplasms / Q000378:metabolism*; D008745:Methylation / Q000187:drug effects; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D000075223:Poly-ADP-Ribose Binding Proteins; D011401:Promoter Regions, Genetic; D044767:Ubiquitin-Protein Ligases
D018797:Cell Cycle Proteins; D006877:Hydroxamic Acids; D009363:Neoplasm Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; C012589:trichostatin A; D000077209:Decitabine; C412002:CHFR protein, human; D044767:Ubiquitin-Protein Ligases; D001374:Azacitidine