[Reversion transcriptional expression of DAPK in bladder cancer T24 cells 5-aza-2'-deoxycytidine].
OBJECTIVE
To investigate the methylation status of the promoter of resion death associated protein kinase (DAPK) gene in bladder cancer cell (T24), and study the effect of 5-aza-2'-deoxycytidine (5-aza-dc) on DAPK gene reactive expression in T24 and its inhibitory effect on T24.


METHODS
The bladder cancer cell T24 was treated with different doses of 5-aza-dc. The inhibitory effect and apoptosis rate were detected by MTT and flow cytometry, and the changes of DAPK mRNA and protein expression and the methylation status of DAPK promoter were assessed by RT-PCR, Western blotting, and methylation specific PCR, respectively.


RESULTS
The growth of bladder cancer cell was inhibited significantly and the maximal apoptosis rate detected by flow cytometry was (24.12-/+1.4)%. DAPK mRNA was not expressed in bladder cancer cell T24 in normal conditions. DAPK mRNA and protein re-expressed after 5-aza-dc (12.5 micromol/L) treatment in cell line T24 for 24 h, and DAPK promoter became unmethylated.


CONCLUSIONS
The promoter methylation can be an important factor for silencing the expression of DAPK in bladder cancer cell. 5-aza-dc can inhibit the growth and induce apoptosis of bladder cancer cells through reversing unmethylation status of DAPK promoter.
Department of Urology, Zhujiang Hospital, Southern Medical University, Guangzhou, China. xuningru@163.com
Ning-ru
NR
Xu
Chun-xiao
CX
Liu
Shao-bo
SB
Zheng
Hu-lin
HL
Li
Ya-wen
YW
Xu
Kai
K
Xu
D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors; D064550:Death-Associated Protein Kinases; D000077209:Decitabine; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects*; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000473:pathology
D051017:Apoptosis Regulatory Proteins; D012333:RNA, Messenger; D000077209:Decitabine; D015254:DNA Modification Methylases; D064550:Death-Associated Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D001374:Azacitidine