title: Differential TERT promoter methylation and response to 5-aza-2'-deoxycytidine in acute myeloid leukemia cell lines: TERT expression, telomerase activity, telomere length, and cell death.
abstract: The catalytic subunit of human telomerase (TERT) is highly expressed in cancer cells, and correlates with complex cytogenetics and disease severity in acute myeloid leukemia (AML). The TERT promoter is situated within a large CpG island, suggesting that expression is methylation-sensitive. Studies suggest a correlation between hypermethylation and TERT overexpression. We investigated the relationship between TERT promoter methylation and expression and telomerase activity in human leukemia and lymphoma cell lines. DAC-induced demethylation and cell death were observed in all three cell lines, as well as telomere shortening in HL-60 cells. DAC treatment reduced TERT expression and telomerase activity in OCI/AML3 and HL-60 cells, but not in U937 cells. Control U937 cells expressed lower levels of TERT mRNA, carried a highly methylated TERT core promoter, and proved more resistant to DAC-induced repression of TERT expression and cell death. AML patients had significantly lower methylation levels at several CpGs than "well elderly" individuals. This study, the first to investigate the relationship between TERT methylation and telomerase activity in leukemia cells, demonstrated a differential methylation pattern and response to DAC in three AML cell lines. We suggest that, although DAC treatment reduces TERT expression and telomerase activity, this is unlikely to occur via direct demethylation of the TERT promoter. However, further investigations on the regions spanning CpGs 7-12 and 14-16 may reveal valuable information regarding transcriptional regulation of TERT.
authors.affiliation: Centre for Cancer Research and Cell Biology, Queens University Belfast, Belfast BT9 7BL, UK. kap6@st-andrews.ac.uk
authors.forename: Kerry A
authors.initials: KA
authors.lastname: Pettigrew
authors.forename: Richard N
authors.initials: RN
authors.lastname: Armstrong
authors.forename: Hilary A A
authors.initials: HA
authors.lastname: Colyer
authors.forename: Shu-Dong
authors.initials: SD
authors.lastname: Zhang
authors.forename: Irene Maeve
authors.initials: IM
authors.lastname: Rea
authors.forename: Rhiannon E
authors.initials: RE
authors.lastname: Jones
authors.forename: Duncan M
authors.initials: DM
authors.lastname: Baird
authors.forename: Ken I
authors.initials: KI
authors.lastname: Mills
mesh_terms: D000369:Aged, 80 and over; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D018899:CpG Islands; D019175:DNA Methylation; D000077209:Decitabine; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D018922:HL-60 Cells; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008297:Male; D011401:Promoter Regions, Genetic; D019098:Telomerase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D016615:Telomere / Q000235:genetics*; D020298:U937 Cells
chemical_list: D000964:Antimetabolites, Antineoplastic; D000077209:Decitabine; C509186:TERT protein, human; D019098:Telomerase; D001374:Azacitidine