Pancreatic cancer DNMT1 expression and sensitivity to DNMT1 inhibitors.
DNA methyltransferase I (DNMT1) is the major methyltransferase responsible for methylating DNA and is overexpressed in many cancers. DNMT1 is also a therapeutic target for chemotherapy and chemoprevention. We hypothesized that loss of DNMT1 copy number could result in reduced DNMT1 levels and greater sensitivity to DNMT1 inhibitors. We examined DNMT1 expression in pancreatic cancers by immunohistochemistry and western blotting. We also examined DNMT1 copy number in 20 pancreatic cancer cell lines using Affymetrix SNP arrays and correlated copy number with DNMT1 expression. We tested 8 pancreatic cancer cell lines with DNMT1 inhibitors and measured growth inhibition. We identified overexpression of DNMT1 relative to normal pancreatic duct in 78.7% of pancreatic cancers (37/47) by immunohistochemistry and in 16/20 pancreatic cancer cell lines by western blot. Pancreatic cancer cell lines with loss of DNMT1 alleles tended to have lower DNMT1 expression (3 of 9 cell lines) compared to those without DNMT1 copy number loss (1 of 11). 5-aza-deoxycytidine (5-Aza-dC) treatment (1-10 uM) depleted DNMT1 in 7 of 8 pancreatic cancer cell lines. Three of four pancreatic cancers cell lines with low/normal DNMT1 expression were sensitive to growth inhibition by low dose 5-Aza-dC (1 uM), whereas only 1 of 4 cell lines with high DNMT1 expression had growth inhibition, and this occurred without evidence of DNMT1 depletion suggesting a different mechanism for growth inhibition in this cell line. Loss of DNMT1 alleles may reduce DNMT1 levels in some pancreatic cancers. Pancreatic cancers with low DNMT1 expression tend to be more sensitive to low-dose 5-azadeoxycytidine.
Li
Ang
A
Department of Pathology, The Johns Hopkins University School of Medicine.
Omura
Noriyuki
N
Department of Pathology, The Johns Hopkins University School of Medicine.
Hong
Seung-Mo
SM
Department of Pathology, The Johns Hopkins University School of Medicine.
Goggins
Michael
M
The Sol Goldman Center for Pancreatic Cancer Research.
D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D002392:Catechin / Q000031:analogs & derivatives / Q000494:pharmacology; D045744:Cell Line, Tumor / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002888:Chromosomes, Human, Pair 19 / Q000235:genetics; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D000077209:Decitabine; D019008:Drug Resistance, Neoplasm; D018628:Gene Dosage; D006801:Humans; D009154:Mutation; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins / Q000235:genetics
C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D000077209:Decitabine; D002392:Catechin; C045651:epigallocatechin gallate; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins; D001374:Azacitidine