title: Epigenetic inactivation of TFPI-2 as a common mechanism associated with growth and invasion of pancreatic ductal adenocarcinoma.
abstract: Using microarrays, we have screened for genes reactivated by drugs that modify epigenetic mechanisms in pancreatic cancer cells. One of the genes identified was tissue factor pathway inhibitor 2 (TFPI-2), which encodes for a broad-spectrum serine proteinase inhibitor that negatively regulates the extracellular matrix degradation, an essential step in tumor invasion and metastasis. We therefore investigated the expression and methylation patterns of the TFPI-2 gene in pancreatic adenocarcinoma, and determined its role in tumor growth and invasion. In contrast to its abundant expression in normal pancreas, TFPI-2 mRNA was undetectable in a high fraction of pancreatic cancer cell lines and in primary pancreatic ductal neoplasms (IPMNs). Loss of TFPI-2 expression was associated with aberrant hypermethylation of its promoter CpG island. Treatment with the phorbol ester (PMA), known to stimulate the TFPI-2 promoter activity, augmented the TFPI-2 expression in cell lines with unmethylated or partially methylated TFPI-2, but failed to induce the expression in cell lines that harbored fully methylated TFPI-2. Aberrant methylation of TFPI-2 was also detected in 73% (102/140) of pancreatic cancer xenografts and primary pancreatic adenocarcinomas, was more likely in older patients with pancreatic cancer, and significantly correlated with progression of IPMNs (P=0.0002). Restored expression of the TFPI-2 gene in nonexpressing pancreatic cancer cells resulted in marked suppression in their proliferation, migration, and invasive potential in vitro. We thus conclude that epigenetic inactivation of TFPI-2 is a common mechanism that contributes to the aggressive phenotype of pancreatic ductal adenocarcinoma.
authors.affiliation: Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21205, USA.
authors.forename: Norihiro
authors.initials: N
authors.lastname: Sato
authors.forename: Antony R
authors.initials: AR
authors.lastname: Parker
authors.forename: Noriyoshi
authors.initials: N
authors.lastname: Fukushima
authors.forename: Yohei
authors.initials: Y
authors.lastname: Miyagi
authors.forename: Christine A
authors.initials: CA
authors.lastname: Iacobuzio-Donahue
authors.forename: James R
authors.initials: JR
authors.lastname: Eshleman
authors.forename: Michael
authors.initials: M
authors.lastname: Goggins
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000601:surgery; D000964:Antimetabolites, Antineoplastic / Q000633:toxicity; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000633:toxicity*; D001483:Base Sequence; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics* / Q000601:surgery; D045744:Cell Line, Tumor; D017931:DNA Primers; D000077209:Decitabine; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D006801:Humans; D006877:Hydroxamic Acids / Q000633:toxicity; D019656:Loss of Heterozygosity; D020411:Oligonucleotide Array Sequence Analysis; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000601:surgery; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014183:Transplantation, Heterologous
chemical_list: D000964:Antimetabolites, Antineoplastic; D017931:DNA Primers; D006023:Glycoproteins; D006877:Hydroxamic Acids; C021240:tissue-factor-pathway inhibitor 2; C012589:trichostatin A; D000077209:Decitabine; D001374:Azacitidine