title: Methylated Bone Morphogenetic Protein 3 (BMP3) Gene: Evaluation of Tumor Suppressor Function and Biomarker Potential in Biliary Cancer.
abstract: BACKGROUND
  Although cholangiocarcinoma (CC) is an uncommon and highly lethal malignancy, early detection enables the application of potentially curative therapies and improves survival. Consequently, tools to improve the early diagnosis of CC are urgently needed. During a screen for genes epigenetically suppressed by methylation in CC that might serve as methylation markers for CC, we found that theBMP3gene is methylated in CC cell lines, but the potential diagnostic value and the function ofBMP3in CC are unknown.
  
  
  METHODS
  We aimed to quantitatively assessBMP3methylation in resected CC tumor specimens using methylation specific PCR and evaluate the tumor suppressor role ofBMP3in biliary cancer cell lines in comparison to an immortalized normal cholangiocyte cell line. Expression of BMP3 was quantified by mRNA levels before and after treatment with 5-Aza-2'-deoxycytidine and trichostatin A. After transfection with aBMP3-containing plasmid, cell viability was measured using the bromodeoxyuridine incorporation assay and apoptosis quantified by caspase assay.
  
  
  RESULTS
  In primary CC tumor tissue specimens significantly more methylatedBMP3copies were found when compared to matched benign bile duct epithelium from the same patient, with high specificity.BMP3expression was absent in cell lines withBMP3methylation; this suppression ofBMP3expression was reversed by treatment with a DNA demethylating agent and histone de-acetylase inhibitor. Transfection of aBMP3-expressing construct into aBMP3-negative biliary cancer cell line restoredBMP3mRNA expression and reduced cell proliferation and cell viability while increasing the rate of apoptosis.
  
  
  CONCLUSIONS
  These findings strongly support a tumor suppressor role forBMP3in CC and suggest thatBMP3methylation may be a new biomarker for early detection of CCs. of the peptidome are also involved.
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: John B
authors: JB
authors: Kisiel
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Jia
authors: J
authors: Li
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Hongzhi
authors: H
authors: Zou
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Abdul M
authors: AM
authors: Oseini
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Benjamin B
authors: BB
authors: Strauss
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Kadra H
authors: KH
authors: Gulaid
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Catherine D
authors: CD
authors: Moser
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Ileana
authors: I
authors: Aderca
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: David A
authors: DA
authors: Ahlquist
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Lewis R
authors: LR
authors: Roberts
authors: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester Minnesota, USA.
authors: Abdirashid M
authors: AM
authors: Shire
keywords: Biological markers; Cholangiocarcinoma; Early detection of cancer; Gallbladder neoplasms