title: Demethylation of RUNX3 by vincristine in colorectal adenocarcinoma cells.
abstract: BACKGROUND
  Methylation-mediated inactivation of tumor-suppressor genes is a critical event during the pathogenesis of many malignancies. Vincristine is a conventional anticancer drug used to treat various types of cancers. However, few studies describe the epigenetic-based effects of vincristine. In this study, changes in the methylation of runt-related transcription factor-3 (RUNX3) were investigated in CCD18Co normal colon cells and DLD-1 colorectal adenocarcinoma cells.
  
  
  METHODS
  CCD18Co and DLD-1 cells were treated with vincristine, and the methylation status was assessed using quantitative methylation-specific polymerase chain reaction (QMSP). Eleven normal colon tissues and 105 colorectal cancer tissues were investigated by methylation and mRNA expression of RUNX3 using QMSP and real-time reverse transcription polymerase chain reaction (real time-PCR).
  
  
  RESULTS
  RUNX3 was demethylated after vincristine treatment in DLD-1 cells. The expression of RUNX3 mRNA was down-regulated in DLD-1 cells because of DNA hypermethylation, but was restored after vincristine treatment. In addition, hypermethylation of RUNX3 was detected in 70 out of 105 colorectal carcinomas (66.7%). RUNX3 hypermethylation was greater in colon cancer tissues than in rectal cancer tissues. The expression of RUNX3 mRNA was reduced in 68 out of 105 colorectal cancer tissues (64.8%).
  
  
  CONCLUSIONS
  These results demonstrate that vincristine demethylates RUNX3 in colorectal adenocarcinoma cells, and restores its expression.
authors.affiliation: Institute of Human Genetics, Department of Anatomy, Korea University College of Medicine, 126-1, Anam-dong 5-ga, Seongbuk-gu, Seoul 136-705, Korea. parksh@korea.ac.kr.
authors.forename: Ji Wook
authors.initials: JW
authors.lastname: Moon
authors.forename: Soo Kyung
authors.initials: SK
authors.lastname: Lee
authors.forename: Jung Ok
authors.initials: JO
authors.lastname: Lee
authors.forename: Ji Hae
authors.initials: JH
authors.lastname: Kim
authors.forename: Nami
authors.initials: N
authors.lastname: Kim
authors.forename: Jin
authors.initials: J
authors.lastname: Kim
authors.forename: Hyeon Soo
authors.initials: HS
authors.lastname: Kim
authors.forename: Sun-Hwa
authors.initials: SH
authors.lastname: Park
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects*; D003106:Colon / Q000187:drug effects / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D050677:Core Binding Factor Alpha 3 Subunit / Q000235:genetics* / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008875:Middle Aged; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D014750:Vincristine / Q000494:pharmacology*; D055815:Young Adult
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D050677:Core Binding Factor Alpha 3 Subunit; D012333:RNA, Messenger; C493729:Runx3 protein, human; D014750:Vincristine
keywords: DNA methylation; RUNX3; colonic neoplasms; demethylation; methylation-specific polymerase chain reaction; vincristine