PMID- 18404682
OWN - NLM
STAT- MEDLINE
DCOM- 20080703
LR  - 20220311
IS  - 1097-0215 (Electronic)
IS  - 0020-7136 (Linking)
VI  - 123
IP  - 3
DP  - 2008 Aug 1
TI  - DNA methylation and histone modifications cause silencing of Wnt antagonist gene 
      in human renal cell carcinoma cell lines.
PG  - 535-42
LID - 10.1002/ijc.23514 [doi]
AB  - Secreted frizzled-related protein 2 (sFRP2) is a negative modulator of the 
      Wingless-type (Wnt) signaling pathway, and shown to be inactivated in renal cell 
      carcinoma (RCC). However, the molecular mechanism of silencing of sFRP2 is not 
      fully understood. Our study was designed to elucidate the silencing mechanism of 
      sFRP2 in RCC. Expression of sFRP2 was examined in 20 pairs of primary cancers by 
      immunohistochemistry. Kidney cell lines (HK-2, Caki-1, Caki-2, A-498 and ACHN) 
      were analyzed for sFRP2 expression using real-time RT-PCR and Western blotting. 
      The methylation status at 46 CpG sites of the 2 CpG islands in the sFRP2 promoter 
      was characterized by bisulfite DNA sequencing. Histone modifications were 
      assessed by chromatin immunoprecipitation (ChIP) assay using antibodies against 
      AcH3, AcH4, H3K4 and H3K9. sFRP2 was frequently repressed in primary cancers and 
      in RCC cells. The majority of sFRP2 negative cells had a methylated promoter. 
      Meanwhile, sFRP2 expression was repressed by a hypomethylated promoter in Caki-1 
      cells, and these cells had a repressive histone modification at the promoter. In 
      Caki-1 cells, sFRP2 was reactivated by trichostatin A (TSA). Repressive histone 
      modifications were also observed in RCC cells with hypermethylated promoters, but 
      sFRP2 was reactivated only by 5-aza-2'-deoxycytidine (DAC) and not by TSA. 
      However, the activation of the silenced sFRP2 gene could be achieved in all cells 
      using a combination of DAC and TSA. This is the first report indicating that 
      aberrant DNA methylation and histone modifications work together to silence the 
      sFRP2 gene in RCC cells.
FAU - Kawamoto, Ken
AU  - Kawamoto K
AD  - Department of Urology, Veterans Affairs Medical Center and University of 
      California School of Medicine, San Francisco, CA 94121, USA.
FAU - Hirata, Hiroshi
AU  - Hirata H
FAU - Kikuno, Nobuyuki
AU  - Kikuno N
FAU - Tanaka, Yuichiro
AU  - Tanaka Y
FAU - Nakagawa, Masayuki
AU  - Nakagawa M
FAU - Dahiya, Rajvir
AU  - Dahiya R
LA  - eng
GR  - R01CA101844/CA/NCI NIH HHS/United States
GR  - R01CA130860/CA/NCI NIH HHS/United States
GR  - R01CA111470/CA/NCI NIH HHS/United States
GR  - R01 CA130860/CA/NCI NIH HHS/United States
GR  - T32DK007790/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Int J Cancer
JT  - International journal of cancer
JID - 0042124
RN  - 0 (Antimetabolites, Antineoplastic)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Histone Deacetylase Inhibitors)
RN  - 0 (Histones)
RN  - 0 (Hydroxamic Acids)
RN  - 0 (Membrane Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (SFRP2 protein, human)
RN  - 0 (Wnt Proteins)
RN  - 3X2S926L3Z (trichostatin A)
RN  - 776B62CQ27 (Decitabine)
RN  - EC 2.1.1.- (DNA Modification Methylases)
RN  - M801H13NRU (Azacitidine)
SB  - IM
MH  - Antimetabolites, Antineoplastic/pharmacology
MH  - Apoptosis/genetics
MH  - Azacitidine/analogs & derivatives/pharmacology
MH  - Blotting, Western
MH  - Carcinoma, Renal Cell/genetics/*metabolism
MH  - Cell Line, Tumor
MH  - CpG Islands/genetics
MH  - *DNA Methylation
MH  - DNA Modification Methylases/antagonists & inhibitors
MH  - Decitabine
MH  - Enzyme Inhibitors/pharmacology
MH  - Gene Expression Regulation, Neoplastic/drug effects
MH  - *Gene Silencing/drug effects
MH  - Histone Deacetylase Inhibitors
MH  - Histones/*metabolism
MH  - Humans
MH  - Hydroxamic Acids/pharmacology
MH  - Immunohistochemistry/methods
MH  - Immunoprecipitation
MH  - Kidney Neoplasms/genetics/*metabolism
MH  - Membrane Proteins/*genetics
MH  - Promoter Regions, Genetic
MH  - RNA, Messenger/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Signal Transduction/drug effects
MH  - Wnt Proteins/*metabolism
EDAT- 2008/04/12 09:00
MHDA- 2008/07/04 09:00
CRDT- 2008/04/12 09:00
PHST- 2008/04/12 09:00 [pubmed]
PHST- 2008/07/04 09:00 [medline]
PHST- 2008/04/12 09:00 [entrez]
AID - 10.1002/ijc.23514 [doi]
PST - ppublish
SO  - Int J Cancer. 2008 Aug 1;123(3):535-42. doi: 10.1002/ijc.23514.