PMID- 17122150
OWN - NLM
STAT- MEDLINE
DCOM- 20070201
LR  - 20161124
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 47
IP  - 12
DP  - 2006 Dec
TI  - Activation of caspase-8 and caspase-12 pathways by 7-ketocholesterol in human 
      retinal pigment epithelial cells.
PG  - 5569-75
AB  - PURPOSE: To determine whether caspase or cathepsin pathways are activated in 
      human retinal pigment epithelial cells (ARPE-19) after exposure to 
      7-ketocholesterol (7kCh). METHODS: ARPE-19 cells were exposed to 7kCh with or 
      without z-VAD-fmk, a pan-caspase inhibitor. Caspase-3, -8, and -9 activities were 
      measured by a fluorochrome inhibitor of caspase (FLICA) assay. Caspase-12 
      activity was detected by Western blotting. RT-PCR was performed for 18s, 
      mortalin-2, cathepsins B, D, and L/V2. RESULTS: At 24 hours, 7kCh-treated 
      cultures had increased caspase-8 (P < 0.001) and caspase-3 (P < 0.001) activities 
      compared with vehicle-treated cultures. 7kCh-induced caspase-3 activation was 
      blocked by z-VAD-fmk (P < 0.001). Caspase-9 was not activated by 7kCh treatment 
      (P > 0.05). Procaspase-12 was cleaved into its active form after treatment with 
      7kCh for 24 hours. At 6 hours, the RNA level for mortalin-2, a pro-survival gene, 
      was upregulated. ARPE-19 cells did not express RNA for cathepsins B, D, or L/V2 
      under any conditions. CONCLUSIONS: In ARPE-19 cells, 7kCh-induced apoptosis uses 
      the receptor-mediated caspase-8 pathway and the endoplasmic reticulum 
      stress-induced caspase-12 pathway but not the mitochondrial caspase-9 pathway. 
      The cathepsin pathways are not involved in 7kCh-induced cell death. These data 
      demonstrate that 7kCh causes a loss of cell viability through caspase-dependent 
      apoptosis and can act as an oxidative stressor leading to retinal pigment 
      epithelial cell atrophy. Elucidating the specific apoptotic pathways involved may 
      have therapeutic potential for AMD and other retinal diseases.
FAU - Luthra, Saurabh
AU  - Luthra S
AD  - Department of Ophthalmology, School of Medicine, University of California, 
      Irvine, California 92868, USA.
FAU - Fardin, Babak
AU  - Fardin B
FAU - Dong, Joyce
AU  - Dong J
FAU - Hertzog, Dieter
AU  - Hertzog D
FAU - Kamjoo, Sami
AU  - Kamjoo S
FAU - Gebremariam, Simon
AU  - Gebremariam S
FAU - Butani, Virit
AU  - Butani V
FAU - Narayanan, Raja
AU  - Narayanan R
FAU - Mungcal, Jeanie K
AU  - Mungcal JK
FAU - Kuppermann, Baruch D
AU  - Kuppermann BD
FAU - Kenney, M Cristina
AU  - Kenney MC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Amino Acid Chloromethyl Ketones)
RN  - 0 (Cysteine Proteinase Inhibitors)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (HSP70 Heat-Shock Proteins)
RN  - 0 (HSPA9 protein, human)
RN  - 0 (Ketocholesterols)
RN  - 0 (Mitochondrial Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone)
RN  - EC 1.14.14.23 (Cholesterol 7-alpha-Hydroxylase)
RN  - EC 3.4.- (Cathepsins)
RN  - EC 3.4.22.- (CASP8 protein, human)
RN  - EC 3.4.22.- (Caspase 12)
RN  - EC 3.4.22.- (Caspase 3)
RN  - EC 3.4.22.- (Caspase 8)
RN  - O7676FE78M (7-ketocholesterol)
SB  - IM
MH  - Amino Acid Chloromethyl Ketones/pharmacology
MH  - Blotting, Western
MH  - Caspase 12/*metabolism
MH  - Caspase 3/metabolism
MH  - Caspase 8/*metabolism
MH  - Cathepsins/genetics/metabolism
MH  - Cell Line
MH  - Cholesterol 7-alpha-Hydroxylase/antagonists & inhibitors
MH  - Cysteine Proteinase Inhibitors/pharmacology
MH  - Enzyme Activation
MH  - Enzyme Inhibitors/*pharmacology
MH  - HSP70 Heat-Shock Proteins/genetics/metabolism
MH  - Humans
MH  - Ketocholesterols/*pharmacology
MH  - Mitochondrial Proteins
MH  - Pigment Epithelium of Eye/*drug effects/metabolism
MH  - RNA, Messenger/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Up-Regulation
EDAT- 2006/11/24 09:00
MHDA- 2007/02/03 09:00
CRDT- 2006/11/24 09:00
PHST- 2006/11/24 09:00 [pubmed]
PHST- 2007/02/03 09:00 [medline]
PHST- 2006/11/24 09:00 [entrez]
AID - 47/12/5569 [pii]
AID - 10.1167/iovs.06-0333 [doi]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2006 Dec;47(12):5569-75. doi: 10.1167/iovs.06-0333.