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Depletion of SLC4A11 causes cell death by apoptosis in an immortalized human corneal endothelial cell line.
- Source :
-
Investigative ophthalmology & visual science [Invest Ophthalmol Vis Sci] 2012 Jun 05; Vol. 53 (7), pp. 3270-9. Date of Electronic Publication: 2012 Jun 05. - Publication Year :
- 2012
-
Abstract
- Purpose: To investigate the effects of SLC4A11 gene depletion in human corneal endothelial cells.<br />Methods: To achieve stable downregulation of SLC4A11 gene expression in immortalized human corneal endothelial cells (HCECs), short-hairpin RNA (shRNA) targeted against SLC4A11 was used. Cell growth and viability were determined using the real-time cell analyzer and trypan blue staining respectively. Apoptosis was investigated by Annexin V and TUNEL assays. Alterations in apoptotic gene expression following SLC4A11 silencing were determined using the RT(2)Profiler PCR array for human apoptosis while activation of the apoptotic pathway was ascertained by western analysis.<br />Results: SLC4A11 silencing in HCECs could be achieved by stable expression of shRNA targeted against SLC4A11. SLC4A11 knockdown suppressed HCEC growth and reduced HCEC viability compared to the control. This reduction in cell growth is associated with increased apoptosis in SLC4A11-silenced cells.<br />Conclusions: Our data suggest that the reduction of cell number with time in SLC4A11-depleted HCECs is due to an increase in cell death by apoptosis. This suggests that SLC4A11 is necessary for cell survival and may explain the pathologic corneal endothelial cell loss in endotheliopathies due to SLC4A11 mutations.
- Subjects :
- Anion Transport Proteins biosynthesis
Antiporters biosynthesis
Apoptosis genetics
Blotting, Western
Cell Count
Cell Line
Cell Proliferation
Cell Survival
Endothelium, Corneal pathology
Gene Silencing
Humans
Immunohistochemistry
In Situ Nick-End Labeling
Mutation
RNA, Small Interfering biosynthesis
Reverse Transcriptase Polymerase Chain Reaction
Anion Transport Proteins genetics
Antiporters genetics
Cell Death genetics
Down-Regulation genetics
Endothelium, Corneal metabolism
Gene Expression Regulation
RNA, Messenger genetics
RNA, Small Interfering genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1552-5783
- Volume :
- 53
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Investigative ophthalmology & visual science
- Publication Type :
- Academic Journal
- Accession number :
- 22447871
- Full Text :
- https://doi.org/10.1167/iovs.11-8724