Depletion of SLC4A11 causes cell death by apoptosis in an immortalized human corneal endothelial cell line
| Autor: | Anandalakshmi Venkatraman, Eranga N. Vithana, Tin Aung, Li-Fong Seet, Jun Liu, Li Wei Koh, Rajiv R. Mohan, Divya Venkataraman, Joseph A. Bonanno, Jeppe Praetorius |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Programmed cell death
genetic structures Cell Survival Anion Transport Proteins Blotting Western Down-Regulation Apoptosis Cell Count Biology Antiporters Cell Line Downregulation and upregulation Cornea medicine In Situ Nick-End Labeling Gene silencing Humans Gene Silencing RNA Messenger RNA Small Interfering Cell Proliferation Regulation of gene expression Cell Death Cell growth Reverse Transcriptase Polymerase Chain Reaction Endothelium Corneal Articles Immunohistochemistry eye diseases Cell biology medicine.anatomical_structure Gene Expression Regulation Cell culture Mutation sense organs |
| Zdroj: | Liu, J, Seet, L-F, Koh, L W, Venkatraman, A, Venkataraman, D, Mohan, R R, Prætorius, J, Bonanno, J A, Aung, T & Vithana, E N 2012, ' Depletion of SLC4A11 causes cell death by apoptosis in an immortalized human corneal endothelial cell line ', Investigative Ophthalmology & Visual Science, vol. 53, no. 7, pp. 3270-9 . https://doi.org/10.1167/iovs.11-8724 |
| DOI: | 10.1167/iovs.11-8724 |
| Popis: | To investigate the effects of SLC4A11 gene depletion in human corneal endothelial cells.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.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.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. |
| Databáze: | OpenAIRE |
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