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E. N. Vithana, A. Venkatraman, L. Koh, R. R. Mohan, L. Seet; Analysis of the Effects of SLC4A11 (CHED2) Gene Depletion on Corneal Endothelial Cell Growth and Proliferation. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1799.
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- SLC4A11 gene was successfully depleted at 10nM concentration by the two siRNAs at mRNA level. The HA-tagged SLC4A11 protein produced from a plasmid co-transfected with siRNA was also shown to be depleted by western analysis. The siRNA treatment for 48, 72 or 96 h appeared to have a stimulatory effect on cell proliferation and growth, although mRNA levels showed >95% depletion at all time points, compared to cells treated with negative siRNA control. SLC4A11 depletion also reduced the expression of pro-apoptotic factor Bax by about 50% and produced a negative result on TUNEL assay. Borate concentrations (1uM-1mM) were found not to significantly induce CEC growth compared to untreated cells. Concentrations 5-10mM were found to be slightly toxic to CECs.Conclusion- The effect of SLC4A11 depletion in transformed corneal endothelial cells differs from that observed in HeLa cells, where transient gene depletion reduced cell growth and proliferation. Work is in progress to test the effects of stable knockdown of SLC4A11. It is possible that NaBC1 has a different function in CECs and needs further investigation.
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