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Forum Kayastha, Darshini Ganatra, Alpesh Patel, Hardik Madhu, Abhay Vasavada; Mimosine suppresses proliferation and epithelial mesenchymal transition in lens epithelial cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3048.
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To evaluate the effect of mimosine on proliferation and epithelial mesenchymal transition (EMT) of lens epithelial cells.
Fetal human lens epithelial cell line (FHL124) was cultured in EMEM with 10% FBS. Cell viability and proliferation was evaluated by MTT assay. The proportion of cells at different phases of cell cycle was monitored by flow cytometer by propidium iodide staining. To evaluate the effect of growth factors (TGF-β2 and bFGF) induced EMT cells were divided into following groups, control-no treatment, treated with growth factors, treatment with mimosine alone, co-treatment of mimosine and growth factors for 24 hours (n=3). Epithelial marker, Pax6, EMT marker, alpha smooth muscle actin (α-SMA) and extracellular matrix marker, fibronectin were studied by immunofluorescence, immunoblotting and qPCR. Statistical analysis was carried out using student’s t-test and values with p<0.05 were considered significant.
IC 50 value of mimosine was 2mM. Cell proliferation was attenuated in dose and time dependent manner. Cell cycle analysis showed that mimosine arrests cells in G0/G1 phase in dose and time dependent manner. Protein and m-RNA level of pax6 significantly decreases with treatment of growth factors while it was significantly regained after co-treatment of mimosine. α-SMA and fibronectin were significantly down-regulated after co-treatment of mimosine when compared to growth factors treated group. Immunofluorescent staining of α-SMA and fibronectin was enhanced with treatment of growth factors while mimosine co-treatment weakened staining intensity.
Mimosine inhibits proliferation and growth factors induced EMT in lens epithelial cells by cell cycle arrest and modulation of EMT markers.
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