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MC Calonge, RM Corrales, V Saez, Y Diebold, A Enriquez de Salamanca, A Mayo, JM Herreras, FJ Chaves; Cyclosporin A Does Not Alter The Expression Of Ocular Mucin Genes In A Normal Human Conjunctival (NHC) Cell Line . Invest. Ophthalmol. Vis. Sci. 2002;43(13):50.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Human conjunctival epithelium in vivo expresses MUC1, 2, 4, 5AC and 7 mucin genes. We proved that our normal human conjunctival (NHC) cell line, of pure epithelial origin, expresses MUC1, 2, and 4 and does not express MUC5AC and 7. In addition, there is evidence that cyclosporin A (CsA) may modify the altered expression of mucin genes in dry eye conditions. However, there is no data regarding the potential effect of CsA on the expression of ocular mucin genes in healthy conditions. This study was designed to investigate the effect of a 0.05% CsA emulsion in the expression of these mucin genes on the NHC cell line. Methods: NHC cell line was cultured under three different conditions: 1) supplemented DMEM/F-12 culture medium, 2) supplemented DMEM/F-12 culture medium and 0.05% CsA emulsion, 3) supplemented DMEM/F-12 culture medium and vehicle. Just prior (basal conditions) and after 12, 24, 36, 48 and 60 hours of incubation, real time RT-PCR was performed using total RNA from all samples to examine the expression of MUC1, MUC2, MUC4, MUC5AC and MUC7 human mucin genes. The relative quantity of mucin genes was determined by SYBR® Green I dye detection. Amplification of one housekeeping gene (GAPDH) was performed to normalize the amount of expression levels. Results: MUC1, MUC2 and MUC4 were all detected in basal conditions (expression levels of MUC1 were 6.85 more times than MUC2 and 1.34 than MUC4) and at all time-points, with no significant differences. When CsA emulsion or vehicle was added to the culture media, MUC1, MUC2 and MUC4 levels remained unchanged. Amplification of MUC5AC and MUC7 was not detected at any time-point in any of the different culture conditions. Conclusion: The basal expression of MUC1, MUC2 and MUC4 mucin genes in our NHC cell line is not altered by a 0.05% CsA emulsion. More research is needed to determine how CsA can modify the expression of these genes in pathological conditions.
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