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Abstract
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Calumenin is a chaperon protein of endoplasmic reticulum and inhibitor of gamma-carboxylation system for vitamin K dependent (VKD) proteins that its knockdown is important for production of VKD proteins. The aim of this study was downregulation of calumenin by miRNA(s). In this regard, our in silico studies predicted has-miR-30d to target calumenin. Then, we selected this miRNA to knockdown this gene. Also, we take the advantage of a control negative miRNA which can’t target any gene of human to evaluate the effect of transfection of miRNA(s) on cells. Altogether, we added two different doses of miRNA(s) in to Hek293T cells. Subsequently, total RNA was extracted from cells at 48 hours post-transfection and subjected to cDNA synthesis. Down/up-regulation of calumenin transcripts was studied by Real-Time PCR and ∆∆Ct analysis. The Glyceraldehyde 3-phosphate dehydrogenase was used as housekeeping gene in this study. Our results showed that transfection of hsa-miR-30d resulted in 23% downregulation of calumenin transcripts, while transfection of half of the previous dose of hsa-miR-30d showed 16% downregulation. By the way, the expression level of calumenin reduced to 77% and 84% of the control cells in these tests. Whereas, the transfection of control negative miRNA showed no effect on calumenin knockdown. In conclusion, the results of this study demonstrated that hsa-miR-30d could specifically target and knockdown the calumenin transcripts and one could take the advantage of this miRNA to target human calumenin.
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