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Title

EFFECT OF HEAVY METALS ON SILENCING OF ENGINEERED LONG INTERSPERSED ELEMENT-1 RETROTRANSPOSON IN NONDIVIDING NEUROBLASTOMA CELL LINE

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 Start Page 171 | End Page 178

Abstract

 Background: L1 RETROTRANSPOSONs are the most active mobile DNA elements in human genome. Unregulated L1 retrotransposition may have deleterious effect by disrupting vital genes and inducing genomic instabilities.Therefore, human cells control L1 elements by silencing their activities through epigenetic mechanisms. It has been shown that CELL DIVISION and HEAVY METALS stimulate the frequency of L1 activities. Removal of silencing by L1 motivators may restart L1 element functions. Here, we have proposed that weather neurotoxic environmental HEAVY METALS (as L1 stimulating factors) have a role in removing L1 silencing and restating its activities in nondividing neuronal cells.Methods: L1-RP green fluorescent protein (GFP) -tagged knock-in human neuroblastoma clones were prepared. Single-cell clone was treated with mitomycin-c combined with nontoxic and toxic concentrations of iron (Fe), copper (Cu), and mercury (Hg). Silencing status of engineered L1 elements in dividing and nondividing cells was determined through measuring the amount of GFP expressing cells with flow cytometry. The cytotoxic effect of mitomycin-c combined with metals was measured by MTT assay.Results: Hg in nondividing cells and Fe, Cu, and Hg in dividing neuroblastoma cells could significantly remove L1 silencing. Also, mitomycin-c treatment did not have any effect on metal toxicity status in neuroblastoma cells.Conclusion: Totally, our findings have shown that CELL DIVISION has a role in removing L1 silencing as well as L1 retrotransposition induced by environmental HEAVY METALS. It has been also indicated that Hg at all concentrations could remove silencing of engineered L1 element regardless of cell cycle state. Iran.

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