Fullerene–polyvinylpyrrolidone clathrate localizes in the cytoplasm to prevent Ultraviolet‐A ray‐induced DNA‐fragmentation and activation of the transcriptional factor NF‐kappaB

Abstract

By Western blot and immunostaining we proved that polyvinylpyrrolidone (PVP)-wrapped fullerene molecules (PVP-fullerene) could combine the 8- and 53-kb proteins which localize in the membrane of human skin keratinocytes HaCaT. Only fullerene molecules are able to cross the lipid membrane and conjugate 53-kb proteins in the cytosol. There are no fullerene molecules detectable in the nucleus or cytoskeleton. Ultraviolet-A (UVA)-irradiation on HaCaT or normal human epidermal melanocytes (NHEM) caused nuclear fragmentations, lowering of intracellular DNA-contents below diploidy, concurrently with the repressed DNA synthesis and the increased DNA-3'OH cleavage terminals, all of which were repressed by PVP-fullerene, as shown by flow cytometry and PI- or TUNEL-stain fluorography. Translocation of the transcriptional factor NF-kappaB in the cytoplasm to the nucleus of the keratinocytes was caused with UVA and repressed by PVP-fullerene with cytoprotective effects. Thus, the PVP-fullerene may be developed as a UV-protective agent with DNA-preservative effects owing to its combinative ability to molecules in the cytoplasm and cytomembrane, and then represses cellular oxidative stress and blocks abnormal signal pathways.