ACCUMULATION OF PLATINUM IN VESICLES FROM ATP7A-EXPRESSING CELLS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Recent studies have suggested a role for copper transporters in the transport of cisplatin. Altered expression of some copper transporters (ATP7A, ATP7B and CTR1) have led to altered accumulation of and sensitivity to cisplatin and other platinum-containing chemotherepeutic drugs. We have been investigating platinum accumulation and sensitivity using fibroblasts obtained from Menkes patients who lack ATP7A expression, Me32a, and the same cell line molecularly engineered to over-express ATP7A, MeMNK. Our current studies demonstrate higher whole cell and vesicular accumulation of platinum in cisplatin-treated MeMNK cells compared with non-expressing Me32a cells. While these results suggest a role for ATP7A in accumulation of platinum, we would like a more direct approach to determine whether specifically the vesicles that contain ATP7A are also the ones that accumulate the platinum in treated cells. Update: The vesicles that have ATP7A with accumulated platinum in question were fixed, dehydrated, infiltrated, and embedded with epoxy resin (Durcupan). The embedded material was sectioned at both 80um and 150um. Intitial EELS results from JEOL 3200EF were negative.