ACCUMULATION OF PLATINUM IN VESICLES FROM ATP7A-EXPRESSING CELLS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。最近的研究表明，铜转运蛋白在顺铂的运输中的作用。一些铜转运蛋白（ATP 7A、ATP 7 B和CTR 1）的表达改变导致顺铂和其他含铂化疗药物的蓄积和敏感性改变。我们一直在研究铂的积累和敏感性，使用从缺乏ATP 7A表达的Menkes患者中获得的成纤维细胞Me 32a，以及分子工程改造过表达ATP 7A的相同细胞系MeMNK。我们目前的研究表明，与非表达Me 32a细胞相比，顺铂处理的MeMNK细胞中铂的全细胞和囊泡蓄积更高。虽然这些结果表明ATP 7A在铂积累中的作用，但我们希望有一种更直接的方法来确定含有ATP 7A的囊泡是否也是在处理的细胞中积累铂的囊泡。更新：将具有ATP 7A和所讨论的累积铂的囊泡固定、脱水、渗透并用环氧树脂（Durcupan）包埋。将包埋材料切成80 μ m和150 μ m两种切片。JEOL 3200 EF的初始EELS结果为阴性。

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.