ACCUMULATION OF PLATINUM IN VESICLES FROM ATP7A-EXPRESSING CELLS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。最近的研究表明，铜转运体在顺铂的运输中发挥了作用。一些铜转运蛋白(ATP7A、ATP7B和CTR1)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ATP7A表达的Menkes患者的成纤维细胞Me32a，以及相同的细胞系分子工程过表达ATP7A，MeMNK。我们目前的研究表明，与不表达Me32a的细胞相比，经顺铂处理的MeMNK细胞中铂的整体和囊泡积累更高。虽然这些结果表明ATP7A在铂的积累中发挥了作用，但我们希望有一种更直接的方法来确定含有ATP7A的囊泡是否也是处理细胞中积累铂的囊泡。更新：含有ATP7A的囊泡是固定的，脱水，渗透，并用环氧树脂(杜卡潘)包埋。包埋材料分别在80um和150um处切片。JEOL 3200EF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.