PRP?1 significantly decreases the ALDHhigh cancer stem cell population and regulates the aberrant Wnt/??catenin pathway in human chondrosarcoma JJ012 cells.

ABSTRACT: Chondrosarcomas are malignant bone tumors refractory to chemotherapy and radiation treatment; thus, novel therapeutic strategies are required. Proline?rich polypeptide 1 (PRP?1) has previously demonstrated antitumor properties in chondrosarcoma. To further investigate the role of PRP?1 in chondrosarcoma cells, its effects on cancer stem cell (CSC) populations were determined by analyzing aldehyde dehydrogenase (ALDH) activity, an established marker of CSCs, in association with regulation of the Wnt/??catenin signaling. A significant decrease in ALDHhigh CSCs was observed following treatment of chondrosarcoma JJ012 cells with PRP?1. For RT2 profiler PCR array analysis of Wnt/??catenin signaling genes, cells were sorted into: i) Bulk JJ012 cells; ii) ALDHhigh cells sorted from untreated JJ012 cells (ALDHhigh?untreated); and iii) ALDHlow cells sorted from PRP?1?treated JJ012 cells (ALDHlow?PRP?1). The expression levels of Wnt/??catenin signaling genes were determined to be downregulated in the ALDHhigh?untreated cells and upregulated in ALDHlow?PRP?1 cells when compared to the bulk JJ012 cells. Additionally, two important oncogenes involved in this pathway, MMP7 and CCND2, were found to be downregulated in the ALDHlow?PRP?1 cells. Immunocytochemistry demonstrated the localization of ??catenin in the nuclei of the PRP?1?treated cells. Western blotting indicated increased ??catenin expression in the ALDHlow?PRP?1 cells compared with the bulk JJ012 cells. Analysis of the cytoplasmic and nuclear fractions of cells treated with increasing concentrations of PRP?1 and ??catenin nuclear translocation inhibitor CGP57380, suggested the nuclear translocation of ??catenin following PRP?1 treatment. In addition, treatment of JJ012 cells with a specific ALDH inhibitor, diethylaminobenzaldehyde, and PRP?1 resulted in a significant decrease in cytoplasmic ??catenin protein expression. This indicated that ALDH inactivation may be associated with the nuclear translocation of ??catenin. Derivation of sarcomas from mesenchymal stem cells via inactivation of the Wnt pathway has been previously documented. The findings of the present study support the notion that Wnt/??catenin activation may serve a differential role in sarcomas, limiting tumor progression in association with decreased CSC activity.

SUBMITTER: Hoyt AK 

PROVIDER: S-EPMC6549102 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature