Temozolomide increases heat shock proteins in extracellular vesicles released from glioblastoma cells

Abstract

Background: Glioblastoma (GBM) is the most malignant and the fastest-progressing type of primary brain tumours. Temozolomide (TMZ) is a chemotherapeutic drug for the treatment of GBM. Extracellular vesicles (EVs) have been recently con rmed to have a substantial role in the GBM, and their contents released from GBM cells have been considered a target for treatment. The purpose of this study is to evaluate the impact of TMZ on heat shock proteins (HSPs) derived from EVs originated from GBM cell lines (U87-MG and LN229) and the signi cance of EVs in response to chemotherapy in GBM.Methods and Results: NTA, ELISA, and immunoblotting were used to characterization studies of EVs and results showed that U87-MG cells released many EVs compared to LN229 cells. The effect of TMZ treatments on HSPs expression levels were assessed with immunoblotting and was found to be led to increases in HSF-1, Hsp90, Hsp70, Hsp60 and Hsp27 expression in GBM cells and their EV contents, which these increases are related to therapeutic resistance. What is more, in Real-time PCR studies showing which signalling pathways might be associated with these increases, it was observed that TMZ triggered the expression of RAD51 and MDM2 genes in cells and EV contents. More strikingly, we discover a correlation between EV and parental cells in regard of mRNA and protein level in both cell lines as a result of TMZ treatment. Conclusions: Our data suggest of EVs in the treatment of GBM may have potential biomarkers that can be used to investigate the treatment response.

Sequential Treatment with Temozolomide Plus Naturally Derived AT101 as an Alternative Therapeutic Strategy: Insights into Chemoresistance Mechanisms of Surviving Glioblastoma Cells

“…Kıyga et al [78] showed that the amount of CD63 in the EVs of TMZ (200 µM)treated GBM cells increased, however, without any statistically significant difference. Ricklefs et al [79] demonstrated that EVs with double positive tetraspanin expression (CD63 + /CD81 + ; CD9 + /CD81 + ) were enriched in GBM cell lines.…”

Extracellular vesicles as tools and targets in therapy for diseases

“… CircBACH1/ miR-217/G3BP2 axis a new regulatory for PTX-resistance and progression of breast cancer. 672 Cisplatin Ovarian cancer cells A2780, IGROV-1 cells Ovarian cancer CREB, ERK, JNK, p38α, p53 inhibits ovarian cancer 673 , 674 Milk exosomes loaded with cisplatin A2780, nu/nu mice Ovarian cancer ARF6, Rac1, CLTC, caveolin Overcomes cisplatin-resistance in ovarian cancer Temozolomide Glioblastoma cells U87-MG cells, GBM cells Glioblastoma Heat shock protein, RAD51, MDM2 Leads to cell adhesion 675 EVs encapsulated miR-153-3p LUAD cells NCI-H1993, SW1271, BALB/c mice, Lung adenocarcinoma BANCR, miR-153-3p, PI3K/AKT Increases cell invasion 676 EVs PDAC cells MIA PaCa-2, PANC-1,Rag2 −/− Ilrg2 −/−, mice Pancreatic ductal adenocarcinoma Rab27a, LRP-4 receptor, YAP Intratumor communication Targeted therapy for PDAC 677 EVs Human Liver Stem Cells MSCs Renal cancer cells SCID mice Renal cell carcinoma miR-Let7b, miR-200b, miR-200c and miR-223, EGFR, ZEB1, ZEB2, MMP1 antitumor effects 678 …”

Heat Shock Proteins Mediate Intercellular Communications within the Tumor Microenvironment through Extracellular Vesicles

“…In a combined TMZ plus an HSP90 inhibitor (geldanalmycin) treatment, there were more glioblastoma cells undergoing apoptosis and diminished vesiculation [72]. Kıyga et al observed increased expression of HSP70, HSP60, HSP90, and HSP27 (HSPB family) on EVs derived from glioblastoma cells after treatment and this increased expression was related to therapeutic resistance [73]. Lv et al showed that chemotherapeutic treatment of hepatocarcinoma cells, HepG2 and PLC/PRF/5, increases secretion of exosomes expressing HSP60, HSP70, and HSP90 on their membranes.…”