RESUMO
Paracrine and long-range signaling via extracellular vesicles, such as exosomes and microvesicles, is deemed crucial for tumorigenesis, invasion and spread of solid tumors. The ESCRT machinery (endosomal sorting complexes required for transport) and Rab-proteins act as key players in vesicular trafficking and secretion. Yet, their role in prostate cancer (PCa) is unknown. Therefore, this study aimed to elucidate the relevance of these components in PCa. In silico reanalysis of genes with known involvement in vesicular trafficking and secretion in an existing microarray dataset revealed low expression of RAB27A, RAB27B and VPS36 to be predictive for reduced BCR-free survival in patients with localized PCa (p=0.033, 0.025 and 0.005). In the same microarray dataset underexpression of RAB27A, RAB27B and VPS36 was seen in distant metastases (p<0.001; p=0.003; p<0.001). This was consistent in two further microarray datasets. qRT-PCR-validation in two independent cohorts of PCa specimens (n=90) showed low expression of VPS36 in PCa tissue (p=0.023), especially in castration-resistant tumors (p=0.002). In all five datasets there were significant correlations between the expression of at least two of the candidates. Upon knockdown of VPS36 an increase of RAB27A and RAB27B expression, but not vice versa, was observed in both prostate and breast cancer cells (PC3, MDA-MB231). In PC3 cell knockdown of RAB27B and VPS36 dramatically reduced colony formation (-52.2%, p<0.001; -71.1%, p<0.001) and, controversial to reports in other tumor entities, increased the release of extracellular particles (+25.3%, p=0.014; +45.6%, p<0.001). Taken together RAB27A, RAB27B and VPS36 are frequently underexpressed in advanced PCa and are inversely correlated with PCa outcome. There seems to be a close relationship in the expression of RAB27A, RAB27B and VPS36, with RAB27A and RAB27B being dependent on VPS36. Changes in colony formation and particle release upon RNAi indicate an involvement in paracrine cell-cell communication.