ABSTRACT
Extracellular vesicles (EVs) are shed by many different cell types. Their nucleic acids content offers new opportunities for biomarker research in different solid tumors. The role of EV RNA in prostate cancer (PCa) is still largely unknown. EVs were isolated from different benign and malignant prostate cell lines and blood plasma from patients with PCa (n = 18) and controls with benign prostatic hyperplasia (BPH) (n = 7). Nanoparticle tracking analysis (NTA), Western blot, electron microscopy, and flow cytometry analysis were used for the characterization of EVs. Non-coding RNA expression profiling of PC3 metastatic PCa cells and their EVs was performed by next generation sequencing (NGS). miRNAs differentially expressed in PC3 EVs were validated with qRT-PCR in EVs derived from additional cell lines and patient plasma and from matched tissue samples. 92 miRNAs were enriched and 48 miRNAs were depleted in PC3 EVs compared to PC3 cells, which could be confirmed by qRT-PCR. miR-99b-5p was significantly higher expressed in malignant compared to benign EVs. Furthermore, expression profiling showed miR-10a-5p (p = 0.018) and miR-29b-3p (p = 0.002), but not miR-99b-5p, to be overexpressed in plasma-derived EVs from patients with PCa compared with controls. In the corresponding tissue samples, no significant differences in the miRNA expression could be observed. We thus propose that EV-associated miR-10a-5p and miR-29b-3p could serve as potential new PCa detection markers.
ABSTRACT
OBJECTIVE: Various pleiotropic substances have been suggested as candidates that directly reduce the severity of liver injury after hepatic ischemia/reperfusion (I/R) and upon acute liver failure (ALF). Herein, we studied whether thrombopoietin (TPO), the main regulator of megakaryopoiesis and thrombopoiesis, showed hepatoprotective effects and might mediate an antiapoptotic function in liver tissue under stress. METHODS/RESULTS: In livers with ALF or undergoing warm hepatic I/R, injury was quantified by intravital fluorescence microscopy, chemical, and immunohistochemical analysis as well as western immunoblot. Induction of both ALF and I/R injury led to hepatocellular expression of c-mpl, the receptor of TPO. Exogenous application of recombinant TPO in a low (12.5 microg/kg) as well as a high (75 microg/kg) dose, however, did not ameliorate postischemic perfusion and leukocyte endothelial cell interaction, but slightly aggravated transaminase release upon I/R. Similarly, TPO was unable to dampen hepatic microcirculatory deteriorations after the induction of ALF, but caused an increase of leukocyte accumulation and transaminase activity when applied in high dose. Low dose of TPO did not influence the amount of hepatocellular apoptosis, whereas high-dose TPO slightly diminished the activation of caspase 3. Interestingly, exogenous TPO application completely reversed the stress-induced increase of plasma IL-6 levels, suggesting a negative feedback of TPO on IL-6 release. CONCLUSION: Although the existence of the TPO-receptor on target liver cells TPO plays only a minor role in mediating hepatocyte apoptosis and does not provide protection against hepatic injury, contrasting the efficacy of the related hematopoietic growth factor erythropoietin.
