GSE196131
Extracellular vesicle-mediated RNA transfer contributes to inter-cellular communication in the liver tumor microenvironment
Extracellular vesicles and their contents are gaining recognition as important mediators of intercellular communication through the transfer of bioactive molecules such as non-coding RNA. We evaluated the contribution of EV miRNA in intercellular communication between hepatocellular cancer cells and hepatic stellate cells. EV-based miRNA was comprehensively assessed in both cell types individually. Using co-cultures of both HCC and HSC cells in transwell and 3D spheroids culture, we established EV-based miR-126-3p as a potential EV-based miRNA mediator of HSC to HCC communication through which tumor cell migration, invasion and three-dimensional growth in spheroids could be influenced. Manipulation of miR-126-3p by enforced expression or inhibition using pre-miR-126-3p or antimiR-126-3p respectively did not alter cell viability, proliferation, or sensitivity to either sorafenib or regorafenib. In contrast, migration was decreased in HepG2 cells with enforced expression of miR-126-3p. Knockdown of miR-126-3p in HepG2 resulted in a significant increase in ADAM9 expression and in LX-2 cells increased collagen accumulation and the compactness of spheroids but did not alter the number or size of spheroids. The restoration of miR-126 in 3D-co-culture of HepG2 and LX2 cells with precursor showed significant alleviated expression of ADAM9 and VEGF, While the silencing of miR-126 was elevated the expression of ADAM9 and VEGF. These studies implicate miR-126-3p in functional effects on migration, invasion, and spheroid growth in HCC cells in the presence of HSC, and thus demonstrate the presence of functional EV RNA based intercellular signaling between HSC and HCC cells that may be relevant to tumor cell behavior. Overall design: To evaluate functional role of extracelluar vesicle RNA contribution in the liver tumor microenvironment and to identify extracelluar vesicle RNA as biomarkers of tumor microenvironment.