Upstream Hedgehog signaling components are exported in exosomes of cervical cancer cell lines

Aim: To investigate export of Hedgehog pathway (Hh) proteins Patched1, Smoothened, Sonic hedgehog and Indian hedgehog in cervical cancer cell line (CaCx) exosomes. Methods: Exosomes were isolated and characterized by Western blotting, scanning electron microscopy and in a colorimetric assay. Nucleic acids (RNA, DNA) and protein content of exosomes were analyzed. Hh pathway proteins in exosomes were detected using Western blotting. Results: CaCx secrete bio-macromolecule (DNA, RNA and proteins) enriched exosomes. CaCx exosomes contained higher amount of RNA with respect to DNA. CaCx preferentially exported Hh proteins (Patched1, Smoothened, Sonic hedgehog, Indian hedgehog) in their exosomes. Cellular uptake assay revealed rapid internalization of CaCx exosomes in human umbilical vein endothelial cells. Conclusion: Our study showed that Hh proteins are exported in CaCx exosomes.