Anjali Bhat1, Aman Sharma2, Alok Chandra Bharti1. 1. Molecular Oncology Laboratory, Department of Zoology, North Campus, University Of Delhi, New Delhi 110007, India. 2. ExoCan Healthcare Technologies Pvt Ltd, Lab 4, 400 NCL Innovation Park, Dr Homi Bhabha Road, Pune 411008, India.
Abstract
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.
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.
Authors: Leandra B Jones; Sanjay Kumar; Aliyah J Curry; Jayde S Price; Alexandre Krendelchtchikov; Brennetta J Crenshaw; Courtnee' R Bell; Sparkle D Williams; Tambre A Tolliver; Sabita N Saldanha; Brian Sims; Qiana L Matthews Journal: Biomedicines Date: 2019-09-30