Abstract 114P
Background
Exosomes (EXO) in cholangiocarcinoma (CCCs) are distinctly strong intermediator with ability for flipping the action of neighbouring cells. This becomes more obvious that exosomes have aptitude to encourage the development of pre metastatic niche. As for metastasis to happen, cholangiocarcinoma cells need to migrate in a new environment which also requires being favourable for tumor to colonize properly. Their release into the circulation has the potential to inform about tumor status. In-depth proteomic characterization of plasma-derived EXOs has been limited by challenges in isolating EXO s from protein-abundant biological fluids. We implemented a novel single-step density gradient flotation workflow for efficient and rapid isolation of highly enriched circulating EXOs from plasma-derived exosomes. Effective methods for EXOs are still in the waiting and there is strong need for high confidence identification of blood-based biomarkers. Exosomes have recently emerged as a novel source of circulatory biomarkers for cancer.
Methods
NTA and TEM confirmed the presence of plasma-derived exosomes (EXOs) isolated by differential centrifugation. Total proteins extracted from plasma-derived EXOs of 40 Cholangiocarcinoma and 40 healthy subjects was analysed. FACS analysis to characterized exosomal proteins tetraspanins CD63, CD9 and CD81 was demonstrated to be significantly increased in plasma-derived exosomes of Cholangiocarcinoma patients with controls. Western blot analysis of blood plasma-derived exosomes was carried out to identify proteins showing altered levels in Cholangiocarcinoma cases in comparison to controls. The result shows that CD63, CD9 and CD81 were detected in plasma-derived exosomes in Cholangiocarcinoma patients and healthy individuals.
Results
Proteomic analysis of blood plasma-derived exosomes was carried out to identify EXOs proteins showing altered levels in cases as comparison to healthy individuals.
Conclusions
Our findings support the potential of exosomes derived proteins as a source of biomarkers that complement other approaches for cancer assessment with the help of computational analysis of protein-protein interactions and their role in disease pathogenesis.
Legal entity responsible for the study
R. Rana.
Funding
Has not received any funding.
Disclosure
R. Rana: Financial Interests, Institutional, Invited Speaker: Sir Ganga Ram Hospital.