Exosome-mediated circTTLL5 transfer promotes hepatocellular carcinoma malignant progression through miR-136-5p/KIAA1522 axis.

Abstract

Exosomes have been recognized as messengers for intercellular communication in tumor microenvironment. Exosomal circRNAs are reported to be important in tumors. Here, this study identified the potential function of exosomal circular RNA tubulin tyrosine ligase like 5 (circTTLL5) in hepatocellular carcinoma (HCC) progression. The expression of circTTLL5, microRNA (miR)-136-5p and KIAA1522 was detected using qRT-PCR and Western blot assays. Exosomes were isolated by ultracentrifugation, and qualified by transmission electron microscopy (TEM) and Western blot. Cell proliferation, apoptosis and metastasis were investigated using cell counting kit-8, colony formation, flow cytometry, would healing, transwell and Western blot assays, respectively. The interaction between miR-136-5p and circTTLL5 or KIAA1522 was confirmed by dual-luciferase reporter and pull-down assays. In vivo experiment was performed using Xenograft models. CircTTLL5 was incorporated into exosomes and highly expressed in HCC tissues and cells. CircTTLL5 knockdown suppressed HCC cell proliferation and metastasis in vitro and impeded tumor growth in mice. CircTTLL5 could be delivered to recipient cells via exosomes, and treatment of circTTLL5-elevated exosomes could attenuate the anticancer effects of circTTLL5 knockdown on HCC in vitro and in vivo. Mechanically, circTTLL5 could sponge miR-136-5p, which controlled its down-stream target KIAA1522. MiR-136-5p inhibition reversed the effects of circTTLL5 knockdown on HCC cells. Besides that, miR-136-5p re-expression inhibited HCC cell growth and metastasis, which was abated by KIAA1522 overexpression. Exosomal circTTLL5 promoted HCC progression through miR-136-5p/KIAA1522 axis, suggesting that blockage of the exosome-mediated transfer of circTTLL5 might be a therapeutic target for HCC.