miR-27a-3p attenuates induce autophagy-related cell death by suppressing Inhibitor of growth family 5 in cholangiocarcinoma

Abstract

Background MicroRNAs (miRNAs) are abnormally expressed in human tumors, including cholangiocarcinoma (CCA). miR-27a-3p was observed up-regulated in CCA, but its functions in CCA are largely unknown.Methods CCK8 assay, Colony formation assays and Ki-67 staining was employed to detect the cell growth. The autophagy and proliferation relative-protein analyzed by western blot. The immunofluorescence staining was applied to analyze the expression level of LC3 I/II. Tumor xenografts was used to test the role of miR-27a-3p. Luciferase reporter assay, western bolt and qRT-PCR showed the relationship between miR-27a-3p and ING5.Results miR-27a-3p expression was increased in human CCA tissues. Inhibition of miR-27a-3p suppressed the proliferative capacity of CCA cells, silencing of miR-27a-3p dramatically induced cell death and suppressed tumor growth in vivo. The proteins, such as Beclin-1, p62, p21, p-p53, CDK4 and CDK6, were decreased upon miR-27a-3p inhibitor transfection. Western blot assay and immunofluorescence analysis were showed the induced-autophagy after transfecting with miR-27a-3p or inhibitor of growth family 5 (ING5) in RBE. ING5 as a direct miR-27a-3p target in CCA. Co-transfect of miR-27a-3p and ING5 can reverse CCA cell death which induced by miR-27a-3p inhibitor alone.Conclusions miR-27a-3p promotes oncogenesis of CCA by triggering autophagy-related cell death by interacting with ING5 directly.