Hsa_circ_0060927 participates in the regulation of Caudatin on colorectal cancer malignant progression by sponging miR-421/miR-195-5p

Abstract

Background: Caudatin is extracted from radix cynanchi bungei and has an inhibitory effect on cancer progression. The study aims to reveal the impacts of hsa_circ_0060927 on Caudatin-mediated colorectal cancer (CRC) development and the underneath mechanism. Methods: The expression levels of hsa_circ_0060927, microRNA-421 (miR-421) and miR-195-5p were detected by quantitative real-time reverse transcription-polymerase chain reaction. The protein expression was analyzed by Western blot or immunohistochemistry assay. Cell viability and proliferation were analyzed by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide or 5-Ethynyl-29-deoxyuridine assay. Cell apoptosis was quantified by flow cytometry analysis. Cell migration and invasion were investigated by transwell assay. The putative associations among hsa_circ_0060927, miR-421 and miR-195-5p were predicted by the starbase online database, and identified by dual-luciferase reporter, RNA pull-down and RNA immunoprecipitation (RIP) assays. The impacts of Caudatin treatment on tumor growth in vivo were revealed by a xenograft tumor model assay. Results: Hsa_circ_0060927 expression was significantly upregulated, whereas miR-421 and miR-195-5p were downregulated in CRC tissues and cells compared with control groups. Hsa_circ_0060927 expression was closely associated with lymph node metastasis and tumor-node-metastasis stage. Caudatin treatment significantly decreased hsa_circ_0060927 expression but increased miR-421 and miR-195-5p expression. Caudatin exposure suppressed CRC cell proliferation, migration and invasion, and induced cell apoptosis; however, hsa_circ_0060927 overexpression hindered these impacts. Additionally, hsa_circ_0060927 was associated with miR-421/miR-195-5p. Depletion of miR-421 or miR-195-5p attenuated the influences of hsa_circ_0060927 silencing on CRC development. Furthermore, Caudatin treatment repressed tumor growth in vivo. Conclusion: Caudatin inhibited CRC cell malignancy through the hsa_circ_0060927/miR-421/miR-195-5p pathway, which provided a potential therapeutic agent for CRC.