Department of General Surgery, Cangzhou Central Hospital, CangZhou, Hebei Province, China.
Department of Urology, Cangzhou People's Hospital, CangZhou, Hebei Province, China.
Bioengineered. 2022 May;13(5):11810-11821. doi: 10.1080/21655979.2022.2073129.
This study aimed to investigate the molecular mechanism of circular RNA circ-0039459 and its effects on the apoptosis, proliferation, invasion, and migration of hepatocellular carcinoma cells. The expression of circ-0039459, miR-432, and synoviolin 1 (SYVN1) mRNA was determined using real-time quantitative reverse transcription PCR. Cell proliferation was detected by cell counting kit-8 assay, and the apoptosis rate was detected using flow cytometry. Cell migration and invasion were detected using Transwell assay. The expression of E-cadherin, N-cadherin, and vimentin was detected using western blot. The targeting relationship between circ-0039459 and miR-432 as well as that between miR-432 and SYVN1 were detected using the dual-luciferase reporter and RNA pull-down assays. We found that circ-0039459 and SYVN1 mRNA were highly expressed, whereas miR-432 was lowly expressed in hepatocellular carcinoma cells and tissues. After treatment with ribonuclease R or actinomycin D, the expression of linear RNA was reduced, whereas that of circular RNA was not significantly changed. circ-0039459 knockdown or miR-432 overexpression can inhibit cell proliferation, invasion, and migration and the expression of N-cadherin and vimentin proteins in carcinoma cells as well as promote apoptosis and increase the E-cadherin level. circ-0039459 targeted and regulated miR-432, which targeted and regulated SYVN1. The decreased miR-432 expression reversed the effects of circ-0039459 knockout in cancer cells. Furthermore, SYVN1 overexpression reversed the effect of miR-432 overexpression in hepatoma cells. Hence, circ-0039459 can affect the proliferation, apoptosis, migration, and invasion of hepatocellular carcinoma cells through the adsorption of miR-432, thereby regulating the expression of SYVN1.
本研究旨在探讨环状 RNA circ-0039459 的分子机制及其对肝癌细胞凋亡、增殖、侵袭和迁移的影响。采用实时定量逆转录 PCR 检测 circ-0039459、miR-432 和滑膜素 1 (SYVN1)mRNA 的表达。通过细胞计数试剂盒-8 检测细胞增殖,通过流式细胞术检测细胞凋亡率。通过 Transwell 检测细胞迁移和侵袭。采用 Western blot 检测 E-钙黏蛋白、N-钙黏蛋白和波形蛋白的表达。采用双荧光素酶报告和 RNA 下拉实验检测 circ-0039459 与 miR-432 以及 miR-432 与 SYVN1 的靶向关系。我们发现,circ-0039459 和 SYVN1mRNA 在肝癌细胞和组织中高表达,而 miR-432 低表达。用核糖核酸酶 R 或放线菌素 D 处理后,线性 RNA 的表达减少,而环状 RNA 的表达无明显变化。circ-0039459 敲低或 miR-432 过表达可抑制癌细胞增殖、侵袭和迁移及 N-钙黏蛋白和波形蛋白蛋白的表达,并促进细胞凋亡,增加 E-钙黏蛋白水平。circ-0039459 靶向并调节 miR-432,后者靶向并调节 SYVN1。miR-432 表达降低可逆转 circ-0039459 敲除在癌细胞中的作用。此外,SYVN1 过表达可逆转 miR-432 过表达对肝癌细胞的作用。因此,circ-0039459 可通过吸附 miR-432 影响肝癌细胞的增殖、凋亡、迁移和侵袭,从而调节 SYVN1 的表达。