Department of Gastroenterology, Jiamusi Central Hospital, Jiamusi, 154002, Heilongjiang, China.
Department of Gastroenterology, the First Affiliated Hospital of Harbin Medical University, Harbin, 150000, Heilongjiang, China.
BMC Cancer. 2022 Mar 14;22(1):265. doi: 10.1186/s12885-022-09323-8.
Reportedly, circular RNA (circRNA) is a key modulator in the development of human malignancies. This work is aimed to probe the expression pattern, biological effects and mechanism of circ_0064288 on hepatocellular carcinoma (HCC) progression.
The differentially expressed circRNA was screened by analyzing the expression profiles of circRNAs in HCC tissues and normal tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to examine the expression of circ_0064288, miR-335-5p and Rho associated coiled-coil containing protein kinase 1 (ROCK1) mRNA in HCC specimens. After circ_0064288 was overexpressed or knocked down in HCC cells, cell growth was detected by the CCK-8 experiment, and cell migration was evaluated using Transwell experiment and scratch healing experiment. The targeting relationship between miR-335-5p and circ_0064288 and ROCK1 mRNA was predicted and verified using bioinformatic analysis and dual-luciferase reporter gene experiments, respectively. Western blot was executed to examine ROCK1 protein expression in HCC cells.
Circ_0064288 and ROCK1 expression was up-modulated in HCC, while miR-335-5p was down-modulated. High circ_0064288 expression was associated with shorter survival time of HCC patients. It was also revealed that circ_0064288 overexpression remarkably enhanced HCC cell growth and migration, while knockdown of circ_0064288 induced opposite effects. Additionally, circ_0064288 could competitively bind with miR-335-5p thereby up-modulate ROCK1 expression. MiR-335-5p overexpression partly counteracted the effect of circ_0064288 overexpression on HCC cells.
Circ_0064288 facilitates HCC cell growth and migration by modulating the miR-335-5p/ROCK1 axis.
据报道，环状 RNA（circRNA）是人类恶性肿瘤发展的关键调节剂。本研究旨在探讨 circ_0064288 对肝细胞癌（HCC）进展的表达模式、生物学效应和机制。
通过分析 HCC 组织和正常组织中 circRNAs 的表达谱筛选差异表达的 circRNA。采用定量实时聚合酶链反应（qRT-PCR）检测 HCC 标本中 circ_0064288、miR-335-5p 和 Rho 相关卷曲螺旋蛋白激酶 1（ROCK1）mRNA 的表达。在 HCC 细胞中转染 circ_0064288 过表达或敲低后，通过 CCK-8 实验检测细胞生长，通过 Transwell 实验和划痕愈合实验评估细胞迁移。通过生物信息学分析和双荧光素酶报告基因实验分别预测和验证 miR-335-5p 与 circ_0064288 和 ROCK1 mRNA 的靶向关系。Western blot 检测 HCC 细胞中 ROCK1 蛋白的表达。
circ_0064288 和 ROCK1 在 HCC 中表达上调，而 miR-335-5p 表达下调。高 circ_0064288 表达与 HCC 患者的生存时间缩短有关。研究还表明，circ_0064288 过表达显著增强了 HCC 细胞的生长和迁移，而 circ_0064288 敲低则诱导了相反的效果。此外，circ_0064288 可以与 miR-335-5p 竞争性结合，从而上调 ROCK1 的表达。miR-335-5p 过表达部分抵消了 circ_0064288 过表达对 HCC 细胞的影响。
circ_0064288 通过调节 miR-335-5p/ROCK1 轴促进 HCC 细胞的生长和迁移。
