Circ_0064288 通过抑制 miR-335-5p 的表达和促进 ROCK1 的表达，充当肝癌细胞的癌基因。
Circ_0064288 acts as an oncogene of hepatocellular carcinoma cells by inhibiting miR-335-5p expression and promoting ROCK1 expression.
机构信息
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.
出版信息
BMC Cancer. 2022 Mar 14;22(1):265. doi: 10.1186/s12885-022-09323-8.
BACKGROUND
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.
METHODS
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.
RESULTS
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.
CONCLUSION
Circ_0064288 facilitates HCC cell growth and migration by modulating the miR-335-5p/ROCK1 axis.
背景
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。
方法
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。
结果
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。
结论
circ_0064288 通过调节 miR-335-5p/ROCK1 轴促进 HCC 细胞的生长和迁移。
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