Department of Hepatobiliary Surgery, Jining No.1 People's Hospital, Jining, Shandong, China.
Department of Gynecology, Jining Hospital of TCM, Jining, Shandong, China.
J Cell Biochem. 2019 Aug;120(8):13361-13371. doi: 10.1002/jcb.28611. Epub 2019 Mar 27.
Hepatocellular carcinoma (HCC) is the most major type of primary hepatic cancer. This study aimed to explore the possible oncogenic effects of the long noncoding RNA cardiac hypertrophy-related factor (CHRF) on HCC, as well as the underlying possible mechanism.
The expression levels of CHRF and microRNA-211 (miR-211) in HCC tissues and/or cell lines HepG2 and Huh-7 were measured using quantitative reverse transcription polymerase chain reaction. Cell transfection was conducted to change the expression levels of CHRF and miR-211 in cells. Cell viability and apoptosis were assessed using the cell counting kit-8 assay and annexin V-phycoerythrin staining, respectively. The pull-down assay and RNA immunoprecipitation were performed to analyze the association between CHRF and miR-211. The expression of the key factors involving in cell proliferation, cell apoptosis, and epithelial-mesenchymal transition (EMT) process, as well as the phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT) and Wnt/β-catenin pathways, were evaluated by Western blot analysis.
CHRF was highly expressed in HCC tissues and positively associated with the TNM stage, differentiation, and size of tumors. Overexpression of CHRF promoted HepG2 cell viability, proliferation, and EMT process. CHRF knockdown had opposite effects. Moreover, CHRF negatively regulated the expression of miR-21, and miR-21 was a direct target of CHRF. Overexpression of miR-211 reversed the effects of CHRF on HepG2 and Huh-7 cell viability, proliferation, and EMT process. Furthermore, overexpression of CHRF activated the PI3K/AKT and Wnt/β-catenin pathways in HepG2 cells by downregulating miR-211.
CHRF played oncogenic roles in HCC. The overexpression of CHRF promoted HepG2 and Huh-7 cell viability, proliferation, and EMT process by downregulating miR-211 and then activating the PI3K/AKT and Wnt/β-catenin pathways.
肝细胞癌（HCC）是原发性肝癌最主要的类型。本研究旨在探讨长链非编码 RNA 心脏肥大相关因子（CHRF）对 HCC 的致癌作用及其潜在机制。
采用实时定量逆转录聚合酶链反应检测 HCC 组织及 HepG2、Huh-7 细胞系中 CHRF 和 microRNA-211（miR-211）的表达水平。通过细胞转染改变细胞中 CHRF 和 miR-211 的表达水平。采用细胞计数试剂盒-8 检测细胞活力，采用 Annexin V-phycoerythrin 染色检测细胞凋亡。采用 pull-down 实验和 RNA 免疫沉淀分析 CHRF 与 miR-211 的相关性。采用 Western blot 分析检测细胞增殖、细胞凋亡、上皮-间充质转化（EMT）过程中关键因子以及磷酸肌醇 3-激酶（PI3K）/蛋白激酶 B（AKT）和 Wnt/β-连环蛋白通路的表达。
CHRF 在 HCC 组织中高表达，与 TNM 分期、分化和肿瘤大小呈正相关。过表达 CHRF 促进 HepG2 细胞活力、增殖和 EMT 过程。CHRF 敲低则产生相反的作用。此外，CHRF 负调控 miR-21 的表达，miR-21 是 CHRF 的直接靶点。过表达 miR-211 逆转了 CHRF 对 HepG2 和 Huh-7 细胞活力、增殖和 EMT 过程的影响。此外，过表达 CHRF 通过下调 miR-211 激活 HepG2 细胞中的 PI3K/AKT 和 Wnt/β-连环蛋白通路。
CHRF 在 HCC 中发挥致癌作用。CHRF 过表达通过下调 miR-211 促进 HepG2 和 Huh-7 细胞活力、增殖和 EMT 过程，进而激活 PI3K/AKT 和 Wnt/β-连环蛋白通路。
