School of Nursing, Jiangsu Vocational College of Medicine, Yancheng, China.
Eur Rev Med Pharmacol Sci. 2019 Sep;23(18):7874-7883. doi: 10.26355/eurrev_201909_18997.
The aim of this study was to figure out the effect of microRNA-217 on the proliferation of hepatocellular carcinoma (HCC) cells, and to explore its influence on KLF5 expression and the underlying regulatory mechanisms.
Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of microRNA-217 in tumor tissues and paracancerous tissues of 60 patients with HCC. Meanwhile, the relationship between microRNA-217 expression and HCC pathological parameters was analyzed. In HCC cell lines, including HepG2 and Bel-7402, negative control group (NC) and microRNA-217 overexpression group were set up, and qRT-PCR was performed to further verify their transfection efficiency. In addition, Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU) assay were performed to analyze the effect of microRNA-217 on the biological function of HCC cells. Finally, the potential mechanism of KLF5, the downstream gene of microRNA-217, was explored using bioinformatics analysis and cell recovery experiments.
QRT-PCR results showed that microRNA-217 level in tumor tissues of HCC patients was conspicuously lower than that in adjacent tissues, and the difference was statistically significant. Compared with patients with high expression of microRNA-217, the pathological stage was higher and the overall survival rate was lower in patients with low expression. Compared with the NC group, the cell proliferation ability of the microRNA-217 mimics group was conspicuously decreased. Subsequently, in the HCC cell line and tissue verification, the expression of KLF5 was found remarkably increased, and microRNA-217 exhibited a negative correlation with KLF5 level. In addition, the overexpression of microRNA-217 conspicuously reduced the protein expression of CD31, Ki-67, c-Myc, MMP-2, and MMP-9. In cell recovery experiment, it was found that the overexpression of KLF5 could counteract the effect of microRNA-217 mimics on the cell proliferation of HCC, thereby inhibiting the malignant progression of this disease.
The above studies demonstrated that microRNA-217 was markedly associated with the pathological stage and poor prognosis of HCC, and could inhibit the malignant progression of this disease. In addition, our investigation has showed that microRNA-217 might be capable of inhibiting cell proliferation of HCC via regulating KLF5.
本研究旨在探讨 microRNA-217 对肝癌（HCC）细胞增殖的影响，并探讨其对 KLF5 表达的影响及其潜在的调控机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 60 例 HCC 患者肿瘤组织和癌旁组织中 microRNA-217 的表达情况，并分析其与 HCC 病理参数的关系。在 HepG2 和 Bel-7402 肝癌细胞系中，分别设立阴性对照组（NC）和 microRNA-217 过表达组，通过 qRT-PCR 进一步验证其转染效率。此外，采用细胞计数试剂盒-8（CCK-8）和 5-乙炔基-2'-脱氧尿苷（EdU）检测分析 microRNA-217 对 HCC 细胞生物学功能的影响。最后，通过生物信息学分析和细胞恢复实验探讨 microRNA-217 下游基因 KLF5 的潜在作用机制。
qRT-PCR 结果显示，HCC 患者肿瘤组织中 microRNA-217 的水平明显低于癌旁组织，差异具有统计学意义。与 microRNA-217 高表达患者相比，低表达患者的病理分期较高，总生存率较低。与 NC 组相比，microRNA-217 模拟物组的细胞增殖能力明显降低。随后，在 HCC 细胞系和组织验证中发现，KLF5 的表达明显增加，microRNA-217 与 KLF5 水平呈负相关。此外，microRNA-217 的过表达明显降低了 CD31、Ki-67、c-Myc、MMP-2 和 MMP-9 的蛋白表达。在细胞恢复实验中发现，KLF5 的过表达可以抵消 microRNA-217 模拟物对 HCC 细胞增殖的影响，从而抑制该疾病的恶性进展。
上述研究表明，microRNA-217 与 HCC 的病理分期和预后不良显著相关，可抑制该疾病的恶性进展。此外，本研究表明，microRNA-217 可能通过调节 KLF5 抑制 HCC 细胞的增殖。
