Department of Neurology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China.
Eur Rev Med Pharmacol Sci. 2020 Dec;24(23):12144-12152. doi: 10.26355/eurrev_202012_24003.
The purpose of this study was to investigate the expression level of microRNA-30a-3p in hepatocellular carcinoma (HCC), and to further study its relationship with HCC clinical parameters and prognosis and the underlying mechanisms.
Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine microRNA-30a-3p level in 44 tumor tissue specimens and paracancerous normal ones collected from HCC patients, and the interplay between microRNA-30a-3p expression and clinical indicators, as well as prognosis of HCC patients was analyzed. Meanwhile, qPCR was also used to further verify microRNA-30a-3p expression in HCC cell lines. In addition, microRNA-30a-3p overexpression and knockdown models were constructed in HCC cell lines, and the impacts of microRNA-30a-3p on HCC cell functions was evaluated by cell counting kit-8 (CCK-8), transwell and cell wound healing assays. Finally, the Luciferase reporting assay was conducted to uncover the underlying mechanism.
In this study, qRT-PCR results showed that the expression level of microRNA-30a-3p in tumor tissues of HCC patients was markedly lower than that in adjacent ones. Compared with patients with high expression of microRNA-30a-3p, the patients with low expression of microRNA-30a-3p had a higher incidence of lymphatic or distant metastasis and a lower overall survival rate. In the Bel-7402 cell line, the proliferation, invasion, and metastasis ability of HCC cells were decreased markedly after microRNA-30a-3p overexpression, while in Hep3B cell line, knockdown of microRNA-30a-3p enhanced the cell proliferation and invasion capacity. In addition, Luciferase reporting assay demonstrated that microRNA-30a-3p could specifically bind to IGF1. Furthermore, Western Blot results also verified a reduced expression of IGF1 after overexpression of microRNA-30a-3p, and an elevated one after knockdown of microRNA-30a-3p. Finally, cell recovery experiment verified that microRNA-30a-3p and IGF1 may regulate each other and thereby together inhibit the malignant progression of HCC.
MicroRNA-30a-3p expression is significantly decreased in HCC tumor tissue samples, which is associated with lymph node or distant metastasis rate, as well as the poor prognosis of HCC. In addition, this research suggests that microRNA-30a-3p may inhibit the malignant progression of HCC by regulating IGF1.
本研究旨在探讨 microRNA-30a-3p 在肝细胞癌（HCC）中的表达水平，并进一步研究其与 HCC 临床参数和预后的关系及其潜在机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 44 例 HCC 患者肿瘤组织标本和癌旁正常组织中 microRNA-30a-3p 的水平，分析 microRNA-30a-3p 表达与 HCC 患者临床指标及预后的关系。同时，qPCR 进一步验证了 HCC 细胞系中 microRNA-30a-3p 的表达。此外，构建 HCC 细胞系中 microRNA-30a-3p 的过表达和敲低模型，通过细胞计数试剂盒-8（CCK-8）、Transwell 和细胞划痕愈合实验评估 microRNA-30a-3p 对 HCC 细胞功能的影响。最后，通过荧光素酶报告基因检测揭示其潜在机制。
本研究中，qRT-PCR 结果显示 HCC 患者肿瘤组织中 microRNA-30a-3p 的表达水平明显低于癌旁组织。与 microRNA-30a-3p 高表达患者相比，microRNA-30a-3p 低表达患者的淋巴或远处转移发生率更高，总生存率更低。在 Bel-7402 细胞系中，microRNA-30a-3p 过表达后 HCC 细胞的增殖、侵袭和转移能力明显下降，而在 Hep3B 细胞系中，microRNA-30a-3p 敲低增强了细胞的增殖和侵袭能力。此外，荧光素酶报告基因检测表明 microRNA-30a-3p 可以特异性结合 IGF1。Western blot 结果也验证了 microRNA-30a-3p 过表达后 IGF1 的表达下调，而 microRNA-30a-3p 敲低后 IGF1 的表达上调。最后，细胞恢复实验验证了 microRNA-30a-3p 和 IGF1 可能相互调节，从而共同抑制 HCC 的恶性进展。
microRNA-30a-3p 在 HCC 肿瘤组织样本中的表达显著降低，与淋巴结或远处转移率以及 HCC 的不良预后相关。此外，本研究表明，microRNA-30a-3p 可能通过调节 IGF1 抑制 HCC 的恶性进展。
