Hubei Key Laboratory of Tumor Biological Behaviors, Zhongnan Hospital of Wuhan University, Wuhan, 430071, People's Republic of China.
Department of Radiation and Medical Oncology, Zhongnan Hospital of Wuhan University, Wuhan, 430071, People's Republic of China.
Clin Transl Oncol. 2021 Mar;23(3):468-480. doi: 10.1007/s12094-020-02440-5. Epub 2020 Jul 2.
To explore FGF1 and miR-143-3p expression in hepatocellular carcinoma (HCC) cells and its related mechanisms.
Eighty-two HCC patients treated at our hospital from January 2018 to January 2019 were enrolled as Group A, while further 80 healthy people undergoing physical examinations during the same time period were enrolled as Group B. HCC cells and normal human liver cells were purchased, with HepG2 and SMMC-7721 cells transfected with pcDNA3.1-FGF1, si-FGF1, NC, miR-143-3p-inhibitor and miR-143-3p-mimics. FGF1 and miR-143-3p expression was detected by qRT-PCR. The expression of N-cadherin, vimentin, Snail, Slug, E-cadherin and γ-catenin was detected by Western Blotting (WB). Cell proliferation was detected by MTT assay. Cell invasion was detected by Transwell. Cell apoptosis was detected by flow cytometry (FCM).
FGF1 was highly expressed but miR-143-3p was poorly expressed in HCC cells. Areas under the curves (AUCs) of the two indicators were > 0.8. The indicators were correlated with the age, gender, tumor invasion, degree of differentiation, tumor location and TNM staging of the patients. Silencing FGF1 and overexpressing miR-143-3p could promote cell apoptosis, inhibit cell growth, cell epithelial-mesenchymal transition (EMT) and the expression of N-cadherin, vimentin, Snail and Slug, and increase the expression of E-cadherin and γ-catenin. Dual luciferase reporter gene assay (DLRGA) confirmed that FGF1 and miR-143-3p had a targeted relationship. The rescue experiment showed that the proliferation, invasion and apoptosis of HepG2 and SMMC-7721 cells in the miR-143-3p-mimics+pcDNA3.1-FGF1 and miR-143-3p-inhibitor+Si-FGF1 groups were not different from those in the miR-NC group.
Inhibiting FGF1 can upregulate miR-143-3p-mediated Hedgehog signaling pathway, and affect cells' EMT, proliferation and invasion, so FGF1 is expected to become a potential therapeutic target for HCC.
探讨成纤维细胞生长因子 1（FGF1）和微小 RNA-143-3p（miR-143-3p）在肝细胞癌（HCC）细胞中的表达及其相关机制。
选取 2018 年 1 月至 2019 年 1 月我院收治的 82 例 HCC 患者为 A 组，另选取同期体检的 80 例健康人为 B 组。购买 HCC 细胞和正常人类肝细胞，用 pcDNA3.1-FGF1、si-FGF1、NC、miR-143-3p 抑制剂和 miR-143-3p 模拟物转染 HepG2 和 SMMC-7721 细胞。用 qRT-PCR 检测 FGF1 和 miR-143-3p 的表达。用 Western Blotting（WB）检测 N-钙黏蛋白、波形蛋白、Snail、Slug、E-钙黏蛋白和 γ-连环蛋白的表达。用 MTT 检测细胞增殖。用 Transwell 检测细胞侵袭。用流式细胞术（FCM）检测细胞凋亡。
HCC 细胞中 FGF1 高表达而 miR-143-3p 低表达。两个指标的曲线下面积（AUC）均>0.8。这些指标与患者的年龄、性别、肿瘤侵袭、分化程度、肿瘤位置和 TNM 分期有关。沉默 FGF1 和过表达 miR-143-3p 可促进细胞凋亡，抑制细胞生长、上皮-间质转化（EMT）以及 N-钙黏蛋白、波形蛋白、Snail 和 Slug 的表达，增加 E-钙黏蛋白和 γ-连环蛋白的表达。双荧光素酶报告基因检测（DLRGA）证实 FGF1 和 miR-143-3p 存在靶向关系。挽救实验表明，miR-143-3p 模拟物+pcDNA3.1-FGF1 和 miR-143-3p 抑制剂+Si-FGF1 组 HepG2 和 SMMC-7721 细胞的增殖、侵袭和凋亡与 miR-NC 组无差异。
抑制 FGF1 可上调 miR-143-3p 介导的 Hedgehog 信号通路，影响细胞 EMT、增殖和侵袭，因此 FGF1 有望成为 HCC 的潜在治疗靶点。
