Zhang Qianqian, Deng Xiaohong, Tang Xiuxin, You Ying, Mei Meihua, Liu Danping, Gui Lian, Cai Yan, Xin Xiaoping, He Xiaoshun, Huang Junqi
Guangdong Provincial Key Laboratory of Biomedical Imaging and Guangdong Provincial Engineering Research Center of Molecular Imaging, The Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai, China.
Organ Transplant Center, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Front Oncol. 2021 Dec 16;11:737986. doi: 10.3389/fonc.2021.737986. eCollection 2021.
Hepatocellular carcinoma (HCC), a worldwide leading cause of morbidity and mortality, is the most frequent primary liver tumor. Most HCC patients are diagnosed with advanced liver cancer, resulting in a very low 5-year survival rate. Thus, there is an urgent need for the development of targeted therapies. In this study, we aimed to investigate the effect and mechanism of the miR-20a/EZH1 axis on the proliferation and metastasis of HCC and the inhibitory effect of the EZH1/EZH2 inhibitor UNC1999 on HCC.
The expression of miR-20a in human HCC tissues and cell lines was detected using quantitative real-time PCR (qRT-PCR). The expressions of proteins were analyzed with immunohistochemistry and Western blotting. Luciferase assay was used to verify whether miR-20a targets EZH1 or EZH2. The effect of miR-20a on HCC progression was studied and . The tumor inhibitory effect of UNC1999 was confirmed in vivo. CCK8 assay, wound healing assay, cell migration and invasion assay were used to evaluate the synergistic effect of UNC1999 with sorafenib. RNA sequencing (RNA-seq) was performed to screen the differentially expressed genes in the Huh7 and SMMC7721 cell lines after UNC1999, sorafenib, and combination treatments.
In this study, miR-20a showed a lower expression in both HCC tissues and cell lines. MiR-20a inhibited the proliferation and migration of SMMC7721 and Huh7 cells. The results of the luciferase assay and Western blot analysis revealed that miR-20a directly targeted EZH1, a histone methyltransferase. We demonstrated that miR-20a negatively regulated the expression of EZH1 and inhibited the proliferation and metastasis of HCC by reducing H3K27 methylation. We found UNC1999 inhibited tumor cells proliferation and enhanced the inhibitory effect of sorafenib.
We demonstrated that miR-20a suppresses the tumor proliferation and metastasis in HCC by directly targeting EZH1. UNC1999 can inhibit tumor proliferation and increase the sensitivity of hepatoma cell lines to sorafenib.
肝细胞癌（HCC）是全球发病率和死亡率的主要原因，是最常见的原发性肝脏肿瘤。大多数HCC患者被诊断为晚期肝癌，导致5年生存率极低。因此，迫切需要开发靶向治疗方法。在本研究中，我们旨在研究miR-20a/EZH1轴对HCC增殖和转移的影响以及EZH1/EZH2抑制剂UNC1999对HCC的抑制作用。
采用定量实时PCR（qRT-PCR）检测人HCC组织和细胞系中miR-20a的表达。用免疫组织化学和蛋白质印迹法分析蛋白质表达。荧光素酶报告基因检测用于验证miR-20a是否靶向EZH1或EZH2。研究了miR-20a对HCC进展的影响。在体内证实了UNC1999的肿瘤抑制作用。采用CCK8法、伤口愈合试验、细胞迁移和侵袭试验评估UNC1999与索拉非尼的协同作用。进行RNA测序（RNA-seq）以筛选UNC1999、索拉非尼及联合治疗后Huh7和SMMC7721细胞系中的差异表达基因。
在本研究中，miR-20a在HCC组织和细胞系中均呈低表达。miR-20a抑制SMMC7721和Huh7细胞的增殖和迁移。荧光素酶报告基因检测和蛋白质印迹分析结果显示，miR-20a直接靶向组蛋白甲基转移酶EZH1。我们证明miR-20a通过降低H3K27甲基化负调控EZH1的表达并抑制HCC的增殖和转移。我们发现UNC1999抑制肿瘤细胞增殖并增强索拉非尼的抑制作用。
我们证明miR-20a通过直接靶向EZH1抑制HCC的肿瘤增殖和转移。UNC1999可抑制肿瘤增殖并增加肝癌细胞系对索拉非尼的敏感性。
