Tan Wenliang, Lin Zhirong, Chen Xianqing, Li Wenxin, Zhu Sicong, Wei Yingcheng, Huo Liyun, Chen Yajin, Shang Changzhen
Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.
Department of Hepatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.
Ann Transl Med. 2021 Jan;9(1):38. doi: 10.21037/atm-20-2081.
Sorafenib can prolong the survival of patients with advanced hepatocellular carcinoma (HCC). However, drug resistance remains the main obstacle to improving its efficiency. This study aimed to explore the likely molecular mechanism of sorafenib resistance.
Differentially expressed microRNAs (miRNAs) related to sorafenib response were analyzed with the Limma package in R software. The expression levels of miR-126-3p and sprouty-related EVH1 domain-containing protein 1 (SPRED1) in HCC cells were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell viability and proliferation were detected with Cell Counting Kit-8 (CCK-8), EdU proliferation, and clone formation assays. Transwell assays were performed to measure cell migration and invasion. TargetScan, MicroRNA Target Prediction Database (miRDB), and StarBase v2.0 were used to predict the targets of miR-126-3p. SPRED1 was confirmed as a target gene of miR-126-3p by dual-luciferase reporter assay and Western blotting. Finally, the anti-tumor effect of LV-miR-126-3p inhibitor combined with sorafenib was evaluated via subcutaneous tumor models.
HCC cells with high expression of miR-126-3p exhibited increased resistance to sorafenib. The results of bioinformatics analysis and the dual-luciferase reporter assay showed that miR-126-3p directly targeted SPRED1. The sensitivity of HCC cells to sorafenib was markedly enhanced by SPRED1 upregulation. Gain- and loss-of function experiments verified that miR-126-3p induced sorafenib resistance in HCC through downregulating SPRED1. Furthermore, the inhibition of miR-126-3p markedly increased the effectiveness of sorafenib against HCC . Mechanistically, our results suggested that miR-126-3p promoted sorafenib resistance via targeting SPRED1 and activating the ERK signaling pathway.
Our study demonstrates that regulating the miR-126-3p/SPRED1 axis might be a promising strategy for enhancing the antitumor effect of sorafenib in the treatment of HCC.
索拉非尼可延长晚期肝细胞癌（HCC）患者的生存期。然而，耐药性仍然是提高其疗效的主要障碍。本研究旨在探讨索拉非尼耐药可能的分子机制。
使用R软件中的Limma软件包分析与索拉非尼反应相关的差异表达微小RNA（miRNA）。通过定量逆转录聚合酶链反应（qRT-PCR）检测HCC细胞中miR-126-3p和含Sprouty相关EVH1结构域蛋白1（SPRED1）的表达水平。用细胞计数试剂盒-8（CCK-8）、EdU增殖和克隆形成试验检测细胞活力和增殖。进行Transwell试验以测量细胞迁移和侵袭。使用TargetScan、微小RNA靶标预测数据库（miRDB）和StarBase v2.0预测miR-126-3p的靶标。通过双荧光素酶报告基因试验和蛋白质印迹法确认SPRED1是miR-126-3p的靶基因。最后，通过皮下肿瘤模型评估LV-miR-126-3p抑制剂联合索拉非尼的抗肿瘤作用。
miR-126-3p高表达的HCC细胞对索拉非尼的耐药性增加。生物信息学分析和双荧光素酶报告基因试验结果表明，miR-126-3p直接靶向SPRED1。SPRED1上调显著增强了HCC细胞对索拉非尼的敏感性。功能获得和缺失实验证实，miR-126-3p通过下调SPRED1诱导HCC细胞对索拉非尼产生耐药性。此外，抑制miR-126-3p显著提高了索拉非尼对HCC的疗效。机制上，我们的结果表明，miR-126-3p通过靶向SPRED1并激活ERK信号通路促进索拉非尼耐药。
我们的研究表明，调节miR-126-3p/SPRED1轴可能是增强索拉非尼治疗HCC抗肿瘤作用的一种有前景的策略。
