miR-1226-3p Promotes Sorafenib Sensitivity of Hepatocellular Carcinoma via Downregulation of DUSP4 Expression
miR-1226-3p Promotes Sorafenib Sensitivity of Hepatocellular Carcinoma via Downregulation of DUSP4 Expression
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miR-1226-3p通过下调DUSP4表达促进肝细胞癌索拉非尼敏感性
DOI：
10.7150/jca.31804
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发表时间：
2019-01-01
影响因子：
3.9
通讯作者：
Chen, Yajin
中科院分区：
文献类型：
--
作者：
Chen, Xianqing;Tan, Wenliang;Chen, Yajin
背景：索拉非尼似乎可以提高肝细胞癌（HCC）患者的生存率，但由于耐药性，其缓解率受到严重限制。索拉非尼耐药的分子机制仍不清楚。在此，我们探讨了 miR-1226-3p 在 HCC 索拉非尼耐药中的可能作用。方法：通过 qRT-PCR 评估 HCC 细胞系中 miR-1226-3p 的表达水平。通过 CCK-8 测定测量细胞对索拉非尼的活力。通过流式细胞术和EdU增殖实验检测细胞凋亡和增殖。使用 DUSP4 3'-UTR 的荧光素酶报告基因作为 miR-1226-3p 的靶基因进行验证。最后通过裸鼠肝癌异种移植评价miR-1226-3p联合索拉非尼的体内抗肿瘤效果。结果：基因表达综合 (GEO) 数据集 GSE56059 的生物信息学分析表明，索拉非尼治疗后表现出疾病进展 (PD) 的 HCC 患者中 miR-1226-3p 表达下调。 SK-HEP-1 细胞表达的 miR-1226-3p 水平低于 HepG2 细胞。我们证实，与 HepG2 细胞相比，SK-HEP-1 细胞对索拉非尼更具耐药性。此外，miR-1226-3p模拟物增加SK-HEP-1细胞的细胞凋亡，而miR-1226-3p抑制剂在索拉非尼治疗后显着损害HepG2细胞的细胞凋亡。此外，我们验证了 miR-1226-3p 直接靶向双特异性磷酸酶 4 (DUSP4)，并进一步证明 DUSP4 的敲低通过调节 JNK-Bcl-2 轴来减少索拉非尼耐药。结论：miR-1226-3p通过下调DUSP4表达促进HCC对索拉非尼的敏感性，靶向miR-1226-3p可能是克服索拉非尼耐药的新治疗策略。
Background: Sorafenib appears to increase the survival rate of hepatocellular carcinoma (HCC) patients, but its response rate is seriously limited due to drug resistance. Molecular mechanisms underlying sorafenib resistance are still unknown. Herein, we explored the possible role of miR-1226-3p in sorafenib resistance of HCC. Methods: The miR-1226-3p expression level in HCC cell lines was evaluated by qRT-PCR. Cell viabilities to sorafenib were measured by CCK-8 assay. Cell apoptosis and proliferation were detected by flow cytometry and EdU proliferation assay. A luciferase reporter of DUSP4 3'-UTR was used for validation as a target gene of miR-1226-3p. Finally, the effects of in vivo antitumor efficacy of miR-1226-3p combined with sorafenib were evaluated by HCC tumor xenografts in nude mice. Results: Bioinformatics analysis from Gene Expression Omnibus (GEO) datasets GSE56059 suggested that miR-1226-3p expression was downregulated in HCC patients who showed progressive disease (PD) after sorafenib treatment. SK-HEP-1 cells expressed lower levels of miR-1226-3p than HepG2 cells. We confirmed that SK-HEP-1 cells were more resistant to sorafenib compared to HepG2 cells. In addition, miR-1226-3p mimic increased cell apoptosis of SK-HEP-1 cells, whereas miR-1226-3p inhibitor significantly impaired cell apoptosis of HepG2 cells after sorafenib treatment. Moreover, we validated that miR-1226-3p directly targeted dual specificity phosphatase 4 (DUSP4), and further demonstrated that knockdown of DUSP4 reduced sorafenib resistance by regulating the JNK-Bcl-2 axis. Conclusions: miR-1226-3p promotes sorafenib sensitivity of HCC through downregulation of DUSP4 expression, and targeting miR-1226-3p may be a novel therapeutic strategy for overcoming sorafenib resistance.