Department of Minimal Invasive Surgery, the Second Xiangya Hospital of Central South University, Changsha, 410000, China.
Department of Institute of Hepatobiliary Diseases, the Zhongnan Hospital of Wuhan University, Wuhan, 430000, China.
Cell Biol Int. 2020 Feb;44(2):467-476. doi: 10.1002/cbin.11247. Epub 2019 Nov 4.
Shikonin is a natural naphthoquinone component with antioxidant and anti-tumor function and has been used for hepatocellular carcinoma (HCC) treatment. According to the previous study, many herbs can regulate cancer cell progression by targeting specific microRNA (miRNA) (Liu, 2016). However, the underlying pathological mechanism of shikonin in HCC therapy is still unclear. The detection of cell growth and death rate were performed by hemacytometry and trypan blue staining, respectively. The expression of miR-106b and SMAD7 messenger RNA (mRNA) in HCC cells was evaluated by quantitative real-time polymerase chain reaction. Cell proliferation, apoptosis, and migration ability were measured by cell counting kit-8 (CCK-8), flow cytometry, and transwell assay. The expression of proteins E-cadherin, N-cadherin, vimentin, SMAD7, TGF-β1, p-SMAD3, SMAD3, and GAPDH was examined by western blot. The interaction between SMAD7 and miR-106b was assessed by luciferase reporter system. Shikonin inhibited Huh7 and HepG2 cell growth in a dose-dependent manner while induced cell death in a time-dependent manner. In addition, the expression of miR-106b was reduced after shikonin treatment. Moreover, miR-106b attenuated the suppressive effects of shikonin on HCC cell migration and epithelial-mesenchymal transition (EMT). SMAD7 was predicted as a target of miR-106b and the prediction was confirmed by luciferase reporter system. Additionally, we observed that SMAD7 reversed the promotive effects of miR-106b on HCC cell progression and EMT. The subsequent western blot assay revealed that shikonin could modulate SMAD7/TGF-β signaling pathway by targeting miR-106b. In conclusion, Shikonin suppresses cell progression and EMT and accelerates cell death of HCC cells via modulating miR-106b/SMAD7/TGF-β signaling pathway, suggesting shikonin could be an effective agent for HCC treatment.
紫草素是一种具有抗氧化和抗肿瘤功能的天然萘醌类成分，已用于肝细胞癌（HCC）的治疗。根据之前的研究，许多草药可以通过靶向特定的 microRNA（miRNA）来调节癌细胞的进展（Liu，2016）。然而，紫草素在 HCC 治疗中的潜在病理机制尚不清楚。通过血细胞计数器和台盼蓝染色分别检测细胞生长和死亡率。通过实时定量聚合酶链反应评估 HCC 细胞中 miR-106b 和 SMAD7 信使 RNA（mRNA）的表达。通过细胞计数试剂盒-8（CCK-8）、流式细胞术和 Transwell 测定法测量细胞增殖、凋亡和迁移能力。通过 Western blot 检测 E-钙粘蛋白、N-钙粘蛋白、波形蛋白、SMAD7、TGF-β1、p-SMAD3、SMAD3 和 GAPDH 蛋白的表达。通过荧光素酶报告系统评估 SMAD7 和 miR-106b 之间的相互作用。紫草素呈剂量依赖性抑制 Huh7 和 HepG2 细胞生长，同时呈时间依赖性诱导细胞死亡。此外，紫草素处理后 miR-106b 的表达减少。此外，miR-106b 减弱了紫草素对 HCC 细胞迁移和上皮-间充质转化（EMT）的抑制作用。SMAD7 被预测为 miR-106b 的靶标，并且通过荧光素酶报告系统证实了该预测。此外，我们观察到 SMAD7 逆转了 miR-106b 对 HCC 细胞进展和 EMT 的促进作用。随后的 Western blot 检测表明，紫草素可以通过靶向 miR-106b 来调节 SMAD7/TGF-β 信号通路。总之，紫草素通过调节 miR-106b/SMAD7/TGF-β 信号通路抑制 HCC 细胞的进展和 EMT，加速 HCC 细胞的死亡，提示紫草素可能是 HCC 治疗的有效药物。
