Sineoculis Homeobox Homolog 1（SIX1）基因通过影响人肝癌细胞系 HepG2 中的活性氧和自噬来调节紫杉醇耐药性。

The Sineoculis Homeobox Homolog 1 (SIX1) Gene Regulates Paclitaxel Resistance by Affecting Reactive Oxygen Species and Autophagy in Human Hepatocellular Carcinoma Cell Line HepG2.

机构信息

Department of Radiology, Affiliated Hospital of Hebei University of Engineering, Handan, Hebei, China (mainland).

Department of Radiology, The First Hospital of Yongnian District, Handan, Hebei, China (mainland).

出版信息

Med Sci Monit. 2018 Apr 15;24:2271-2279. doi: 10.12659/msm.906361.

BACKGROUND The objective of this study was to explore the role of SIX1 in paclitaxel (TAX) resistance of HepG2 cells via reactive oxygen species (ROS) and autophagy pathway. MATERIAL AND METHODS Hepatoma cell line HepG2 was treated with SIX1 knockdown or/and TAX. Cell growth was detected by MTT assay and colony formation assay. Cell apoptosis was evaluated with flow cytometry. ROS levels were detected using flow cytometry (stained with DCFH2-DA). Western blot was conducted to detect the expression of SIX1 and autophagy-related proteins. RESULTS TAX suppressed the proliferation of HepG2 cells in a time/dose-dependent manner, and upregulated the expression of SIX1. SIX1 siRNA increased TAX sensitivity of HepG2 cells and upregulated cell ROS levels. SIX1 siRNA combined with TAX treatment activated autophagy of HepG2 cells. N-acetyl-L-cysteine (NAC) partially attenuated SIX1 siRNA-induced ROS level increases, and autophagy inhibitor 3-MA notably enhanced SIX1 siRNA-induced cell apoptosis. CONCLUSIONS Knockdown of SIX1 increased cell ROS levels and autophagy, promoted cell apoptosis, and enhanced TAX sensitivity of HepG2 cells.

背景

本研究旨在探讨 SIX1 通过活性氧（ROS）和自噬途径在紫杉醇（TAX）耐药的 HepG2 细胞中的作用。

材料与方法

用 SIX1 敲低或/和 TAX 处理肝癌细胞系 HepG2。用 MTT 法和集落形成实验检测细胞生长。用流式细胞术评估细胞凋亡。用 DCFH2-DA 染色流式细胞术检测 ROS 水平。用 Western blot 检测 SIX1 和自噬相关蛋白的表达。

结果

TAX 呈时间和剂量依赖性抑制 HepG2 细胞增殖，并上调 SIX1 的表达。SIX1 siRNA 增加了 HepG2 细胞对 TAX 的敏感性，并上调了细胞 ROS 水平。SIX1 siRNA 联合 TAX 处理激活了 HepG2 细胞的自噬。N-乙酰-L-半胱氨酸（NAC）部分减弱了 SIX1 siRNA 诱导的 ROS 水平升高，而自噬抑制剂 3-MA 显著增强了 SIX1 siRNA 诱导的细胞凋亡。