School of Medicine, Zhejiang University City College, No.51 Huzhou Street, Hangzhou, Zhejiang, 310015, China,; College of Pharmaceutical Sciences, Zhejiang University, No.866 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China.
School of Medicine, Zhejiang University City College, No.51 Huzhou Street, Hangzhou, Zhejiang, 310015, China.
Phytomedicine. 2020 Mar;68:153189. doi: 10.1016/j.phymed.2020.153189. Epub 2020 Feb 10.
NSCLC is the major type of lung cancer and the survival rates of NSCLC patients remain low. AZD9291 is a third-generation EGFR-TKI and approved to treat NSCLC patients harboring EGFR T790M mutation and common targetable activating EGFR mutations, but it has a limited effect for wtEGFR NSCLC.
The current study investigated whether shikonin could enhance the antitumor effect of AZD9291 in wtEGFR NSCLC cells.
SRB and colony formation assay were used to detect the proliferation of NSCLC cells, propidium iodide staining was performed to detect the apoptosis, ROS was analyzed using DCFH-DA staining, and western blot was used to detect the expression of indicated proteins.
We demonstrated that shikonin, a natural ROS inducer, could enhance the antitumor effect of AZD9291 in wtEGFR NSCLC cells. In addition, shikonin increased AZD9291-induced apoptosis accompanying with the generation of ROS and activation of ER stress. Furthermore, ROS inhibition by NAC or GSH reversed the apoptosis induced by shikonin plus AZD9291, and recovered the ER stress activated by combination treatment, indicating that ROS mediated ER stress played a vital role in this combination therapy. Moreover, shikonin increased the anticancer activity of AZD9291 in primary wtEGFR NSCLC cells through ROS-mediated ER stress.
Our study suggests that combining shikonin with AZD9291 is a promising therapeutic strategy for treating wtEGFR NSCLC patients.
非小细胞肺癌（NSCLC）是肺癌的主要类型，其患者的生存率仍然较低。AZD9291 是一种第三代 EGFR-TKI，被批准用于治疗携带 EGFR T790M 突变和常见可靶向激活 EGFR 突变的 NSCLC 患者，但对 wtEGFR NSCLC 的疗效有限。
本研究旨在探讨紫草素能否增强 AZD9291 对 wtEGFR NSCLC 细胞的抗肿瘤作用。
采用 SRB 和集落形成实验检测 NSCLC 细胞的增殖，碘化丙啶染色检测细胞凋亡，DCFH-DA 染色分析 ROS 水平，Western blot 检测相关蛋白的表达。
我们发现，紫草素作为一种天然的 ROS 诱导剂，能够增强 AZD9291 对 wtEGFR NSCLC 细胞的抗肿瘤作用。此外，紫草素增加了 AZD9291 诱导的凋亡，伴随着 ROS 的产生和 ER 应激的激活。进一步的研究表明，ROS 抑制剂 NAC 或 GSH 逆转了紫草素加 AZD9291 诱导的细胞凋亡，并恢复了联合治疗激活的 ER 应激，表明 ROS 介导的 ER 应激在这种联合治疗中发挥了重要作用。此外，紫草素通过 ROS 介导的 ER 应激增强了 AZD9291 在原发性 wtEGFR NSCLC 细胞中的抗癌活性。
我们的研究表明，将紫草素与 AZD9291 联合使用可能是治疗 wtEGFR NSCLC 患者的一种有前途的治疗策略。
