Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang, China.
Key Laboratory of Structure-Based Drug Design and Discovery, Ministry of Education, Shenyang Pharmaceutical University, Shenyang, China.
J Pharm Pharmacol. 2020 Oct;72(10):1370-1382. doi: 10.1111/jphp.13315. Epub 2020 Jun 28.
To examine the antiproliferative effects of 1-(4-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)-3-(2-(dimethylamino)ethyl)imidazolidin-2-one (ZX-42) on the echinoderm microtubule-associated protein-4/anaplastic lymphoma kinase fusion gene (EML4-ALK) positive lung cancer cell line H2228 and its underlying mechanism.
The MTT assay was used to study the effect of ZX-42 on H2228 cell growth. Propidium iodide (PI) staining and Western blotting were used to investigate the cell cycle changes. ZX-42-induced cell apoptosis was determined using the Annexin V-FITC/PI (AV/PI) apoptotic assay kit, acridine orange/ethidium bromide (AO/EB) and Hoechst 33258 staining, Rhodamine 123 (Rh 123) fluorescence assay and Western blotting. ZX-42-induced reactive oxygen species (ROS) production was examined by ROS assay kit. Transmission electron microscope, monodansylcadaverine (MDC) staining and the AV/PI apoptotic assay kit were used to demonstrate the relationship between autophagy and apoptosis.
ZX-42 had good cell viability inhibitory effect on H2228 cells. ZX-42 dramatically inhibited ALK and its downstream pathways. ZX-42 also blocked H2228 cell cycle at G1 phase and then induced apoptosis by activating the mitochondrial pathway. Next, ZX-42 induced the production of ROS, and antioxidant N-acetylcysteine (NAC) reduced ROS production and also decreased apoptotic rates. We also found that ZX-42 induced protective autophagy in H2228 cells.
In summary, ZX-42 is a novel ALK inhibitor that significantly inhibits the cell viability of H2228 cells and ultimately induces apoptosis through the mitochondrial pathway, in which autophagy plays a protective role. Therefore, inhibition of autophagy might enhance the anti-cancer effect of ZX-42.
研究 1-(4-((5-氯-4-((2-(异丙基磺酰基)苯基)氨基)嘧啶-2-基)氨基)-3-甲氧基苯基)-3-(2-(二甲基氨基)乙基)咪唑烷-2-酮（ZX-42）对棘皮动物微管相关蛋白-4/间变性淋巴瘤激酶融合基因（EML4-ALK）阳性肺癌细胞系 H2228 的抗增殖作用及其机制。
采用 MTT 法研究 ZX-42 对 H2228 细胞生长的影响。碘化丙啶（PI）染色和 Western blot 法检测细胞周期变化。采用 Annexin V-FITC/PI（AV/PI）凋亡试剂盒、吖啶橙/溴化乙锭（AO/EB）和 Hoechst 33258 染色、罗丹明 123（Rh 123）荧光法和 Western blot 法检测 ZX-42 诱导的细胞凋亡。采用 ROS 试剂盒检测 ZX-42 诱导的活性氧（ROS）产生。透射电镜、单丹磺酰尸胺（MDC）染色和 AV/PI 凋亡试剂盒用于证明自噬与凋亡之间的关系。
ZX-42 对 H2228 细胞具有良好的细胞活力抑制作用。ZX-42 显著抑制 ALK 及其下游通路。ZX-42 还通过阻断 H2228 细胞周期 G1 期，然后通过激活线粒体途径诱导细胞凋亡。接下来，ZX-42 诱导 ROS 产生，抗氧化剂 N-乙酰半胱氨酸（NAC）减少 ROS 产生，也降低凋亡率。我们还发现，ZX-42 诱导 H2228 细胞保护性自噬。
总之，ZX-42 是一种新型的 ALK 抑制剂，可显著抑制 H2228 细胞的细胞活力，并通过线粒体途径最终诱导细胞凋亡，其中自噬起保护作用。因此，抑制自噬可能增强 ZX-42 的抗癌作用。
