To investigate the anti-tumor activities of WZ35 and its possible molecular mechanism, bioinformatics analysis and the hematoxylin-eosin (HE) staining were applied to evaluate the Yes-associated-protein (YAP) level in gastric cancer. Cell counting kit-8 (CCK-8) was used to examine cell viability. Apoptosis was determined by flow cytometry analysis. Seahorse bioenergetics analyzer was used to investigate the alteration of oxygen consumption and aerobic glycolysis rate. SiRNA transfection was applied to silence endogenous YAP. Western blot was performed to detect indicated proteins. We found that treatment of gastric cancer cells with WZ35 exerted stronger anti-tumor activities than curcumin. Mechanistically, our research showed that WZ35 inhibited glycolysis, and induced reactive oxygen species (ROS) generation, resulting in Jun N-terminal Kinase (JNK) activation through downregulation of YAP in gastric cancer cells. ROS mediated YAP downregulation and JNK activation was regulated by glycolysis. Abrogation of ROS production markedly attenuated WZ35 induced anti-tumor activities as well as YAP downregulation and JNK activation. Similarly, the JNK inhibitor significantly reversed WZ35 induced anti-tumor activities in gastric cancer cells. Our study reveals a novel anti-gastric cancer mechanism of WZ35 by inhibiting glycolysis through the ROS-YAP-JNK pathway. WZ35 might be a potential therapeutics for the treatment of gastric cancer.

中文翻译：

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姜黄素类似物WZ35通过ROS-YAP-JNK途径影响胃癌细胞的糖酵解抑制作用。

为了研究WZ35的抗肿瘤活性及其可能的分子机制，应用生物信息学分析和苏木精-伊红（HE）染色评估了胃癌中Yes相关蛋白（YAP）的水平。细胞计数试剂盒8（CCK-8）用于检查细胞活力。通过流式细胞术分析确定细胞凋亡。使用海马生物能分析仪研究耗氧量和有氧糖酵解速率的变化。SiRNA转染应用于沉默内源性YAP。进行蛋白质印迹以检测指示的蛋白质。我们发现，用WZ35处理胃癌细胞比姜黄素具有更强的抗肿瘤活性。从机理上讲，我们的研究表明WZ35抑制了糖酵解，并诱导了活性氧（ROS）的产生，通过下调胃癌细胞中的YAP导致Jun N末端激酶（JNK）激活。ROS介导的YAP下调和JNK活化通过糖酵解来调节。活性氧生产的废止显着减弱了WZ35诱导的抗肿瘤活性以及YAP下调和JNK激活。同样，JNK抑制剂可显着逆转WZ35诱导的胃癌细胞的抗肿瘤活性。我们的研究通过抑制经由ROS-YAP-JNK途径的糖酵解揭示了WZ35的新型抗胃癌机制。WZ35可能是治疗胃癌的潜在疗法。活性氧生产的废止显着减弱了WZ35诱导的抗肿瘤活性以及YAP下调和JNK激活。同样，JNK抑制剂可显着逆转WZ35诱导的胃癌细胞的抗肿瘤活性。我们的研究通过抑制经由ROS-YAP-JNK途径的糖酵解揭示了WZ35的新型抗胃癌机制。WZ35可能是治疗胃癌的潜在疗法。活性氧生产的废止显着减弱了WZ35诱导的抗肿瘤活性以及YAP下调和JNK激活。同样，JNK抑制剂可显着逆转WZ35诱导的胃癌细胞的抗肿瘤活性。我们的研究通过抑制经由ROS-YAP-JNK途径的糖酵解揭示了WZ35的新型抗胃癌机制。WZ35可能是治疗胃癌的潜在疗法。