According to the World Health Organization's 2020 publication of global lung cancer incidence and mortality rates, lung cancer was the world's leading cause of cancer-related deaths and the second-most common cancer [1], [2]. The two primary subtypes of lung cancer were non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC), with NSCLC accounting for 80–85% of cases [3], [4]. Despite significant advances in treatment over the last decade, NSCLC has remained an incurable disease for most patients. Chemoradiation was the most effective treatment for patients with locally advanced but resectable NSCLC [5], [6]. However, most advanced NSCLCs resisted current therapies and eventually progressed [7]. Moreover, further efficacy was challenging due to the severe side effects and insufficient tolerability of chemoradiotherapy [8]. In recent years, gene-targeted therapy has expanded treatment options for patients with advanced NSCLC [9], [10]. Therefore, discovering new targeted agents for treating NSCLC was a pressing need.

Autophagy represents an intracellular degradation process that is evolutionarily conserved. This process involves ferrying damaged organelles, aggregated proteins, cytoplasmic macromolecules, or pathogens to lysosomes, where lysosomal hydrolases degrade them [11]. Recent studies have recognized autophagy as a form of programmed cell death type II, thereby revealing its significance in cell death mechanisms. Furthermore, autophagy has emerged as a potential therapeutic target for cancer treatment due to its influence on regulating cell death [12], [13]. Notably, the role of autophagy in cancer development is multifaceted and depends on the cancer type, developmental stage, and genetic background [14], [15]. During the initial stages of tumor development, autophagy serves as a critical factor in suppressing tumors by upholding genomic stability, averting cellular damage and inflammation, and restraining the accumulation of oncogenic p62 protein aggregates. Ultimately, this prevents tumorigenesis, proliferation, invasion, and metastasis [16], [17]. Studies investigating autophagy-linked signaling pathways have indicated that enhancing autophagy notably hinders the progression of NSCLC tumor cells. Both in clinical and preclinical stages, various medications targeting the mTOR, PI3K/AKT, and EGFR signaling pathways have demonstrated the capacity to activate autophagy and interfere with lung cancer cells [18], [19], [20]. Consequently, targeting autophagy-linked pathways and inciting autophagy could offer a promising treatment strategy for non-small cell lung cancer.

Previous studies found that 13-oxyingenol-dodecanoate (13OD) from Euphorbia kansui had good cytotoxic activity in A549 and H460 cells [21], [22]. However, the detailed mechanism and targets remained unknown. In this study, we investigated the antitumor activity of 13OD on NSCLC cells in vitro and in vivo. Subsequently, RNA-seq combined with Omics and Text-based Target Enrichment and Ranking (OTTER) methods were used to identify the target proteins of 13OD. The target proteins were validated using various biological methods, and the mechanism of action of 13OD in NSCLC cells was also investigated.