According to the latest worldwide cancer statistics, lung cancer has been reported a significant increase in morbidity and mortality, which accounts for the leading cause of threatening the health and life of human being(Siegel et al., 2018). Because of the aging population and the huge population, China has an extremely high incidence of lung tumors.(Bray et al., 2018). Of all the lung cancer histological subtypes, non-small cell lung cancer (NSCLC) is the most common. A significant proportion of patients receive their diagnosis during the advanced stages of the disease, which is associated with a prognostic indicator of a low 5-year overall survival rate(Atsaves et al., 2019). Traditional treatments for lung cancer, which include surgery, chemotherapy, radiation, and targeted therapy, have significantly improved the prognosis (Moya-Horno et al., 2018). Immune checkpoint inhibitors have emerged in recent years to reverse NSCLC immunosuppression and enhance antitumor immunity, including programmed cell death protein-1(PD-1) and its ligand (PD-L1). Despite various anti-PD-1/PD-L1 antibodies are currently used as first and second-line treatment for NSCLC patients, less than 30% of the lung cancer population benefit from this therapy(Li et al., 2018). Unmanageable toxicity, including immune hepatitis, immune thyroiditis, immune pneumonia and reactive capillary hyperplasia are commonly encountered, has impacted on the therapeutic effect seriously(Kennedy and Salama, 2020). Therefore, it is imperative to search for therapeutic drugs that exhibit low side effects and high efficacy in the NSCLC.

With over 3500 years of experience, traditional Chinese medicine (TCM) has been utilized to treat a broad spectrum of illnesses, including cancer. As a complementary and alternative therapy, TCM offers a promising treatment for lung cancer. Numerous clinical studies have confirmed that the single herb and formula form may both successfully boost immune system performance, reduce clinical symptoms, and increase survival time while having fewer side effects(Li et al., 2021). TCM anti-tumor has become a popular research direction in recent years, and its anti-tumor mechanisms have been revealed by more scientists. Polygonatum cyrtonema Hua (Huangjing) is a perennial herb of the family asparagaceae. Its name has been checked with “World Flora Online” (www.worldfloraonline.org). And we can learn more about Huangjing by visiting this website (https://www.worldfloraonline.org/taxon/wfo-0000716205). Huangjing is one of the representative medicinal plants of the Lung Meridian and is widely used in the clinical treatment of lung diseases because of its traditional effects of tonifying the lungs and nourishing the yin. Also Huangjing is recognized as a representative example of the concept of "medicine food homology" (MFH). It has been traditionally employed for its therapeutic properties in the management of cancer and as a means to enhance lung function. Recently, it is in evidence that Huangjing possesses anti-neoplastic and anti-inflammatory effects. For instance, Huangjing extract has been featured that it can inhibit tumor growth and improve the spleen index, thymus index. Meanwhile, there was a statistically significant increase in the cytokines secretion and CD4+/CD8+ lymphocytes ratio(Long et al., 2018). It has been also found that Polygonatum odoratum lectin inhibiting Wnt pathway to exert an anti-neoplastic effect(Wu et al., 2016). Anti-lung cancer effects of Huangjing extract in inhibiting the TLR4-MAPK/NF-κB signaling pathways [8]. Although, all of the above studies suggest that Huangjing may as a remarkable anti-lung tumor role, the underlying molecular action mechanism of Huangjing is also still not yet known.

Network pharmacology is a developing field of study that investigates the interconnectedness of diseases, therapeutic targets, and medications by analyzing network data. It aims to get an understanding of the molecular mechanisms behind the anticancer effects of active components found in TCM. Through the retrieval of databases pertaining to Chinese herbs, compounds, and genes, the objective of attaining a first comprehension of each formula is accomplished. Cytoscape was used to analyze protein-to-protein interaction and route enrichment to construct a comprehensive network. One notable advantage of network pharmacological analysis is in its ability to facilitate direct verification tests based on the explicit network outcomes(Xie et al., 2019).

In this study, we investigated the mechanisms of suppression of PI3K/Akt/HIF-1α signaling by Huangjing in NSCLC by means of network pharmacology, molecular docking, and experimental verification. Firstly, network pharmacology was utilized to predict the potential interaction between components from Huangjing, targets and relevant pathways involved in NSCLC. To clarify whether Huangjing has antitumor properties, Lewis lung carcinoma (LLC) tumor-bearing mice model was conducted. We also carried out tissue quantitative analysis of hub genes. Next, lung adenocarcinoma cellline A549 cells were treated with active ingredients β-sitosterol, then apoptosis was evaluated by CCK-8, colony formation assay, and flow cytometry. Western blot and quantitative real-time PCR were performed to detect the expression of target genes. Our findings suggest Huangjing may be a supplementary treatment for NSCLC patients.