The incidence of gastric cancer is extremely high worldwide, and the incidence of gastric cancer in China is much higher than that in many other countries. TCM plays an important role in the prevention and treatment of tumors, mainly in the mechanisms of action that enhance the body's immune function, inhibit tumor cells division and proliferation, accelerate tumor cells apoptosis, and reverse multi-drug resistance of tumor cells [25].However, the composition of traditional Chinese medicines is complex, and it is difficult to elucidate their mechanisms of action due to the diversity of herbal components and action targets, thus lacking reliable scientific evidence to verify their effectiveness [26].In recent years, with the rapid development of high-throughput technology and bioinformatics, virtual pharmacology studies are considered to be the fastest and most effective screening method for early studies of drug effectiveness, solving the problem of multi-component, multi-target, and complex diseases in TCM [27]. In this study, the drug-component-intersection target-disease network diagram and the associated PPI network diagram were constructed, along with GO and KEGG enrichment analysis, to systematically elucidate the mechanism of action of Jin Ling Zi Powder in the treatment of gastric cancer. Molecular docking was used to verify the correspondence between targets and components, and good docking scores reflected the effectiveness of the components.
According to the network pharmacology research, there are 55 active ingredients in Jin Ling Zi Powder, and the main active ingredients including quercetin, (S)-Scoulerine, Isocorypalmine, leonticine, Hyndarin, etc.Haghi A et al. found that quercetin inhibits cell growth and induces apoptosis, necrosis and autophagy through and anti-Helicobacterial activity for the treatment of gastric cancer [28].Wangchuk P et al. found that (S)-Scoulerine achieved the treatment of cancer and inflammation through the inhibition of acetylcholinesterase, TNF-α, and Helicobacter pylori [29].Although the TCMSP database shows that Isocorypalmine, leonticine, and Hyndarin all have anti-cancer effects, their related mechanisms of action are unclear.This study shows that the above drug components may achieve their effects in the treatment of gastric cancer through IL6, PTGS2, MMP9, HMOX1, MYC, and other targets, which may provide a mechanism of action The present study showed that the above drug components may be used to treat gastric cancer through IL6, PTGS2, MMP9, HMOX1, MYC and other targets, which can provide reference for research on its action mechanism.
The target screening by PPI network topology analysis diagram, drug-component-intersection target-disease network diagram, and pathway-target network diagram suggested that Jin Ling Zi Powder for the treatment of gastric cancer might be closely related to such targets as IL6, PTGS2, MMP9, HMOX1, MYC, CHRM3, TOP2A, CA2, and KCNMA1.Huang SP et al. found that IL-6 expression was significantly elevated in gastric cancer tissues, which could promote angiogenesis and affect tumor cells adhesion and invasion by inducing the synthesis of vascular endothelial growth factor [30]. IL-6 was found to be one of the most important cytokine families in tumorigenesis and metastasis and was highly expressed in gastric cancer tissues [31].Meanwhile, a study found that the PD-L1 glycosylation process mediated by IL-6 triggered by STT3A not only upregulated the expression level of PD-L1 in tumor tissues, but also reduced the detection rate of PD-L1, resulting in some patients missing the opportunity of immune-checkpoint-blockade (ICB) therapy for a more prolonged survival benefit [32]. Sun WH et al. found that COX-2 (PTGS2) was not expressed in normal gastric mucosal tissues but was significantly more expressed in gastric cancer tissues [33], possibly by promoting angiogenesis and lymph node metastasis [34], immune escape [35], altering telomerase activity [36], acting on adhesion factors [37], inhibiting apoptosis [38 and romoting cell proliferation [39] are among the mechanisms that promote the development of gastric cancer.Torii A et al. found that MMP-9, a protease closely related to gastric cancer invasion and metastasis, caused gastric cancer invasion and metastasis by degrading the extracellular matrix [40].Ren QG et al. found that HMOX1 expression was elevated in gastric cancer tissues and played a crucial role in the development of gastric cancer. It is closely related to malignant apoptosis, immune escape and oxidative stress, etc.And low HMOX1 expression promotes gastric cancer cells apoptosis, inhibits their proliferation and invasion, improves overall patient survival, and may become an important target for the treatment of gastric cancer [41].The MYC gene consists of three paralogs, C-MYC, N-MYC and L-MYC, and is one of the most frequently dysregulated driver genes in human cancer that can disrupt the microenvironment of tumor cells and evade the immune response [42].Liu M et al. found that MYC is highly expressed in gastric cancer tissues, that its expression level is negatively correlated with clinical outcome, and that the development of gastric cancer requires a novel mechanism of transcriptional repression dependent on c-Myc target genes, providing a potential new strategy for targeted therapy of gastric cancer [43].CHRM3 is one of the M receptors, a G protein-coupled receptor involved in fluid-secreting exocrine gland cells, consisting of seven transmembrane structural domains that are expressed in a variety of tumor cells and can be linked to a variety of signaling pathways to play a role in promoting tumor cell proliferation and metastasis [44].Sorokin M et al. found that CHRM3 is also closely related in immunotherapy of gastric cancer [45].Wang Y et al. found that TOP2A is a key gene in gastric carcinogenesis [46] and can inhibit the proliferation, migration and invasion of gastric cancer cells and promote apoptosis by decreasing TOP2A expression [47].CAII has been shown to be associated with gastrointestinal tumors, with much less expression in malignant tissues and a negative decrease with increasing tumor malignancy, suggesting that they may be associated with tumor progression and invasion [48].Ma G found that KCNMA1 is a key oncogene in gastric carcinogenesis and its hypermethylation is an independent factor affecting the prognosis of gastric cancer patients [49]. In conclusion, in terms of action targets, Jin Ling Zi Powder for gastric cancer may be related to oxidative stress, cell proliferation and apoptosis, immune response, and inhibition of angiogenesis. Molecular docking also verified that the main active ingredients of Jin Ling Zi Powder for the treatment of gastric cancer all had good binding ability with their targets, which further improved the credibility of the results of this network pharmacology study.
The top ranking entries of GO functional enrichment analysis results suggest that Jin Ling Zi Powder for gastric cancer is mainly related to inflammatory response, cellular response to tumor necrosis factor, etc. Inflammatory response has been an important influencing factor in tumorigenesis and development, and the tumor microenvironment mediated by inflammatory cells is an indispensable player in tumor formation, promoting proliferation, survival and migration of tumor cells [50]. Tumor cells are also involved in the innate immune system through some signaling molecules such as selectins, chemokines and their receptors in order to invade, migrate and metastasize [51]. Not only that, Balkwill F found that TNF is an important inflammatory cytokine that causes hemorrhagic necrosis of tumor tissue by increasing vascular permeability and causing other types of cells in the blood to spill through the cell membrane, resulting in the loss of a large number of red blood cells in the blood vessels [52]. It is thus hypothesized that Jin Ling Zi Powder may achieve the treatment of gastric cancer by reducing the inflammatory response and inducing the cellular response to tumor necrosis factor, among other ways.
The KEGG pathway enrichment analysis revealed that the process of Jin Ling Zi Powder for gastric cancer mainly involved AGE-RAGE signaling pathway in diabetic complications, TNF signaling pathway, MicroRNAs in cancer, Pathways in cancer, etc. It was found that RAGE is a factor that regulates cancer cells invasion and metastasis, that RAGE expression is closely related to invasion and metastasis of gastric cancer [53], and that the AGE-RAGE system is a new target for the treatment of various tumors [54]. The TNF-α/IL-33/ST2L signaling-mediated epithelial-stromal interaction was found to play a key role in the progression of gastric cancer and provided the rationale for targeting this pathway for the treatment of gastric cancer metastasis [55]. Immunotherapy offers new hope for gastric cancer patients,macrophages infiltrating gastric cancer tissues regulate PD-L1 expression by releasing the pro-inflammatory cytokine TNF-ɑ, which regulates PD-L1 expression by activating NF-kB and STAT3 signaling pathways, and the expression of TNF-ɑ has important prognostic value in gastric cancer [56].MicroRNAs are stably present in serum and tissues and are key molecule in post-transcriptional regulation of gene expression, which can regulate gastric cancer cells survival and apoptosis, cell proliferation, invasion and metastasis, and is the hub of gene regulation in the process of gastric cancer development and progression [57], and microRNA can affect the sensitivity of gastric cancer patients to chemotherapy through multiple pathways, and microRNA-based therapy is considered a suitable approach to overcome chemotherapy resistance in gastric cancer patients [ 58].Pathways in cancer play different roles in different types of tumors, and in gastric cancer, the synergistic dysregulation of multiple Pathways in cancer may activate signature overgrowth, which in turn leads to the development of gastric cancer [59]. In conclusion, Jin Ling Zi Powder may play a role in the treatment of gastric cancer through the frontal regulation of immune, drug resistance, multi-target and multi-pathway pathways.
The molecular docking results showed that the binding energy between all three compounds and the target protein was less than 5 kJ/mol, indicating that these core compounds have high binding activity to the receptor protein, further validating the results of the network pharmacology study.
However, there are limitations in the network pharmacology analysis, which in most studies used two parameters of OB and DL compounds from the herbal database, while low DL compounds were often ignored [60]. In subsequent studies, these data need to be further experimentally validated at the cellular, animal, and molecular levels to provide an experimental basis for the treatment of gastric cancer with Jin Ling Zi Powder.