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Objective: From the perspective of the tumor microenvironment, this study explored the anti-tumor mechanisms of Ganoderma lucidum, Phellinus linteus, and Cordyceps militaris. Methods: Aqueous extracts (FAEs) of Ganoderma lucidum (RL), Phellinus linteus (BJ), and Cordyceps militaris (XG) were prepared via water bath extraction and vacuum freeze-drying. The effects of the FAEs and their conditioned media (CM) on the proliferative activity of A549 non-small cell lung cancer cells were determined using the MTT assay and Annexin V-FITC/PI double staining. Transcriptome sequencing was performed on cells treated with 4 mg/mL FAE, and KEGG pathway enrichment analysis was used to identify the response characteristics of the IL-17 signaling pathway to each extract. Additionally, immunocytochemical staining, antibody blocking assays, and ELISA were used to detect the effects of the FAEs on the expression levels of the mucins MUC5AC and MUC5B and the chemokines IL-6, CCL2, CCL20, CXCL8, and CXCL10. Results: The three FAEs and CM inhibited the proliferation of the A549 cells and induced their apoptosis. Transcriptome analysis results revealed that upregulated genes were predominant in the RL and BJ groups, whereas downregulated genes were predominant in the XG group. All groups showed significant enrichment in signaling pathways related to tumor microenvironment formation. Additionally, significant alterations were observed in the expression levels of key effector molecules downstream of the IL-17 signaling pathway (ko04657) in all groups. All three FAEs significantly inhibited MUC5B expression (P<0.01). However, BJ treatment did not significantly affect MUC5AC expression (P>0.05). MUC5AC antibody blocking weakened the inhibitory effects of RL and XG on cell proliferation (P<0.05), whereas MUC5B antibody blocking significantly reduced the anti-proliferative effects of RL and BJ on the cells (P<0.05). Furthermore, all three FAEs significantly inhibited the secretion of IL-6, CCL2, CCL20, and CXCL8 (P<0.01). Conclusion: In conclusion, Ganoderma lucidum, Phellinus linteus, and Cordyceps militaris can all remodel the tumor microenvironment to synergistically exert anti-tumor effects. This provides a new theoretical basis for their development as functional foods for adjuvant cancer therapy.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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