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Objective: To identify the miRNAs that post-transcriptionally target the regulation of egfr in the antitumor mechanism of Cordyceps militaris. Methods: An aqueous extract of Cordyceps militaris was prepared using water-bath extraction combined with vacuum freeze-drying. Transcriptome sequencing was performed on A549 cells after 24 h treatment with 4 mg/mL Cordyceps militaris extract. Candidate miRNAs were selected on the basis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Functional gain models were constructed by transient transfection of miRNA mimics, and the effects of miRNAs on cell viability were assessed using the MTT assay and Annexin V-FITC/PI double staining. Functional loss models were established by transient transfection of miRNA inhibitors, in combination with Cordyceps militaris extract treatment. The effect of Cordyceps-miRNA axis on cell viability was evaluated using the MTT assay, and its regulatory effects on EGFR transcription and protein levels in A549 cells were analyzed using qRT-PCR and immunocytochemical staining. Results: A total of 7211 DEGs were obtained and significantly enriched in the "MicroRNAs in cancer" signaling pathway (Sort 3). Eleven miRNAs were screened for their association with lung adenocarcinoma prognosis, miR-125a-3p, miR-125a-5p, miR-125b-1-3p, miR-125b-2-3p, miR-125b-5p and miR-145-5p were all associated with egfr in a target-regulatory manner, and patients with lung adenocarcinomas that showed low expression levels had a poorer prognosis (P<0.05). Cordyceps militaris treatment induced significant upregulation of miR-125b-1-3p, miR-125b-5p and miR-145-5p expression levels (P<0.01), and the seed sequences had potential binding sites with the egfr 3'-UTR region. miR-125b-1-3p, miR-125b-5p, and miR-145-5p exhibited tumor-specific pro-proliferative activities, and their high expression status induced a higher percentage of early apoptosis in A549 cells. Cordyceps militaris inhibited the proliferation of lung cancer A549 cells by promoting the expression of miR-125b-1-3p, miR-125b-5p, and miR-145-5p (P<0.05) and downregulating the expression of EGFR (P<0.05). Conclusion: Cordyceps militaris exerts anti-tumor activity by targeting and regulating the miRNAs (miR-125b-1-3p, miR-125b-5p and miR-145-5p)-EGFR axis, providing theoretical support for its development as a novel functional food for adjuvant tumor 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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