Exploration of Mechanisms of Keliu Pill on the Inhibition of Lymphangiogenesis in Non-small Cell Lung Cancer Based on Network Pharmacology and Experimental Validation.

Keliu Pill (KLW) is a Chinese medicine formula that has been shown to be effective in treating non-small cell lung cancer (NSCLC) patients. However, the potential molecular mechanism remains unclear. To explore the underlying mechanism of Keliu Pill (KLW) in treating NSCLC, the network pharmacology was applied to explore the potential mechanism of KLW in the treatment of lung cancer. With oral bioavailability (OB) ≥ 30% and drug-like index (DL) ≥ 0.18 as the filter criteria, the compounds of 6 traditional Chinese medicines (TCMs) from KLW were retrieved on Traditional Chinese Medicine Systematic Pharmacology Database and Analysis Platform (TCMSP) and supplemented by Traditional Chinese Medicine Comprehensive Database (TCMID). The potential targets corresponding to the active components of the TCMs mentioned above were obtained from TCMSP. Subsequently, the potential underlying action mechanisms of KLW on NSCLC predicted by the network pharmacology analyses were experimentally validated in Lewis lung cancer cell inoculation mice models and co-culture models with Lewis lung cancer cell and tumor-associated macrophages (TAMs). After database search and screening, a total of 20 active ingredients of Astragalus Membranaceus, 21 of Sophare Tonkinensis Radix et Rhizoma, 1 of Mylabris, 3 of Eupolyphaga, 1 of Leech, and 3 of Gecko were obtained. After deleting duplicates, 49 active ingredients were obtained. Thirty important active ingredients in KLW were sorted out by referring to relevant literature. The KEGG pathway enrichment analysis based on the DAVID platform showed that the VEGF-C signaling pathway may be the core signaling pathway associated with KLW in the treatment of NSCLC. The network pharmacological analysis demonstrated that the various components of KLW acted on NSCLC through 29 potential targets such as VEGF-C, VEGF-D, MAPK8, MAPK1, and AKT1. The experiments in vivo indicated KLW could inhibit the transcription and translation of VEGF-C and VEGF-D in tumor tissues. KLW could reduce the proportion of activated TAMs in the tissue. KLW-contained serum may decrease the expression level of VEGF-C and VEGF-D and inhibit the lymphatic lumen formation of human lymphatic endothelial cells. VEGFC and VEGFD were confirmed as the potential KLW-associated targets for NSCLC. KLW may inhibit lymphatic angiogenesis of lung cancer by regulating the function of TAMs.