Network pharmacology and biological verification of morusin's therapeutic mechanisms in inhibiting nasopharyngeal carcinoma growth

Abstract

Nasopharyngeal carcinoma (NPC) presents a significant therapeutic challenge due to its aggressive nature and limited treatment options. Although morusin, a compound found in traditional Chinese medicines, exhibits significant tumor-inhibiting properties, its specific effects on NPC proliferation remain unclear. This study aims to elucidate the inhibitory effects of morusin on NPC survival and proliferation while exploring the underlying mechanisms through the utilization of network pharmacology, molecular docking, and experimental validation in vitro and in vivo. Network pharmacology analysis identified 117 potential targets of morusin against NPC, with 8 hub targets including AKT1, BCL2, CASP3, CTNNB1, ESR1, HSP90AA1, MMP9, STAT3, and the IL-17 signaling pathway. Further investigation of public data indicated that the expression levels of BLC2, CASP3, CTNNB1, HSP90AA1, and STAT3 in NPC tissue were significantly elevated compared to normal nasopharyngeal tissue. Docking studies exposed robust binding activity between morusin and key gene molecules. Additionally, biological assays demonstrated that morusin effectively inhibits NPC growth both in vivo and in vitro. Through a comprehensive investigation, this study identified the pharmacological mechanisms essential for morusin-induced inhibition of NPC growth by targeting multiple molecular targets and signaling pathways. These findings show the potential to contribute to the development of novel clinical agents for treating NPC.

Keywords: morusin, nasopharyngeal carcinoma; network pharmacology; therapeutic pathways..

Figure 1

The workflow chart of this study.

Figure 2

Identification of NPC-related targets and pathways. (A) 2913 NPC-associated targets, the red round rectangle and cyan-green round rectangles represent NPC and NPC-associated targets, respectively. (B) The 10 representative terms with the lowest p-value of biological processes (BPs), cellular components (CCs), and molecular functions (MFs) of the GO enrichment analysis (p < 0.05). (C) The 20 significantly enriched KEGG pathways of NPC-associated targets (p < 0.05).

Figure 3

Constitutional formula for morusin.

Figure 4

Identification of morusin-related targets and pathways. (A) 350 morusin-associated targets, the red round rectangle and cyan-green round rectangles represent morusin and morusin-associated targets, respectively. (B) Morusin-related targets were visualized by a Venn diagram, incorporating data from PharmMapper, CTD, SwissTargetPrediction, and TargetNet databases. (C) The 10 representative terms with the lowest p-value of biological processes (BPs), cellular components (CCs), and molecular functions (MFs) of the GO enrichment analysis (p < 0.05). (D) The 20 significantly enriched KEGG pathways of morusin-associated targets (p < 0.05). (E) Morusin-target-pathway network with morusin, pathways, and enriched targets represented as a green round rectangle, red V nodes, and blue circles, respectively.

Figure 5

Identification of morusin potential therapeutic pathways. (A) 117 common targets of morusin and NPC-associated targets were visualized by a Venn diagram. (B) 117 common targets of morusin and NPC-associated targets, the red round rectangle and cyan-green round rectangles represent morusin or NPC and morusin or NPC-associated targets, respectively. (C) The 18 significantly enriched GO pathway terms of 117 intersection targets. (D) The 20 significantly enriched KEGG pathway terms of morusin and NPC-associated targets (p < 0.05).

Figure 6

PPI network of morusin with the core targets in NPC setting. (A) The PPI network of morusin in NPC. (B) Ten hub targets were screened by Venn diagram. (C) The most significant module was obtained from MCODE analysis and the red circles represent hub targets.

Figure 7

Molecular docking results of morusin and hub targets. (A) Morusin-AKT1. (B) Morusin-CTNNB1. (C) Morusin-HSP90AA1. (D) Morusin-BCL2. (E) Morusin-STAT3. (F) Morusin-CASP3. (G) Morusin-ESR1. (H) Morusin-MMP9.

Figure 8

The core targets expression in different NPC cell types. (A, B) UMAP plot showing 10 clusters. (C) The core targets expression in different cell types.

Figure 9

Morusin inhibits cell viability in NPC cells and proliferation in the xenografts mouse model. (A, B) Effect of morusin on the viability of CNE1 and CNE2 cells. (C, D) Representative images and number of colonies formed by CNE1 and CNE2 cells. (E, F) The tumor volume of the xenografts treated with or without morusin (n=5). (G, H) The images of excised tumors in the two groups. * p < 0.05, comparison to the control group.