To Explore the Mechanism of Muskone in the Treatment of Breast

Cancer based on Network Pharmacology

Haijun Chen

and Zhi Wen

Hunan University of Medicine, Huaihua 418000, China

Huaihua Hospital of Traditional Chinese Medicine, Huaihua 418000, China

Keywords: Muskone, Breast Cancer, Network Pharmacology, Mechanism of Action.

Abstract: To investigate the possible mechanism of Muskone in the treatment of Breast cancer through network

pharmacology. First, through Pubchem compound structure and Canonical SMILES, then use Swiss Target

Prediction and Targetnet database query targets of musk ketone, through Genecards and OMIM database

query targets for breast cancer disease, use the Target of muskone and breast cancer disease targets draw the

VENN diagram, using the STRING database and Cytosacpe3.7.1 software for network topology parameters

selection and muskone core targets for the treatment of breast cancer. Finally, the potential core targets were

analyzed through the DAVID platform for GO biological process and enrichment analysis of KEGG signaling

pathway. 135 targets of muskone and 1166 targets of breast cancer were obtained. There were 27 intersecting

targets between muskone and breast cancer. Cytosacpe3.7.1 software was used to analyze network topology

parameters, and 13 core targets of muskone in the treatment of breast cancer were obtained. A total of 78 GO

enrichment results were obtained. KEGG enrichment analysis revealed 9 major signaling pathways, including

cancer signaling pathway, prolactin signaling pathway, estrogen signaling pathway, etc. Muskone may be

used to treat breast cancer through multiple targets and different therapeutic approaches, which provides

reference for further study of pharmacodynamic substances and mechanism of action.

1 INTRODUCTION

At present, breast cancer has jumped to the first place

among female malignant tumors in China, especially

in urban areas. The mortality rate has increased by

96% compared with 30 years ago, breast cancer is

becoming the most threatening tumor to women

(Wang 2013). Breast cancer is A systemic disease

with strong clinical heterogeneity and complex

etiology. According to the origin or molecular

characteristics of cancer cells, it can be divided into

four molecular subtypes, namely luminal A/B, HER-

2 expression and triple-negative type. Meanwhile,

breast cancer is also the result of the combined effects

of environmental and genetic susceptibility factors

(Xin 2021). Western medicine has many methods to

treat breast cancer, such as surgical resection,

endocrine therapy, radiotherapy and chemotherapy,

but its side effects are obvious, increasing the

uncertainty of patients during the treatment. A large

https://orcid.org/0000-0002-2311-2748

https://orcid.org/0000-0001-6427-8034

number of studies have shown that traditional

Chinese medicine has great advantages in the

treatment of breast cancer. Traditional Chinese

medicine can be treated by dredging liver, regulating

qi and other syndromes, and it can significantly

relieve clinical symptoms and enhance patients'

confidence in the treatment of breast cancer.

Musk is the dry secretion from the mature male

musk capsule of forest musk deer, horse musk deer or

former musk deer, which has the functions of

activating the orifice and awakening the mind,

promoting blood circulation and menstruation,

reducing swelling and relieving pain (Li 1963).

Muskone is the main active component of musk, and

research results show that muskone has an important

function in malignant tumors (Qi 2020). However,

systematic and comprehensive studies on how

muskone plays a role in the treatment of breast cancer

from the cellular and molecular levels have rarely

been reported, and need to be further strengthened.

Chen, H. and Wen, Z.

To Explore the Mechanism of Muskone in the Treatment of Breast Cancer based on Network Pharmacology.

DOI: 10.5220/0011316000003444

In Proceedings of the 2nd Conference on Artiﬁcial Intelligence and Healthcare (CAIH 2021), pages 249-254

ISBN: 978-989-758-594-4

249

Based on this, this study explored the role and

mechanism of muskone in the treatment of breast

cancer through network pharmacology, as to provide

a reference for further study of the role of muskone in

the treatment of breast cancer.

2 METHODS AND RESULTS

2.1 Establishment of Chemical

Composition

Compound structure is obtained through the

Pubchem database (https://pubchem.ncbi.nlm.

nih.gov/) (see Fig.1) and Canonical SMILES

(CC1CCCCCCCCCCCCC (= NO) C1).

Figure 1: 3D structure of musk ketone.

2.2 Target Prediction of Muskone

The Canonical SMILES by Swiss Target Prediction

database of musk ketone gene targets, according to

the size of aim-listed Probability screen in the top 15

targets (see table 1), then Canonical SMILES input

Targetnet database (http://targetnet.scbdd.com/) of

muskone gene targets, selection targets which Prob >

0.The targets obtained from Swiss Target Prediction

database and Targetnet database were combined, and

the gene names were converted through Uniprot

(http://uniprot.org/) database, and the duplicates were

removed. A total of 135 related targets were obtained.

Table 1: Target of muskone.

Target Common name Probability

Cytochrome P450 19A1 CYP19A1 0.184

Carbonic anhydrase II CA2 0.112

Nuclear receptor subfamily 1 group I member 3 NR1I3 0.0940

Acyl coenzyme A: cholesterol acyltransferase CES1 0.0850

Carboxylesterase 2 CES2 0.0850

Carbonic anhydrase I CA1 0.0850

Carbonic anhydrase IV CA4 0.0760

Androgen Receptor AR 0.0490

P2X purinoceptor 7 P2RX7 0.0490

Poly [ADP-ribose] polymerase-1 PARP1 0.0490

Epoxide hydratase EPHX2 0.0490

Epoxide hydrolase 1 EPHX1 0.0490

Melatonin receptor 1A MTNR1A 0.0490

Melatonin receptor 1B MTNR1B 0.0490

Arachidonate 5-lipoxygenase ALOX5 0.0490

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2.3 Establishment of Breast Cancer

Related Targets

Enter the keyword Breast Cancer through Genecards

database (HTTP://www.genecards.org) and OMIM

database (https://www.omim.org/), search for gene

targets that have been reported and are related to

Breast Cancer. Gene targets with Score>20 were

selected from the data obtained from Genecards

database, according to the size of the Score value,

while the data obtained from OMIM database was not

selected because of the small number of data. Finally,

the data obtained from Genecards database and

OMIM database were combined, and the Uniprot

(http://uniprot.org/) database was used to convert

gene names and remove duplications. A total of 1166

disease targets were obtained.

2.4 Screening of Drug and Disease

Intersection Targets

Venn diagrams of drug and disease targets are plotted

(see Fig.2), 135 muskone targets with 1166 targets for

breast cancer disease input Venn platform

(https://bioinfogp.cnb.csic.es/tools/venny/). A total of

27 potential targets of muskone active components

against breast cancer were obtained: CYP19A1, AR,

PARP1, ABCB1, AHR, CA9, CASP9, CDK1, CDK2,

CDK4, CYP17A1, CYP1A2, CYP2D6, etc.

Figure 2: VENN diagram of the intersection target of

muskone and breast cancer target.

2.5 Screening of PPI Protein

Interaction Network and Core

Targets

The PPI network map was obtained by STING

database (https://string-db.org/) for 27 potential

targets of muskone active ingredients against breast

cancer. Import all relevant data directly obtained in

STING network database into Cytoscape3.7.1

software. Each node represents a target, and the edge

represents the interaction between the two targets.

The more connected node lines, the more critical the

target is, and the key proteins are CYP19A1, AR,

PARP1, ABCB1, AHR, CA9, CASP9, CDK1, CDK2,

CDK4, CYP17A1, CYP1A2, CYP2D6, DNMT1, and

ESR1 (see Fig.3). The muskone active substances

obtained in STING database and 27 potential action

targets effective against breast cancer were imported

into Cytoscape3.7.1 software to get the potential core

target, and the median of Degree value was 9.48,

median of Betweenness Centrality value was 0.0078,

and median of So-called Centrality value was 0.604.

Degree value, Betweenness Centrality, Closeness

Centrality values were greater than the value of

target, the target can be thought of as potential core

targets 13 muskone in the treatment of breast cancer,

such as ESR1, HSP90AA1, AR, that the above targets

may is the core of muskone is used to treat breast

cancer gene (see Table 2).

Figure 3: Network diagram of PPI protein interaction.

Table 2: Topological parameters of potential core targets of

muskone in the treatment of breast cancer.

Name

Betweenness

Centrality

Closeness

Centrality

Degree

ESR1 0.281 0.930 24

HSP90A

0.102 0.761 18

AR 0.0620 0.740 17

PTGS2 0.0700 0.702 15

SIRT1 0.0140 0.671 13

RELA 0.0450 0.650 12

CDK4 0.0100 0.650 12

DNMT1 0.0170 0.650 12

PARP1 0.0130 0.651 12

AHR 0.0360 0.630 11

PGR 0.0140 0.629 11

ESR2 0.0280 0.620 10

CYP19

0.0120 0.600 10

To Explore the Mechanism of Muskone in the Treatment of Breast Cancer based on Network Pharmacology

251

2.6 GO Biological Process Analysis

13 potential core targets were imported into DAVID

database for GO enrichment analysis. A total of 78

results were obtained from GO enrichment analysis.

There were 44 Biological processes (BP), 27

Molecular functions (MF) and 7 Cellular components

(CC). The top biological functions included core

promoter sequence specific DNA binding, enzyme

binding, protein binding, regulation of human

transcription binding factor activity complex,

positive and negative regulation of human cell

function growth, transcriptional regulation process,

positive regulation process of NF-κB transcription

factor activity, DNA binding and other processes. The

results with P value less than 0.05 were screened out

from the obtained data to draw a bar chart (see Fig.4).

The color of the column represents the GO biological

process information entry. The column height from

low to high means that the P value decreases from

large to small. The smaller the P value, the higher the

significance. This suggests that muskone may play a

therapeutic role in the treatment of breast cancer by

regulating these biological processes.

A. molecular function

B. cell component

C. biological process

Figure 4: GO enrichment analysis of muskone in the

treatment of breast cancer.

2.7 Enrichment Analysis of KEGG

Pathway

13 potential core targets were imported into DAVID

database for Enrichment analysis of KEGG pathway.

Results show that There were 9 main enrichment

pathways of key target genes in the treatment of

breast cancer by muskone. Five key signaling

pathways, namely pathways in cancer, Prolactin

signaling Pathway, Small cell Lung cancer, prostate

cancer, and estrogen signaling Pathway, are used to

play the anti-cancer role (see Table 3). The signal

conduction pathway with P value less than 0.05 was

taken as the bar chart. This is the main enrichment

pathway of key target genes in the mechanism of

action of muskone in the treatment of breast cancer,

indicating that muskone may treat breast cancer

through cancer transduction pathway, prolactin

signaling pathway, estrogen signaling pathway (See

Fig.5). In the figure, the height of the column

represents p-value, and the column height from low

to high represents the P value from large to small, and

the smaller the P value is, the stronger the

significance is.

Table 3: Pathway enrichment analysis table.

Term Count P-Value

Pathways in cancer 5 0.00250

Prolactin signaling pathway 3 0.00540

Small cell lung cancer 3 0.00770

Prostate cancer 3 0.00830

Estrogen signaling pathway 3 0.0100

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Figure 5: Enrichment analysis of KEGG metabolic pathway

in the treatment of breast cancer with muskone.

3 DISCUSSION

Traditional Chinese medicine thinks, cancer etiology

and pathogenesis of blood stasis block, musk can

begin to understand wake up, invigorate the

circulation of menstruation, acetanilide

detumescence, conforms to the principle of TCM

anticancer treatment, has antitumor effect, the study

also showed that musk can inhibit the growth of

cancer cells proliferation, muskone as main

ingredients, inhibit breast cancer tissue fibroblast

growth factor, bFGF and VEGF expression antitumor

the formation of new blood vessels, thereby

potentially anticancer(Meng 1998). However, there

are few reports on the molecular mechanism of

musk's anti-breast cancer and anti-drug resistance and

its therapeutic effect on breast cancer. Therefore, to

explore the molecular mechanism of musk ketone in

the treatment of breast cancer is conducive to further

research, promote the development of Traditional

Chinese medicine, and enhance the cultural

confidence of traditional Chinese medicine.

Researchers in this study by using the way of

network pharmacology, by constructing PPI network

discovery ESR1, HSP90AA1, AR, PTGS2, SIRT1,

RELA, CDK4, DNMT1, PARP1, AHR, PGR, ESR2,

CYP19A1 as the core target of muskone breast cancer

treatment, illustrates the muskone can through a

variety of target gene therapy of breast cancer,

consistent with modern research.

ESR can specifically bind with estrogen and

participate in the occurrence and development of

breast cancer by activating SRC-Ras-PI3K-Akt and

MAPK/ERK. AR can combine with target genes and

activate them, thus activating the downstream cell

proliferation signaling pathway and participating in

the proliferation of mammary epithelial cells (Filardo

2002).

As a molecular protein family, heat shock protein

family is involved in the whole process of tumor

occurrence and development. HSP90α protein is

increased in breast cancer patients. Studies have

shown that HSP90 encoded by HSP90AA1 gene can

inhibit apoptosis, regulate cell division and promote

angiogenesis (Chatterjee 1978). Through TCGA data

analysis found that HSP90AA1 mRNA was highly

expressed in breast cancer tissues and was related to

patient survival (Jia 2020). PGR is a target of

endocrine therapy for breast cancer and has good

reference value for prognosis of breast cancer. Aryl

hydrocarbon receptor (AHR) is a ligand-dependent

activated transcription factor, which is associated

with the occurrence of breast cancer and can regulate

the proliferation and apoptosis of tumor cells.

Abnormal expression of PARP1 gene is closely

related to triple-negative breast cancer (TNBC), and

PARP1 may be involved in the treatment of breast

cancer by inhibiting angiogenesis (Feng 2020).

DNMT 1 regulates the occurrence and development

of breast cancer by regulating the migration and

proliferation of breast cancer cells (Li 2020). Cyclin-

dependent kinase CDK4 is a key regulator of cell

cycle. In estrogen receptor positive (ER+) breast

cancer, CDK4 is overexpressed and cell proliferation

is uncontrolled. RELA is a member of NF-κB family.

NF-κB gene in breast cancer cells affects the invasion

and migration ability of breast cancer cells by

inhibiting the epithelial-mesenchymal transformation

of tumor cells (Zhao 2019). Sirtuins family proteins

are a class of NAD+ dependent deacetylases involved

in the development and Multidrug resistance (MDR)

of breast cancer (Li 2021). PTGS 2 gene variation is

associated with breast cancer susceptibility (Uwe

2006, Laure 2010). The expression of aromatase gene

(CYP19A1) has a certain effect on the biological

behavior of tumor cells (Luo 2014).

In order to predict and muskone key targets for

therapy of breast cancer in gene function and the role

of signaling pathways, this study analyzes the GO

biological function of enrichment, found that the core

PPI gene may be through the core promoter sequence

specific dna-binding proteins, enzyme combination,

combined with activity, as well as the combination of

human transcription factor complex regulation, to the

human body cell function positive negative

regulation of growth, transcriptional regulation, the

nf-kappa B the activity of transcription factors are

regulation and DNA binding have the effect of breast

cancer.

Through KEGG enrichment analysis, the results

showed that the target genes of breast cancer treated

by muskone mainly involved pathways in cancer,

Prolactin signaling pathway, Small cell lung cancer,

To Explore the Mechanism of Muskone in the Treatment of Breast Cancer based on Network Pharmacology

253

prostate cancer, estrogen signaling pathway. These

pathways are closely related to the formation and

development of cancer. The prolactin signal

transduction pathway and lactin signal transduction

pathway mainly control and treat breast cancer by

regulating human hormones. This study suggests that

muskone may be mainly used in the treatment of

breast cancer through the regulation of human

transcription binding factor activity complex,

positive regulation process of NF-κB transcription

factor activity, cancer pathway, prolactin pathway

and estrogen signal transduction pathway.

This study adopts the network pharmacology

method to predict the mechanism of muskone in the

treatment of breast cancer, which involves a variety

of biological processes and multiple pathways,

reflecting the multi-target-multi-pathway action

characteristics of Chinese medicine components. At

the same time, it was found that multiple targets

interact to regulate a signal pathway and affect the

biological response, reflecting the characteristics of

muskone as a Chinese medicine ingredient in the

treatment of breast cancer. However, the

experimental results still need further verification and

further development. With the deepening of the

research on the pharmacological action and

mechanism of natural musk and muskone, the

medicinal value and market prospect of musk will be

better displayed.

4 CONCLUSIONS

Based on the results and discussions presented above,

the conclusions are obtained as below:

(1) It is shown that 135 targets of muskone and

1166 targets of breast cancer were obtained. There

were 27 intersecting targets between muskone and

breast cancer. Cytosacpe3.7.1 software was used to

analyze network topology parameters, and 13 core

targets of muskone in the treatment of breast cancer

were obtained. A total of 78 GO enrichment results

were obtained. KEGG enrichment analysis revealed

9 major signaling pathways, including cancer

signaling pathway, prolactin signaling pathway,

estrogen signaling pathway.

(2) Muskone may be used to treat breast cancer

through multiple targets and different therapeutic

approaches, which provides reference for further

study of pharmacodynamic substances and

mechanism of action.

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