Exploring the mechanism of frankincense myrrh in the treatment of breast cancer based on data mining and network pharmacology
Frankincense is a resin extracted from the bark of the olive family plant Boswellia carterii Birdw. and the same plant Boswellia bhaurdajiana Birdw. It is pungent, warm, and bitter in taste, and is found in the heart, liver, and spleen meridians. It has the effects of promoting blood circulation, relieving pain, reducing swelling, and promoting muscle growth. Myrrh is a dry resin derived from the Euphora myrrh Engl. or Hemiphora molmol Engl. plants in the family Burseraceae. It has a mild nature, a pungent and bitter taste, and is suitable for the heart, liver, and spleen meridians. It has excellent effects in dispersing blood stasis, relieving pain, reducing swelling, and promoting muscle growth. The combination of frankincense and myrrh first appeared in the “frankincense pain relieving powder” in the “Guidelines for Diagnosis and Treatment”. In clinical practice, the two are often used in a complementary form, synergistically enhancing their efficacy, and are commonly used traditional Chinese medicine for promoting blood circulation and removing blood stasis.
At present, there are network pharmacology studies on frankincense and myrrh for arthritis, breast hyperplasia and cancer pain. These studies only directly reflect the biological process between the target and pathway of the compound, and there is no in-depth network pharmacology and related validation study on frankincense and myrrh for breast cancer treatment. Through consulting Chinese medicine books such as the “Dictionary of Tumor Formulas”, collecting clinical diagnosis and treatment data related to breast cancer, mining drug compatibility rules, core drugs and other association rules, this paper concluded that the “frankincense myrrh” drug pair has the highest confidence among the drugs used in Chinese medicine for breast cancer. Furthermore, network pharmacology and molecular docking studies were carried out on frankincense myrrh pairs to further clarify the active components and targets of frankincense myrrh, and analyze their molecular mechanisms in the treatment of breast cancer.

Through the analysis of the drug use rule in the target database by Apriori association rules, it was found that the highest confidence level was achieved in the simultaneous treatment of breast cancer with frankincense and myrrh. The higher the confidence level, the greater the possibility of the compatibility of myrrh and frankincense. CHAID, CRT, and QUEST are three commonly used decision tree prediction methods. CHAID uses multi-path splitting by default, which reduces the sample size in the nodes and leads to a decrease in the depth of the tree. CRT performs binary splitting (splitting each node into two child nodes). CHAID is designed to be used with classified/discrete targets, while CRT can be used for regression and classification. Compared to CART, CHAID has less confusion in the selection of split variables and split points. The QUEST node can provide a binary classification method for constructing decision trees, which is designed to reduce the processing time required for analyzing large CRT decision trees and to minimize the tendency of common bias towards predicting variables with multiple categories in classification tree methods. The calculation process is simpler and more effective than CRT, and the advantages of the three methods complement each other. The decision tree algorithm takes frankincense as the dependent variable, and the CHAID, CRT, and QUEST decision tree algorithms all screen for a higher possibility of compatibility between active ingredients and frankincense, indicating that frankincense and myrrh, as blood activating and stasis removing drugs, have significant implications for participating in the construction of the tree model formula. Frankincense was first recorded in the “Mingyi Bielu”. It has a warm nature and a mild taste, with a fragrant and flowing aroma. It is good at regulating qi and promoting blood circulation. The earliest record of myrrh dates back to the Song Dynasty’s “Kaibao Bencao”. It has a mild nature, thin qi, and a bitter taste, with a focus on removing blood stasis and regulating blood flow. The two are often used in synergy to enhance efficacy.
Further exploration of 53 active ingredients in frankincense and myrrh was conducted through the CytoNCA plugin in Cytoscape. Based on parameter degree ranking and clinical research literature, it was found that components such as 3-methoxyfuranone-9-en8-one, frankincense acid, and quercetin play important roles. Modern research has shown that the most characteristic and extensively studied component in frankincense is frankincense acid (such as frankincense acid), which belongs to the pentacyclic triterpenoid class and has anti proliferative, differentiation inducing, and apoptosis effects on tumor cells. In particular, acetyl frankincense acid has low toxicity and is a promising anti-tumor drug and tumor metastasis inhibitor. Furanones can also enhance the inhibitory activity of tumor cell proliferation to a certain extent. Quercetin is a polyphenolic substance found in various fruits and plants, which has antioxidant, anti-inflammatory, and pro apoptotic effects. Research has found that quercetin indirectly inhibits the expression of inflammatory factors interleukin-1 β (IL-1 β) and tumor necrosis factor alpha (TNF – α) by inhibiting the TLR-4/NF – κ B signaling pathway.
The GO function and KEGG pathway enrichment analysis of key targets were carried out to further explore the mechanism of frankincense myrrh in the treatment of breast cancer. GO contains three functional information: the biological process involved by genes, the cell components it has, and the molecular functions it plays. The results showed that frankincense myrrh may regulate PI3K Akt signaling pathway, Estrogen signaling pathway, MicroRNAs in by participating in 248 biological processes including positive/negative regulation of RNA polymerase II promoter transcription, transcription/DNA template, 60 cell components including nucleus, cytoplasm, and 94 molecular reactions including protein binding, poly (A) RNA binding, and enzyme binding. Cancer, MAPK signaling pathway and P53 signaling pathway play an anti-tumor role. Key genes are of great significance in the treatment of breast cancer with frankincense myrrh. The cytoHubba plug-in in Cytoscape was used to screen out the core genes with the highest degree value. According to the relevant references and the key pathways in KEGG enrichment analysis, two effective action pathways MicroRNAs in cancer and MAPK signal pathways were screened out. The core genes enriched in these two pathways are CREBBP, GRB2, HNRNPK, YWHAE, and YWHAG. The abnormal expression of MicroRNAs in cancer pathway and angiogenesis of breast cancer, tumor cell metastasis, invasion, and tumor drug resistance. The process is closely related, and miRNAs can be used as early diagnostic indicators for breast cancer, and hopefully become effective therapeutic drugs for breast cancer. Research has shown that myrrh steroids in myrrh may also exert anti-tumor effects by regulating the expression of miRNAs and affecting the activity of certain key target proteins. MAPK is a widely present serine/threonine protein kinase in cells. The MAPK signaling pathway participates in cell proliferation, apoptosis, autophagy, and regulates the expression of resistance related genes and proteins. Intervention in the MAPK signaling pathway can increase tumor sensitivity to chemotherapy drugs and reverse drug resistance. Studies have shown that β – elemene in myrrh also has good anti-tumor efficacy, and its anti proliferative effect on glioblastoma is achieved by activating the MAPK signaling pathway. YWHAE is closely related to the occurrence and development of tumors, mainly playing a role in pathways such as tumor cell cycle, growth and apoptosis, diffusion and migration, and signal transduction. CREBBP, as a transcriptional coactivator, can participate in a variety of cell functions and inhibit tumor. Its mutation can increase the incidence rate of ovarian cancer, follicular lymphoma and breast cancer. GRB2 is a widely expressed adaptor protein in cells, and its abnormal expression or activation is closely related to the occurrence of tumors. Studies have shown that it can serve as a molecular target for anti-tumor therapy, making GRB2 inhibitors a hot topic in the development of anti-tumor drugs. YWHAG is a tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activating protein gamma peptide (14-3-3 γ), which is involved in protein localization and transport, intracellular signaling, cell cycle regulation, and apoptosis. The HNRNPK gene can be distributed in the cytoplasm and/or nucleus of cells, and can interact with various proteins in different subcellular structures, participating in the regulation of the expression of multiple oncogenes and tumor suppressor genes in tumors.
In order to further verify the reliability of network pharmacology results, molecular docking technology was used to perform molecular docking on the core components and key targets separately. The results showed that the binding energies of the core components 3-methoxyfuran-guanosine-9-en-8-one, frankincense acid, and quercetin with the targets CREBBP, GRB2, HNRNPK, YWHAE, and YWHAG were all less than -5.0 kcal/mol, indicating their good binding activity. The binding energy of mastic acid and GRB2 is -9.1kcal/mol, indicating that they have strong binding activity, suggesting that GRB2 is an important target of mastic myrrh that may play a role in the treatment of breast cancer.
To sum up, this paper uses the network pharmacology method to build a “compound target disease” related network, and analyzes the interaction relationship between frankincense myrrh components, targets, and signal pathways as a whole. The molecular docking technology is used to verify the mechanism of frankincense and myrrh against breast cancer. It is speculated that frankincense and myrrh may act synergistically on CREBBP, GRB2, HNRNPK, YWHAE, YWHAG and other targets through 3-methoxyguanosyl-9-en-8-one, mastic acid, quercetin and other components, and through the MicroRNAs in cancer pathway MAPK signaling pathway is involved in cell proliferation, apoptosis, autophagy process and regulation of drug-resistant related genes, which shows the mechanism of frankincense and myrrh to play an anti-cancer role through multi-component, multi target, and multi pathway. Later, experimental studies can be carried out on frankincense myrrh pairs, to verify the results of this sub targeted docking virtual screening study, and to further explore the key mechanism of action, providing a new direction and basis for the research and development of drugs for breast cancer treatment.