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  • CH5138303 br Experimental Procedures br Author

    2021-02-20


    Experimental Procedures
    Author Contributions
    Acknowledgments
    Introduction Breast cancer is one of the most common malignancies for women worldwide, representing approximately 22% of women’s malignancies that pose a threat to women’s health [[1], [2], [3], [4]]. The morbidity rate of breast cancer, which used to remain relatively low in developing countries like China, has globally increased by about tenfold over the decade. For instance, in China, there is an annual rise of 2.7% in morbidity rate. Due to the complicated pathogenesis of breast cancer, the existing treatments work effectively in prolonging the survival of patients, but still have some limitations [[5], [6], [7], [8], [9]]. In China, an increasing number of traditional Chinese medicines have been discovered to possess anti-inflammatory and tumor-suppressing abilities, which may open up brand new vistas for traditional Chinese medicines [10,11]. The family Bufonidae, in the order Anura (frogs and toads), is widely distributed in China, with the two common species Bufo gargarizans and Duttaphrynus melanostictus. ChanSu and ChanTui (the molting skin of toad), treasures of Chinese medicines, contain various chemicals and can be clinically applied to reduce inflammation, relieve pains, strengthen the heart, treat cancers, etc [[12], [13], [14], [15]]. Currently, Germany, Japan and China have used toads to manufacture medicines for clinical treatment. For example, bufalin, which is isolated from ChanSu, could activate pro-caspase3/8/9, up-regulate Bax/Bcl-2 and down-regulate inhibitors of apoptosis proteins, thereby causing mitochondrial energy imbalance and leading to the apoptosis of CH5138303 cancer cells [16,17]. Further, arenobufagin, another compound from ChanSu, could induce apoptosis of colon cancer cells via Bax-PGAM5LDrpl complex [18]. Thus, previous studies have demonstrated that the compounds extracted from ChanSu could lead to the death of cancer cells, inhibit the proliferation of tumor, and perform other anti-tumor functions. Telocinobufagin (C24H34O5, Molar mass 402.52, Fig. 1), which is also isolated and purified from ChanSu, has various pharmacological properties such as strengthening cardiac muscles, fighting against microbes, boosting the immune system, etc [[19], [20], [21]]. However, no research or reports have been found on the role of telocinobufagin in breast cancer.
    Methods and materials
    Results
    Discussion The clinical application of ChanSu in China dates back 1400 years, and was documented in The Introduction to Medicine Properties, an ancient medical book written by Tang Zhen-quan. Historically, ChanSu was used to cure cancers and improve the immune system [37,38]. Despite this, only a few studies concentrated on the role and molecular mechanism of ChanSu in cancer treatment. The function and mechanisms of telocinobufagin, a chemical extracted from ChanSu, have been reported only by several groups and not yet been well clarified. The results of Liang et al. revealed that telocinobufagin could induce apoptosis and, therefore, obviously suppress the cell viability of HCT116 and SW480, which might be related to the activation of P53-mediated Bax pathway and the blockage of IAP pathway in colon cancer cells [19]. Meanwhile, an activity value of 2.4 mM of telocinobufagin has been reported on human HepG2 cells after 24 h using MTT assay (Substance SID: 103601053, BioAssay AID: 773796) by PubChem Com. Similarly, an activity value of 3 mM of telocinobufagin on A549 cells has also been documented by PubChem Com (Substance SID: 103601053, BioAssay AID: 773795, https://pubchem.ncbi.nlm.nih.gov/compound/259991#section=Biological-Test-Results). The effect of telocinobufagin on breast cancer has only been reported once. Zhang et al. revealed that telocinobufagin exposed noteworthy cytotoxic activities against breast cancer (MCF-7) cell lines [39]. However, the related genes and pathways of telocinobufagin in breast cancer have never been reported. In this study, the in silico approach was employed to investigate the molecular mechanism of telocinobufagin in breast cancer. Moreover, taking advantage of public databases, we conducted a standardized analysis of studies regarding the role of telocinobufagin in curing breast cancer. Finally, we acquired expression profiles of 1251 differentially expressed genes, which were considered as reliable examples of gene alteration after the breast cancer cells were treated with telocinobufagin.