Multidimensional analysis to elucidate the possible mechanism of bone metastasis in breast cancer.

BACKGROUND: Breast cancer (BC) patients tend to suffer from distant metastasis, especially bone metastasis. METHODS: All the analysis based on open-accessed data was performed in R software, dependent on multiple algorithms and packages. The RNA levels of specific genes were detected using quantitative Real-time PCR as a method of detecting the RNA levels. To assess the ability of BC cells to proliferate, we utilized the CCK8 test, colony formation, and the 5-Ethynyl-20-deoxyuridine assay. BC cells were evaluated for invasion and migration by using Transwell assays and wound healing assays. RESULTS: In our study, we identified the molecules involved in BC bone metastasis based on the data from multiple BC cohorts. Then, we comprehensively investigated the effect pattern and underlying biological role of these molecules. We found that in the identified molecules, the EMP1, ACKR3, ITGA10, MMP13, COL11A1, and THY1 were significantly correlated with patient prognosis and mainly expressed in CAFs. Therefore, we explored the CAFs in the BC microenvironment. Results showed that CAFs could activate multiple carcinogenic pathways and most of these pathways play an important role in cancer metastasis. Meanwhile, we noticed the interaction between CAFs and malignant, endothelial, and M2 macrophage cells. Moreover, we found that CAFs could induce the remodeling of the BC microenvironment and promote the malignant behavior of BC cells. Then, we identified MMP13 for further analysis. It was found that MMP13 can enhance the malignant phenotype of BC cells. Meanwhile, biological enrichment and immune infiltration analysis were conducted to present the effect pattern of MMP13 in BC. CONCLUSIONS: Our result can improve the understanding of researchers on the underlying mechanisms of BC bone metastasis.

Root extract of Prunella vulgaris inhibits in vitro and in vivo carcinogenesis in MCF-5 human breast carcinoma via suppression of angiogenesis, induction of apoptosis, cell cycle arrest and modulation of PI3K/AKT signalling pathway.

PURPOSE: In this study we examined the anticancer effects of methanolic root extract of Prunella Vulgaris (PVE) against the MCF-5 breast cancer (BC) cell line along with its mode of action. METHODS: The proliferation rate of the MCF-5 cells was assessed by MTT assay. Apoptosis was confirmed by acridine orange (AO)/ethidium bromide (EB) and annexin V/propidium iodide (PI) staining. DNA damage was checked by comet assay. Cell cycle analysis was performed by flow cytometry. Protein expression was determined by western blotting. In vivo evaluation of the extract was carried out in xenografted tumor mice models. RESULTS: PVE inhibited the growth of the MCF-5 cells and exhibited an IC50 value of 25 µg/ml. The investigation of underlying mechanism revealed that PVE triggered apoptotic cell death of the MCF-5 cells which was also associated with enhancement of the expression of Bax and decrease in the expression of Bcl-2. PVE also caused arrest of the cells in the G2/M phase of the cell cycle and also exerted the anti-angiogenic effects. In vivo evaluation of PVE showed that it could inhibit the tumor weight and volume, suggestive of the anticancer potential of PVE. CONCLUSION: The root extract of Prunella vulgaris in this study was shown to exert potent anticancer effects in MCF-7 human BC cells both in vitro and in vivo, accompanied with apoptosis induction, inhibition of angiogenesis, cell cycle arrest, and modulation of PI3K/AKT signaling pathway.