Discussion
In the present study, we clearly demonstrated that frankincense, pine needle and geranium essential oils suppressed cell viability, proliferation and invasion in human BC cell line MCF-7. In addition, we determined that the frankincense, pine needle and geranium essential oils induced apoptosis, but did not affect cell cycle progression. The frankincense essential oil was also effective in inhibiting tumor growth and inducing tumor cell apoptosis in human BC mouse model. We demonstrated that frankincense, pine needle and geranium essential oils suppressed cell progression through the AMPK/mTOR pathway.
Essential oils, which are distilled from flowers, leaves, stems, the bark or roots of a specific plant, contained terpenes, aldehydes, esters, alcohols and other chemical molecules. It has been demonstrated that essential oils have an antibacterial and anti-inflammatory effect. Moreover, research has reported that essential oils also have a certain anticancer effect. Wu et al demonstrated that essential oils from Angelicae dahuricae and Inula japonica increased the sensitivity of BC cell line MCF-7/ADR to doxorubicin (21). Essential oils distilled from the leaves and flowers of Callistemon citrinus from the western Himalayas gave rise to the antiproliferative effect on human lung carcinoma cell line A549 and rat glioma C-6 cells via induction of apoptosis (22). Thymoquinone decreased proliferation and accelerated apoptosis in ID8-NGL (mouse ovarian cancer cells) tumors after 10 and 30 day-treatment (23). It has been observed thymoquinone mediated cell cycle arrest and apoptosis in BC and hepatocellular carcinoma (24,25). Boswellic acids (major components of frankincense) were reported to possess antitumor activity due to their cytostatic and pro-apoptotic properties in many human cancer cell lines containing meningioma (26), leukemia (27), hepatocellular carcinoma (28), melanoma, fibrosarcoma (29), colon (30) and prostate cancer (31-33). Moreover, the essential oil of frankincense inhibited proliferation and modulated apoptosis of human cancer cell lines both in vitro and in vivo (17,34,35). According to a study by Jeong et al, apoptosis, oxidative cell damage, induced by exposure to hydroxyl radical was inhibited by the extracts from pine needle (11). In the present study, we demonstrated that frankincense, pine needle and geranium essential oils reduced MCF-7 cell viability in a dose-dependent manner. Moreover, frankincense, pine needle and geranium essential oils strongly reduced colony size and colony forming capacity of MCF-7 cells. The treatment of frankincense, pine needle and geranium essential oils was responsible for the altered migratory and invasive phenotype of MCF-7 cells in vitro. Our results further indicated that the frankincense, pine needle and geranium essential oils induced apoptosis. However, the oils did not affect cell cycle progression. Furthermore, we observed that the essential oil of frankincense inhibited tumor growth and induced apoptosis in vivo. These results clearly revealed that frankincense, pine needle and geranium essential oils could play an important role in many biological functions of BC cells such as proliferation, invasion as well as apoptosis.
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Further analysis of molecular mechanisms for cancer cell progression may provide more data concerning novel molecular targets of frankincense, pine needle and geranium essential oil treatments for BC. AMPK is a central cellular energy-sensing system that constructively takes part in the interaction between metabolism and cancer progression by regulation of the mTOR pathway (36). The activation of AMPK directly phosphorylates and activates TSC2 by increasing its GAP activity and inhibiting mTOR signaling (37). The serine/threonine kinase mammalian target of rapamycin (mTOR) functions as a major regulator of cellular growth and survival, and resides in two multiprotein complexes, mTORC1 and mTORC2 (38). mTORC1 regulates phosphorylation of p70 S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E (eIF4E) binding protein 1 (4EBP1) (20,39). It was reported that aspirin decreased the viability and anchorage-independent growth of mutant PIK3CA BC cells through AMP-activated protein kinase (AMPK) activation and mTORC1 inhibition (40). Furthermore, in BC, 17-β-oestradiol (E2) directly activated AMPK through interaction of its α-subunit with estrogen receptors, implying its roles in cell proliferation (41). Knockdown of AMPK inhibited glucose metabolism and proliferation of TNBC cells (42). Based on the aforementioned, we analyzed the correlation of the frankincense, pine needle and geranium essential oils-mediated AMPK and mTOR signaling pathway. We revealed that these 3 essential oils notably regulated the activity of the AMPK/mTOR pathway in human BC cells MCF-7. These results demonstrated that frankincense, pine needle and geranium essential oil-modulated BC cell progression was, at least in part, AMPK/mTOR-dependent.
In conclusion, our results demonstrated that frankincense, pine needle and geranium essential oils have an antitumor effect, which could be mediated by the AMPK/mTOR pathway. The novelty of the present study was that the frankincense, pine needle and geranium essential oils may be a promising treatment for BC. However, the present study still had some shortcomings. To further confirm our present findings, similar experiments using the other BC cell lines should be performed. In addition, it is unclear whether the oils have an effect on BC patients. Thus, further investigation is warranted to analyze the effects and accurate mechanisms of frankincense, pine needle and geranium essential oils in BC.
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