Akademik Veri Yönetim Sistemi
3rd International Cancer and Ion Channels Congress, İstanbul, Türkiye, 16 - 18 Eylül 2021, ss.71
Purpose: Cancer accounts for almost 8.2 million deaths every year worldwide. For this reason, cancer treatment is becoming increasingly important as a research issue and new drug discovery continues being a hot topic. Novel pharmaceutical products from natural active components have been increasing in recent years and used in the cancer treatment. In this respect, the anticancer effect of bornyl acetate (BA), which is one of the main components of Inula species, which is also endemic in our country, and its synergistic effect with 6- gingerol (6G) was investigated in MCF7 (breast cancer cell line) and breast cancer stem cells.
Materials and Methods: In the present study, cytotoxicity, apoptotic effect and cell morphology was revealed. Then, immunocytochemical examinations (Bax and Bcl-2, pro- and anti-apoptotic markers) were implemented. For this purpose, MCF7 cells and breast cancer stem cells isolated from MCF7 cells by MACS method which is a magnetic cell separation technique, were plated into 75 cm2 flasks in a ready-to-use medium supplemented with fetal bovine serum and penicillin-streptomycin in an incubator at 37 °C with 5% CO2. After sufficient confluency (70-80%) was achieved, the cells were removed from flasks with trypsin-EDTA and plated into 96 well-plates. After 24 h treatment, the cells were incubated with various concentrations of BA, 6G and BA/6G ranging from 0-600 μM. 24 h later, MTT cytotoxicity assay was applied to determine the inhibitory effect of BA and 6G for 24, 48 and 72 h. Furthermore, the cells were treated with IC50 of BA and 6G, and H&E staining was performed for the observation of cell morphology. Then, the effect of apoptosis by BA and 6G was investigated by acridine orange/ethidium bromide (AO/EB) staining, Annexin V/PI and immunocytochemistry staining.
Results: Our MTT results showed that the BA, 6G and BA/6G treatment has cytotoxic effect in a dose-dependent manner on MCF7 and breast cancer stem cells and the IC50 value was determined. In H&E staining, we observed apoptotic bodies, vacuolization, swelling, nuclear condensations and other cellular degenerations on the cells. Furthermore, the characteristics of apoptotic morphology such as cytoplasmic blebbing and shrinkage of the cells and nuclear condensation was showed using the method of AO/EB staining. In Annexin V/PI staining, the percentage of apoptotic cells was found about 40 % in both cell types at IC50 value for 24 h. Additionally, increased Bax and reduced Bcl-2 expression was observed in immunocytochemistry examination upon test compounds treatment at 24 h.
Conclusion: BA, 6G and BA/6G exhibited the cytotoxic and apoptotic effect and morphological changes on MCF7 human breast cancer cells and breast cancer stem cells. Therefore, the potency of BA and 6G for being anticancer agent ought to be examined in detail in future studies.