Cyclophosphamide (CTX), also known as cytophosphane among other, is a medication used as chemotherapy and to suppress the immune system. The PI3K/AKT/mTOR pathway is involved in the regulation of diverse cellular functions, including cell growth, protein synthesis, cell cycle regulation, glucose metabolism, and motility. In our study eight weeks old C57BL/6 female mice were divided into 3 groups as control (C), sham (S) and experimental group. The experimental group has been established with CTX treatment. No treatment was applied to the C group. The S group were given an equal amount of saline. CTX was administered intraperitoneally one every 2 days for 3 weeks; the first dose was 70 mg/kg, the ongoing doses were 30 mg/kg. At the end of 3 weeks mice were sacrificed and kidneys were taken for investigation. In order to show the effect of cyclophosphamide in kidney tissue, the tissues were stained via indirect immunohistochemistry with PI3K, AKT and mTOR primary antibodies. In our study, PI3K, AKT and mTOR expression levels were found to be significantly decreased in CTX-mediated mechanisms indicating that the mechanisms of CTX might involve in the inhibition of PI3K/AKT/mTOR signaling pathway.

Phytotherapy has been used for many years due to anticancer and anti-proliferative effects. In this study our purpose was to show the anti-proliferative and apoptotic effects of oleocanthal and pinus pinaster on MCF-7, MDA-MB-231, 67NR and 4T1 breast cancer cell lines. Biological effects of these plants were researched via morphology, MTT assay for cytotoxicity (IC50), immunocytochemical procedure for oxidative stress (eNOS), angiogenesis (VEGF) and TUNEL method for apoptosis. Statistical analysis was performed with the H-score. Oleocanthal and Pinus pinaster extracts showed significant dose and time-dependent inhibition of growth of breast cancer cells. A significant increase in iNOS staining was observed while a decrease in VEGF staining was observed after extract application at IC50 dose. The results showed a significant increase in apoptosis in extract-treated breast cancer cell lines. We showed that the toxic effect of Oleocanthal and Pinus pinaster extracts created by oxidative stress mechanisms. The increase of oxidative stress in breast cancer cells caused the inhibition of cell proliferation and apoptosis. Our results indicated that these plants suggest potential agents in breast cancer treatment. Future studies will focus on the identification of the molecules responsible for anti-cancer activity of these substances in order to improve the quality of life of the patients.

Inula viscosa, Rubus caesius and Viscum album known as medicinal plants which have therapeutic properties in treatment of various diseases and particularly cancer. In this study, we aimed to show their anti-proliferative and apoptotic effects on MCF-7, MDA-MB-231, 67NR and 4T1 breast cancer cell lines. The cell lines were cultured to determine the cell growth using the MTT assay. Cells were cultured to evaluate for eNOS, VEGF and apoptosis by immunocytochemical staining and TUNEL assay. Statistical analysis was performed with the H-score. Inula viscosa, Rubus caesius and Viscum album extracts were found to significantly inhibit the growth of breast cancer cell lines in dose and time-dependent manner. eNOS staining was significantly increased and VEGF was decreased after extract application at IC50 dose. Similarly, results indicated that the treated breast cancer cell lines exhibited a marked increase in apoptosis. We showed that the toxic effect of Inula viscosa, Rubus caesius and Viscum album extracts created by oxidative stress and apoptotic mechanisms. Our findings suggested that due to their anticancer potential, these plants may be used as an alternative treatment for the management of cancer. Therefore future studies will focus on the identification of the molecules responsible for the anticancer activity.