IInvestigate the effects of linearol on the endoplasmic reticulum markers by using the endoplasmic reticulum stress generated model of human breast cancer cells (MCF-7).

Cancer is a multi-stage process in which functional and behavioral differences occurs in the cell and cell escapes from the control of the immune system, which consequently leads to form metastases in the tissue. Breast cancer is characterized by uncontrolled growth of malignant cells in the breast epithelium tissue. The disease can be effective in both sexes. Endoplasmic reticulum (ER) is an important organelle species required for normal cellular processes such as regulation of protein synthesis, folding of proteins, calcium storage, control of cholesterol and lipid-membrane biosynthesis. Accumulation of misfolded proteins in the ER creates the UPR response as a protective response that reduces the synthesis of new-on-protein, facilitates protein folding, and increases proteosomal degradation of misfolded proteins. In this study, the endoplasmic reticulum stress was induced by thapsigargin in human breast cancer (MCF-7) cells and the cytotoxicity of acetone extract of Sideritis akmanii (S. akmanii) as a linearol source in stressed cells was determined by 3- (4,5-dimethyltriazol- diphenyltetrazolium bromide (MTT) method. Following doses (1000, 750, 500, 250, 100, 50, 25 μg / ml) of S. akmanii were employed and 1% DMSO was used as a solvent control. The possible effect of the MTT method on the MCF-7 cells by the S. akmanii was studied with a microplate reader at 540 nm and the results were expressed as a percentage. Control was accepted as 100% live and was determined to be 38,851 μg / mL by the proband analysis of lecithin (LD50) of SAE. In our study, the effect of the acetone extract of S. akmanii as a linearol source in MCF-7 cells with normal and ER stress was further studied by the expression level of ATF-4, AFT-6, PERK, Grp78, TNFα, IFN-γ, IL-6, IL-8 and IL-12 genes by real time PCR. According to the results of our study, the expression levels of Grp78 gene in the SAE50 and SAE50 + T treatment groups, which were involved in UPR response in ER stress, were found to be higher than control levels. It was observed that the PERK gene involved in UPR response in ER stress was suppressed in all application groups but increased in SAE100 + T administration compared to control. It was also found that ATF-6, which is involved in UPR response in ER stress and in control of Grp78, increases expression in SAE50 and SAE50 + T treatment groups like Grp78. An increase in IL-6 gene expression was observed in all treatment groups compared to that of control. IL-8 gene expression levels was suppressed in SAE10, SAE10 + T and SAE50 + T applications and increased in other treatment groups. Similar expression was observed in IL-12 and TNFα genes whereas these were suppressed in all other treatment groups. The IFN-γ gene showed the same effect as the control at the LD50 concentration obtained by the cytotoxicity test, but suppressed in the other treatment groups. The highest DNA damage score was found as 100 μg / mL (94.00 ± 5.29) in the Thapsigargin treated group. it was concluded that S. akmanii has genotoxic and cytotoxic effects on MCF-7 cells. Key Words: MCF-7, ER stress, Comet Assay, Sideritis akmanii, Real Time PCR, Genotoxicity, Cytotoxicity, Endemic plant