Investigation of anti-cancer activities of PhTAD-substituted dihydropyrrole derivatives compounds

In the fight against breast cancer and all other types of cancer, determining the molecular pathways that are effective in the development and spread of cancer, the discovery of new active substances that may exhibit an anticarcinogenic effect or the determination of the effectiveness of some chemicals are very important in terms of creating new and more effective treatment options. For this purpose, in our study, new derivatives of biologically active bicyclic dihydropyrol compounds formed by the addition of N-phenyltriazolinedione (PhTAD) is aimed to determine the relationship between anticancerogenic activities and gene expression changes in MCF-7 (breast cancer) and MCF-12A (normal breast epithelium) cell lines. In our study, in the MCF-7 and MCF-12A cell lines, cytotoxic effects and IC50 values of sixteen different PhTAD-derived dihydropyrol compounds by MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) method, their effects on cell proliferation were determined by real-time cell analysis system (RTCA). In addition, in the cell lines of these compounds apoptotic cell death proteins P53, BAX, BCL2, PARP and proteins that play an important role in protein with related signaling pathways HSP27, HSP40, HSP60, HSP70, hsp90α and GRP78, and GRP94's which is the main regulatory proteins in the endoplasmic reticulum (ER) stress gene expression levels were investigated by real-time polymerase chain reaction (RT-PCR) method. As a result of our study, it was observed that compounds of different doses (1.56-100 µM) applied to the MCF-7 cell line exhibited moderate and strong cytotoxic and antiproliferative activity by both MTT and RTCA method. Also, many of these compounds have been found to have effects that reduce p53 gene expression. However, it has been concluded that a significant portion of these compounds increase the BAX / BCL2 ratio and decrease the level of PARP1 gene expression, leading to apoptosis independent of p53. In addition, these compounds, especially HSP90, which is considered as an important strategic target in breast cancer, other important HSPs have been shown to have a reducing effect on gene expressions and also have a suppressive role on GRP78 and GRP94 gene expression, which contribute to tumorization by re-folding proteins in ER stress. In conclusion, the effects of PhTAD-derived dihydropyrol compounds on MCF-7 cell line to stimulate apoptotic genes and the presence of suppressive effects on the expression of genes in the regulation of ER stress with apoptosis-related signaling pathways indicates that these compounds increase apoptotic activity. Considering these effects, it is concluded that these compounds can be used as anticancer agents.