Pyrazole Derivatives Induce Apoptosis via ROS Generation in the Triple Negative Breast Cancer Cells, MDA-MB-468.

Maryam Ashourpour,Ebrahim Saeedian Moghadam,Zahra Shahsavari,Mohsen Amini,Seyed Yousef Arman,Fatemeh Mostafavi Hosseini,Faranak Kazerouni

doi:10.31557/apjcp.2021.22.7.2079

Abstract

Background:Triple-negative breast cancer accounts for approximately 15–20% of all breast carcinomas and is associated with earlier age of onset, aggressive clinical course, and dismal prognosis. A series of 1,3-diaryl-5-(3,4,5-trimethoxyphenyl)-4,5-dihydro-1 H-Pyrazole and 1,3-diaryl-5- (3,4,5-trimethoxyphenyl)- 1 H-Pyrazole were evaluated for their anticancer activity against MDA-MB-468, human triple negative breast cancer cell line. Methods:The cytotoxic effects of Pyrazole derivatives on the growth of MDA-MB-468 and AGO1522 were determined using MTT assay. Annexin-V-FITC and PI staining were performed to detect apoptosis and cell cycle distribution using Flow cytometry. The level of Reactive oxygen species (ROS) formation and caspase 3 activity were determined accordingly. Results:Pyrazole derivatives induced a dose and time-dependent cell toxicity in MDA-MB-468 compared with untreated cells. The results showed that 3-(4-methoxyphenyl)-1-(p-tolyl)-5-(3,4,5-trimethoxyphenyl)-4,5-dihydro-1H-Pyrazole (3f) was the most active compound with IC50 values 14.97 μM and 6.45 μM compared with Paclitaxel with IC50 values 49.90 μM and 25.19 μM, after 24 and 48 hours, respectively. Upon treatment with 14.97 μM of 3f after 24 h, the compound induced cell cycle arrest in S phase. 3f provoked apoptosis was accompanied by the elevated level of ROS and increased caspase 3 activity in MDA-MB-468 cells compared with untreated cells. Conclusion:The overall results of the present study provided evidence for the cytotoxicity of compound 3f against MDA-MB-468 cells in comparison to reference standard, Paclitaxel. It proves that compound 3f can trigger apoptosis through ROS production and caspase 3 activation. These bring supportive data for future investigations that will lead to their use in cancer therapy.

Highlights

Breast cancer is one of the most common cancers among women and is a major reason for cancer death worldwide (Rakha et al, 2010)
The IC50 value obtained after 24 h treatment for MDA-MB-468 cancer cell and AGO1522 normal fibroblast cell line were recorded 14.97 and 28.74 μM respectively, indicating the effectiveness of 3f compound on inhibiting the growth of human triple negative breast cancer cells
Our results are in agreement with the literature that reported the apoptosis effect of Pyrazole derivatives on cancer cells (Inceler et al, 2013; Sebastian et al, 2016; Ananda et al, 2018; Nossier et al, 2018)

Summary

Introduction

Breast cancer is one of the most common cancers among women and is a major reason for cancer death worldwide (Rakha et al, 2010). Different Pyrazole derivatives including phenazone, metamizole, aminopyrine, phenylbutazone, sulfinpyrazone and oxyphenbutazone are readily available anti-inflammatory drugs (Prabhu et al, 2018). In this regard, Viagra , Celecoxib and Fipronil containing highly substituted Pyrazole ring have been commercialized as an effective ligand for estrogen receptors (Ananda et al, 2018). Sun et al, (2018) have reported the strong anti-tumor activity of these compounds for human lung cancer cell lines, A549. The present work aims to evaluate the anticancer properties of Pyrazole derivatives against MDA-MB-468 triple negative breast cancer cell line

Materials and Methods

Results

Findings

Discussion