Usama bin Naeem¹, Muhammad Adil Rasheed^1*, Muhammad Ashraf¹, Muhammad Yasir Zahoor²

¹Department of Pharmacology and Toxicology, Faculty of Biosciences, University of Veterinary and Animal Sciences Lahore, Pakistan

²Insititue of Biochemistry and Biotechnology, Faculty of Biosciences, University of Veterinary and Animal Sciences Lahore, Pakistan

Abstract

One of the most dominant diseases in the world, particularly among women, is breast cancer. Breast cancer has tumor suppressor genes called CHEK2 and TP53. When there is a mutation in CHEK2 and TP53 genes there are more chances of breast cancer. This study aimed to investigate the already prepared and characterized nanoparticles loaded with Chitosan for Cell death, Mitochondrial Membrane and cell cycle arrest estimated through Flow Cytometry and gene expression analysis of CHEK2 and TP53 genes by real-time PCR. The Livak method was used to evaluate the results. The mean (± S.D) comparison between the control and target genes were used to calculate gene expression. Results showed that Ivermectin and Tamoxifen NPs (B+C) represented 34.8% cell death that is better than other combinations with propidium iodide stain while with Acridine orange stain Tamoxifen+Ivermectin (A+B) combination showed the remarkable and maximum of the all cell cycle arrest with value of 69.7% cell arrest at G0/G1 phase, 7.11% of cell arrest at S Phase and 7.05% of G2/M Phase arrest. It was demonstrated that the expression levels of CHEK2 and TP53 genes were significantly increased (P<0.001) in Ivermectin+Tamoxifen NPs (B+C) compared with control groups. It is concluded that Tamoxifen nanoparticles with Ivermectin showed strong anti-proliferative activity against breast cancer cells. The expression levels of nanoparticles containing Tamoxifen were significantly increased compared to the other treatments and control groups (P<0.001). Gene expression change with change in dose concentrations.

Keywords: Breast cancer, Apoptosis, Cell cycle arrest, Pharmacogenomic, Gene expression