Antimycin A induced apoptosis in HCT-116 colorectal cancer cells through the up- and downregulation of multiple signaling pathways

Abstract

Colorectal cancer is the third most life-threatening cancer in the western countries. For the treatment, several chemotherapeutic drugs are using those that have severe side effects on the patient. So, finding alternative drugs is important. In the present research antimycin A was selected to evaluate the anticancer properties on the HCT-116 colorectal cancer cells. Antimycin A inhibited HCT-116 cells proliferation with the IC₅₀ value of 29 µg/mL concentration. As a long-term effect, HCT-116 cells were incubated with 10-40 µg/mL concentration of antimycin A for 7 days. No colony was observed in the treated wells. Apoptotic features in HCT-116 cells were observed in antimycin A treated cells after being stained with Hoechst 33342 dye. Apoptosis was further confirmed by FITC-annexin V/PI. Role of caspase-3 protein in the apoptosis process was also confirmed by the caspase-3 inhibitor. After treatment of HCT-116 cells with antimycin A, apoptotic related gene expression was checked by reverse transcription polymerase chain reaction. p53 and caspase-9 genes were upregulated consequently mitogen-activated protein kinases (MAPK), poly(ADP-Ribose) polymerase (PARP), and nuclear factor kappa B (NF-κB) genes were downregulated. Molecular docking simulation indicated significant binding affinity of antimycin A with the five proteins. The results indicated antimycin A would be a promising anticancer agent for further anticancer research.

Keywords: Antibiotic; Anticancer; Caspase activation; Gene expression; RT-PCR.