Cumulative effects of ciprofloxacin and pilocarpine on cytotoxicity and G0 phase arrest in hepatoma-derived Hep G2 cell line

Abstract

Objectives Uncontrolled cell proliferation was caused by multiple deficient pathways that inhibition of one pathway may result to activate an alternative pathway. Therefore, combination of drugs which targeted multiple pathways could be beneficial to overcome drug resistance. Ciprofloxacin (CPF) cytotoxicity was widely investigated on cancer cell lines, and results revealed hepatoma‐derived Hep G2 cells are relatively resistant. So, this study aimed to increase CPF cytotoxicity by rational design of a supplement which targeted Ca2+ homoeostasis as major hub in unchecked proliferation. Methods Cells were treated by CPF and/or pilocarpine (PILO), and cell cycle distribution, caspases activity and regulatory proteins were evaluated. Key findings MTT and flow cytometry analysis confirmed administration of CPF + PILO causes more cytotoxicity. CPF‐exposed cells accumulated in S phase due to DNA damages while PILO + CPF imposed G0 stage arrest through cyclin D1 and P‐Akt downregulation. Caspase 8 was activated in cells treated by CPF but accompaniment of PILO with CPF led to activation of caspase 9, 8 and 3 and ROS overproduction. Conclusions Ciprofloxacin imposed mitochondrial‐independent apoptosis while PILO + CPF caused mitochondrial‐dependent and independent apoptosis simultaneously. Consequently, coadministration of PILO and CPF causes intense cytotoxic effects through targeting the mitochondria, DNA gyrase enzyme and other unknown mechanisms.

N-Acylated Ciprofloxacin Derivatives: Synthesis and In Vitro Biological Evaluation as Antibacterial and Anticancer Agents

“…What is more, the anticancer activities of fluoroquinolones are an effect of restraining of eukaryotic topoisomerase IIα, the analogue of DNA gyrase. It results in the formation of double-strand breaks in nucleic acids and incomplete DNA synthesis, which lead to S-phase arrest in a cancer cell . Since the affinity of CP to the eukaryotic enzyme is weaker than to the prokaryotic homologue, an anticancer treatment requires higher drug dosages in comparison with quantities used in bacterial infections.…”

“…Cancerous cells exhibited different susceptibilities to this chemotherapeutic. − Hepatocellular cancer cells (Hep G2) remained quite resistant, whereas colon carcinomas (SW-403) were found to be more sensitive . It has been estimated recently that the fluoroquinolone derivative induces oxidative stress in hepatoma-derived cells, increases activation of caspases involved in different pathways of cell death, and imposes double-strand breakages in the cellular DNA . Moreover, CP triggers apoptosis of human triple-negative breast cancer (MDA-MB-231) cells by leading to a loss of the mitochondrial transmembrane potential and stimulation of the cell cycle arrest at the S-phase .…”

“…It results in the formation of double-strand breaks in nucleic acids and incomplete DNA synthesis, which lead to S-phase arrest in a cancer cell. 37 Since the affinity of CP to the eukaryotic enzyme is weaker than to the prokaryotic homologue, an anticancer treatment requires higher drug dosages in comparison with quantities used in bacterial infections. Therefore, the apoptosis-inducing dose of this quinolone, effective in antitumor therapies, equals at least 200–300 μg/mL.…”

“… 39 It has been estimated recently that the fluoroquinolone derivative induces oxidative stress in hepatoma-derived cells, increases activation of caspases involved in different pathways of cell death, and imposes double-strand breakages in the cellular DNA. 37 Moreover, CP triggers apoptosis of human triple-negative breast cancer (MDA-MB-231) cells by leading to a loss of the mitochondrial transmembrane potential and stimulation of the cell cycle arrest at the S-phase. 6 Interestingly, the effect of this antibiotic in prostate cancer (PC3) cells was mediated by the cell cycle arrest at the S-G2/M phase 41 and by inhibition of NF-κB binding to DNA.…”