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A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules
Published on: March 10, 2017
Benzethonium chloride: a novel anticancer agent identified by using a cell-based small-molecule screen
Kenneth W Yip1, Xinliang Mao, P Y Billie Au
1Department of Medical Biophysics, University of Toronto, and Department of Medical Oncology, Princess Margaret Hospital, University Health Network, Toronto, Ontario, Canada.
Purpose:
This study aims to identify a novel therapeutic agent for head and neck cancer and to evaluate its antitumor efficacy.
Experimental Design:
A cell-based and phenotype-driven high-throughput screening of approximately 2,400 biologically active or clinically used compounds was done using a tetrazolium-based assay on FaDu (hypopharyngeal squamous cancer) and NIH 3T3 (untransformed mouse embryonic fibroblast) cells, with secondary screening done on C666-1 (nasopharyngeal cancer) and GM05757 (primary normal human fibroblast) lines. The "hit" compound was assayed for efficacy in combination with standard therapeutics on a panel of human cancer cell lines. Furthermore, its mode of action (using transmission electron microscopy and flow cytometry) and its in vivo efficacy (using xenograft models) were evaluated.
Results:
Benzethonium chloride was identified as a novel cancer-specific compound. For benzethonium (48-hour incubation), the dose required to reduce cell viability by 50% was 3.8 micromol/L in FaDu, 42.2 micromol/L in NIH 3T3, 5.3 micromol/L in C666-1, and 17.0 micromol/L in GM05757. In vitro, this compound did not interfere with the effects of cisplatin, 5-fluorouracil, or gamma-irradiation. Benzethonium chloride induced apoptosis and activated caspases after 12 hours. Loss of mitochondrial membrane potential (DeltaPsiM) preceded cytosolic Ca2+ increase and cell death. In vivo, benzethonium chloride ablated the tumor-forming ability of FaDu cells, delayed the growth of xenograft tumors, and combined additively with local tumor radiation therapy. Evaluation of benzethonium chloride on the National Cancer Institute/NIH Developmental Therapeutics Program 60 human cancer cell lines revealed broad-range antitumor activity.
Conclusions:
This high-throughput screening identified a novel antimicrobial compound with significant broad-spectrum anticancer activity.
Insights
Benzethonium chloride, an antimicrobial agent, shows significant anticancer activity against head and neck cancers. This compound effectively reduces tumor growth and induces apoptosis, demonstrating broad-spectrum efficacy in preclinical models.
Area of Science:
- Oncology
- Pharmacology
- Biochemistry
Background:
- Head and neck cancers represent a significant global health challenge.
- Novel therapeutic agents are urgently needed to improve treatment outcomes.
Purpose of the Study:
- To identify a novel therapeutic agent for head and neck cancer.
- To evaluate the antitumor efficacy of identified compounds.
Main Methods:
- High-throughput screening of approximately 2,400 compounds using cell-based assays.
- Secondary screening on cancer and normal cell lines.
- In vitro and in vivo efficacy studies, including combination therapy and mode of action analysis.
Main Results:
- Benzethonium chloride identified as a novel, cancer-specific compound.
- Demonstrated dose-dependent cytotoxicity in head and neck cancer cell lines.
- Induced apoptosis, activated caspases, and caused loss of mitochondrial membrane potential.
- Showed in vivo efficacy in xenograft models and broad-spectrum activity against NCI-60 cancer cell lines.
Conclusions:
- Benzethonium chloride exhibits significant broad-spectrum anticancer activity.
- This compound represents a promising novel therapeutic candidate for various cancers.
