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Published on: April 10, 2020
Screening of anti-cancer agent using zebrafish: comparison with the MTT assay
Yigen Li1, Wenjin Huang, Shenyuan Huang
1Drug Discovery Laboratory, School of Pharmacy, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, China.
Abstract:
The MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide) assay is a classical method for screening cytotoxic anti-cancer agents. Candidate drugs from the MTT assay need in vivo models to test their efficiency and to assess the absorption, distribution, metabolism, excretion, and toxicity of the drugs. An in vivo screening model could increase the rate of development of anti-cancer drugs. Here, we used zebrafish to screen a library of 502 natural compounds and compared the results with those from an MTT assay of the MCF7 breast cancer cell line. We identified 59 toxic compounds in the zebrafish screen, 21 of which were also identified by the MTT assay, and 28 of which were already known for their anti-cancer and apoptosis-inducing effects. These compounds induced apoptosis and activated the p53 pathway in zebrafish within 3h treatment. Our results indicate that zebrafish is a simple, reliable and highly efficient in vivo tool for cancer drug screening, and could complement the MTT assay.
Insights
Zebrafish offer a rapid in vivo method for anti-cancer drug screening, complementing traditional MTT assays. This study identified potent compounds, validating zebrafish as an efficient tool for cancer research.
Area of Science:
- Pharmacology and Toxicology
- Cancer Research
- Zebrafish Models
Background:
- The MTT assay is a standard in vitro method for initial screening of cytotoxic anti-cancer agents.
- Candidate drugs require in vivo validation for efficacy and pharmacokinetic profiling (ADME/Tox).
- Developing efficient in vivo screening models is crucial for accelerating anti-cancer drug development.
Purpose of the Study:
- To evaluate the zebrafish model as an in vivo screening tool for anti-cancer compounds.
- To compare the efficacy of zebrafish screening with the traditional MTT assay.
- To identify novel anti-cancer agents from a natural compound library using zebrafish.
Main Methods:
- A library of 502 natural compounds was screened using zebrafish as an in vivo model.
- Results were compared against an MTT assay performed on the MCF7 breast cancer cell line.
- Compound-induced apoptosis and p53 pathway activation were assessed in zebrafish.
Main Results:
- Zebrafish screening identified 59 toxic compounds, with 21 overlapping with MTT assay results.
- 28 identified compounds were previously known for anti-cancer and apoptosis-inducing properties.
- Tested compounds induced apoptosis and activated the p53 pathway in zebrafish within 3 hours.
Conclusions:
- Zebrafish serve as a simple, reliable, and highly efficient in vivo model for cancer drug screening.
- The zebrafish model effectively complements the MTT assay in identifying potential anti-cancer agents.
- This approach can accelerate the discovery and development pipeline for new cancer therapies.
