The role of in-vitro MTT experiment in preclinical efficacy evaluation of anticancer drugs

Practical methods for evaluating tumor cell growth and survival are very important in tumor research.In preclinical efficacy evaluation of antitumor drugs, in vitro MTT pharmacodynamics experiments are often used to explore the inhibitory effect of compounds on the growth of tumor cells in vitro, or in vivo tumor transplantation pharmacodynamics experiments are used to explore the inhibitory effect of compounds on the growth of tumor transplanted animals. MTT colorimetry is a method used to evaluate the survival status of tumor cells, which has attracted much attention in recent years.

Tetramethylazolazole-blue (MTT) colorimetric method can reflect the changes of tumor cells by transforming MTT into its formazan deoxygenase activity when the appearance of tumor cells has not changed significantly under light microscope. It has been widely used in the study of anti-tumor drugs.MTT assay was used to observe the inhibitory effect of the compounds on the growth of tumor cell lines in vitro, and to evaluate the antitumor activity of the compounds.

In preclinical pharmacodynamics evaluation, MTT colorimetry can be used to evaluate the cytotoxicity of compounds and the sensitivity test of antitumor drugs.If there are researchers in the in vitro study, taking the active fraction of andrographolia (AEP) as the model drug, taking mouse breast cancer cells 4T-1 and mouse lung cancer cells LLC as the model, using MTT assay to investigate the inhibitory effect of drugs on tumor cell proliferation.It was found that AEP inhibited the proliferation of tumor cells in a time-dependent and dose-dependent manner, and the difference was significant compared with blank control group.Medicil provides drug safety evaluation services. It has established a drug safety evaluation system that meets international standards, and its experimental studies follow the guidelines of ICH, CFDA, FDA and OECD. Medicil can provide customers with high-quality data, rapid study cycle and preclinical drug safety evaluation services.

Some researchers have also explored the proliferation of Dandelion Root Extract (DRE) and Viola odorifera Aqueous Extract (VE) on human breast cancer cell lines (MCF-7) and human liver cancer cell lines (HepG2) Inhibition rate and cell viability [1]. The researchers prepared DRE and VE with different drug mass concentration gradients, which acted on MCF-7 cells and HepG2 cells, and used cell morphology analysis, AO/PI double staining method, MTT method, and mitochondrial membrane potential detection method to explore the two types of antibodies. The effects of cancer medicinal plants on the growth viability, inhibition of proliferation and changes in mitochondrial membrane potential of two kinds of cancer cells. Results The MTT method showed that after 48 hours of drug action, DRE had the better inhibitory effect on MCF-7 cells. The inhibitory effect was the best when the mass concentration was 6.0 mgmL-1, and the cell lethality rate reached 92.53%; for HepG2 The cell inhibitory effect is better for VE. The best inhibitory effect is when the mass concentration is 6.0 mg•mL-1. The cell lethality rate is 93.46%, and the cell viability of the two types of cells decreased, and the mitochondrial membrane potential also showed a downward trend.

Some researchers also extracted the active components from Bupleurum spp. callus (BSW-ws) and administered SMMC-7221 to cultured liver cancer cells at concentrations of 10, 20, 30, 40, 60, 80, 100 μg•mL-1 ; MTT method was used to detect the cytotoxic activity of BSW-ws, and the effect of BSW-ws on cell apoptosis was detected by flow cytometry [2]. The results of detecting the cytotoxic activity of BSW-ws by MTT method showed that the IC50 value was reached when the administration concentration was 60μg•mL-1.

MTT colorimetry is a rapid and quantitative method to detect the number of living cells, and its results are relatively reliable. It has been widely used in cell experiments.MTT assay can be used for tumor and fungal cell activity detection, drug sensitivity detection of tumor cells, large-scale screening of anti-tumor, etc. Therefore, in vitro MTT pharmacodynamics experiment is also widely used in preclinical pharmacodynamics evaluation.It is believed that with the development of science and technology and the continuous improvement of MTT colorimetry, its application in the field of drug research and development will be more and more extensive, and the role will be more and more significant.

[1] Study on the anticancer activity of the water extracts of Dandelion and Viola serrata [J].

[2] Study on the anti-hepatocarcinoma effect of Bupleurum callus extract[J].