The effects of anticancer of 7-hydroxycoumarine on the colon cancer cell line (ht29) and the normal mouse fibroblast cells (l929)

Fatemeh Karami,1,* Masoumeh heshmati ,2 Shima tavakol,3 Parvaneh najafizadeh,4

1. MSc, Pharmaceutical Sciences Research Center, Pharmaceutical Sciences Branch, Islamic Azad university, Tehran-Iran
2. Assistant Professor, Department of Molecular and Cellular Sciences, Islamic Azad University, Tehran-Iran
3. Assistant Professor, Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran-Iran
4. Assistant Professor, Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran-Iran

Abstract

Introduction

colorectal cancer it is a lethal and common cancer. due to the development of drug resistance to chemical drugs, the need to achieve new natural compounds of medicinal plants derivatives is felt more active. herbal substance due to being combined with other ingredients, is consistent of a biological equilibrium, so it does not accumulate in the body and has no side effects and can play an important role in the treatment of cancer. coumarin is a mixture of the family of phenolic compounds in plants. these phenolic substances are physiologically active and have toxic effects on the living cells that interfere with the nuclear division, growth and inhibition of enzymes such as amylases or sucralose (enuresis). the aim of this study was to investigate the effect of the anticancer of a coumarin derivative called 7-hydroxycoumarin(7-hc) on colon cancer cells ht29 and normal mouse fibroblast cells l929 in vitro.

Methods

In this study, the effects of 7-hc cytotoxicity at different concentrations on proliferation and survival of ht29 and l929 cells were monitored with mtt assay and trypan blue the concentration of active ingredient which causes the death of half the cells (ic50). the number of 5 × 104 cell was planted from 24 houses and then the values of 12.5-250 μg/ml of the effective 7-hc after one day from the date of cell culture. subsequently, the active ingredient was placed adjacent to the cells for 24, 48 and 72 h. then, the number of live cells and the mitochondrial dehydrogenase activity of these cells were evaluated using trypan blue exclusion assay, mtt assay. flow cytometry was used to evaluate the cell cycle after treatment with 7-hc. flow cytometry and pi staining was performed in order to evaluate the cell cycle of ht29 and l929 cell line after treatment with 7-hc. cell cycle analysis was accomplished with using propidium iodide. 1.5× 106 cells/ml were treated(50, 100 and 250μg/ml) and without 7-hc concentrationat at 48h.

Results

To help using the methods in this study, according to the results of the data article 7-hc has cytotoxic effects depending on concentration and time. ic50 values were observed at a concentration of 100 μg/ml in the ht29 and 250μg/ml in the normal l929 cell line(p<0.001). respectively, the analyses of cell cycle in the ht29 and l929 by flow cytometry showed arresting of cells in g0g1 phase at 100 and 50 μg/ml of 7-hc on ht29 (p<0.01), but no effect on normal cells was observed.

Conclusion

Comparison of ic50 values suggests that the 7-hc has a low concentration of specific toxicity on cancer cells and potentials of cellular toxicity and anti-cancer effects on colon cancer cells. also, this active ingredient only stopped the cell cycle of the cancer cells which is very important in the treatment of cancer.

Keywords

Colon cancer, ht29 cell line, l929 cell line, coumarin