Apoptosis induction of the essential oil from the leaf of pycnocycla bashagardiana mozaff. in ht-29 cells: association with expression bcl-2 and bax

Shima Aramideh ,1 Masoumeh heshmati,2,* Jinous asgarpanah,3

1. pharmacy student, Pharmaceutical Sciences Research Center, Tehran medical Sciences, Islamic Azad university,Tehran ,Iran
2. Department of Molecular and Cellular Sciences, Faculty of Advanced Science & Technology ,Tehran medical Sciences, Islamic Azad university,Tehran ,Iran
3. Department of Pharmacognosy, Faculty of Pharmacy,Tehran medical Sciences, Islamic Azad university,Tehran ,Iran

Abstract

Introduction

Pycnocycla bashagardiana mozaff is a rare endemic species that has been used in folk medicine in south of iran. it is commonly distributed in jask county, hormozgan province. due to the presence of myristicin as the major component (more than 65 % of oils)cytotoxicity and anti-cancer properties of p. bashagardiana essential oil are proposed.on the other hand colorectal cancer is the third most lethal cancer in men and fourth in women. so far, various drugs have been used to treat the disease, but because of the emergence of drug resistance to chemical drugs, the need to achieve the new natural compounds of medicinal plant derivatives is felt more active. due to the presence of other substances, the herbal substance has a balanced biological state, so it does not accumulate in the body and has no side effects and can play an important role in the treatment of cancer. the aim of this study was to investigate the effect of pycnocycla bashagardianaʼleafs cytotoxicity and anti-cancer on ht29 cells and l929 mouse fibroblast cells in vitro

Methods

In this study, the cytotoxicity effects of pycnocycla bashagardianaʼleafs was studied at different concentrations 3.1-500 µg/ml of essential oils on ht29 and normal l929 cells at 48 h by trypan blue exclusion assay and mtt assay .apoptosis induction was determined by flow cytometric analysis (annexin-v-fitc). the expression levels of bax and bcl2 in ht29 cells and normal l929 cells were detected using quantitative real- time pcr (qrt-pcr)

Results

Pycnocycla bashagardianaʼleafs has cytotoxic effects depending on concentration and time. ic50 values were observed at concentration of 52 μg / ml at 24h and 12.5 μg / ml at 48,72 h in the ht29 cells and at 500 and 50 μg / ml at 48 and 72h and no cytotoxicity was recorded at 24 h in l929 cells by trypanblu exclusion assay. according result by mtt assay, ic50 values were recorded at 170, 72 and 12.5 at 24,48 and 72h in ht29 cells and 500, 100 and 50 at 24,48 and 72 in l929 cells. apoptosis results were resulted 1.2 , 2.27 and 2.06 fold in ht29 cells and 1.3, 1.45 and 1.62 in l929 cells at concentration 6.25, 12.5 and 25 μg / ml. the decreased and increased expression level of bax and bcl2 was recorded in ht29 cells and l929 cells , as whole the ratio of bax?bcl2 in ht29 cells was more than l929 cells.

Conclusion

Comparison results by trypan blu exclusion, mtt, apoptosis and gene expression assays suggests that the pycnocycla bashagardianaʼleafs has toxicity in cancerous cells and normal cells,but can induced apoptotic cell death in cancerous cell more than normal cells.

Keywords

Apoptosis, pycnocycla bashagardiana mozaff, bcl-2 -bax