Nowadays, with genetic engineering methods, the production of transgenic medicinal plants has been provided to improve several traits such as increasing of valuable secondary metabolites, resistance to biotic and non-biotic stress. one of the most important applications of these techniques is to increase of the valuable pharmaceutical substance with manipulation in its synthetic pathway in plants. medicinal plants are the most important sources of anticancer compounds, natural compounds such as alkaloids, terpenes, lignans and flavonoids that are found in plants with high cytotoxicity in nature. one of the anticancer effects of these plants is the induction of apoptosis, which causes the death of cancer cells. trachyspermum ammi is a well-known medicinal plant in iran with many pharmaceutical properties. this plant contains a significant substance called thymol. thymol is an important phenolic compound used to treat many diseases, such as gastrointestinal disorders, lack of appetite, respiratory problems, asthma, and as an antiseptic for wound healing. many pharmaceutical studies were done on this plant and showed no toxicity for human utilization.
Methods
At first genetic construct of pbi121-tps was produced. then this construct was transferred to trachyspermum ammi ecotype. to confirm the increasing thymol contains and producing transgenic plants, pcr and hplc analysis were done. then alcoholic extract from leaves of transgenic and non-transgenic of trachyspermum ammi were acquired. the gastric cancer cell line (ags) was purchased from the iranian biological resource center (ibrc). several concentrations of extract solution were prepared (100 - 200 - 400 - 800 - 1000 , 2000 μg /ml). the cells were cultured in 24-well plate for 24 and 48 hours and incubated at 37 °c, 5% carbon dioxide. the cytotoxicity effects of the various concentration of thymol on ags cell were examined by mtt assay test and flow cytometry determined the apoptotic cell death rate
Results
Our finding demonstrated that the extract of transgenic trachyspermum ammi has a higher anticancer effect on ags cancer than the non-transgenic plant by suppressing cancer cell growth. extract of the transgenic plant with increasing concentration significantly decreased cell viability compared to control plant extract. also, with increasing incubation time, the cytotoxic effect of the extract enhanced and was a dose and time-dependent manner. the cancer cells exposed to the transgenic extract exhibited high apoptosis compared to non-transgenic extracts.
Conclusion
In this research, after confirmation of production of the transgenic plant, the anticancer effects of its extract on the gastric adenocarcinoma cancer were investigated. our results indicated that this extract has a high ability to produce apoptosis in cancer cell lines than non-transgenic plant extracts. due to the fact that the transgenic plant extract has high apoptosis effects on cancer cells, the cytotoxic compounds present in the extract of this plant can be a stimulant for the isolation, purification and identification of new and effective drugs in the treatment of gastric cancer and thymol might use as a tentative agent in the future to treat cancer.
