Study of anti-cancer effects of Grandivittin on the apoptosis pathway in A549 cell line
Study of anti-cancer effects of Grandivittin on the apoptosis pathway in A549 cell line
Fateme Zomorodi Anbaji,1,*Seyed Jalal Zargar,2
1. Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran 2. Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran
Introduction: Homeostasis is maintained in normal tissues by a highly regulated process of cell proliferation. If there is an imbalance in the cell proliferation stage and the cell death stage, cancer develops. Lung cancer is the second most common cancer in men and women and the leading cause of death among cancers. The A549 cell line belongs to non-small cell lung cancer, accounting for 85% of all lung cancers.
Grandivittin is one of the Dihydrofuran coumarins. Dihydrofuran Coumarins are a group of Coumarins that are very active as anti-cancer and cancer preventers. It also has various effects, such as antioxidant and laxative effects. This study investigated the effects of different concentrations of Grandivittin on cell survival and death in the A549 lung cancer cell line. The results of this study show the impact of Grandivittine on the inhibition of A549 cancer cells. This study found that in the A549 cell line, the rate of cell death increased at 24 and 48 hours due to increasing concentrations of 1.3, 1, 0.7, 0.4, and 0.1 GRD. By performing MTT tests at a concentration of 0.7 in 48 hours, we reached 50% lethality and confirmed this result by flow cytometry test. We performed a real-time RT-PCR test to evaluate the function of Grandivitin on the expression of genes affecting apoptosis. During these experiments, we observed that BAX, TP53, CASP9, and CASP3 genes had a significant increase in mRNA level, and the BCL2 gene also had a decrease in mRNA expression. Based on the results obtained in this study, Grandivittin can be a promising substance for the treatment of cancer that needs further research.
Methods: Cell line A549 was treated under the effects of incremental concentrations of 1.3, 1, 0.7, 0.4 and 0.1 µl Grandivitin at times 24 and 48 hours after culture and passage. Then RNA was extracted, and cDNA was made. By performing MTT tests at a concentration of 0.7 in 48 hours, we reached 50% lethality and confirmed this result by flow cytometry test. We performed a real-time RT-PCR test to evaluate the function of Grandivittin on the expression of genes affecting apoptosis.
Results: In this study, we observed that at a concentration of 50% lethality, 0.7 µl, in 48 hours, BAX, TP53, CASP9, and CASP3 genes had a significant increase in mRNA level, and the BCL2 gene also decreased mRNA expression.
Conclusion: Our study shows that grandivitine inhibited the mRNA expression of BCL2 anti-apoptotic gene and increased the expression of BAX, TP53, CASP9 and CASP3 genes. Our results from this experiment are convergent with other studies on coumarin derivatives. Finally, it should be noted that grandivitine can be an effective substance in the treatment of cancer.