Diallyl Disulfide에 의한 인체 자궁경부암 세포종 HeLa에 대한 세포사멸 기전연구

Abstract

Abstract

Introduction: It is reported that Diallyl Disulfide (DADS) known as one of the major components of garlic, has antimutagenic, anticoagulant, antibacterial and anticarcinogenic effects. Apoptosis is the process of programmed cell death that may occur in multicellular organisms and is important to sustain homeostasis and also known as one of the important mechanisms of treating cancer. Programmed cell death involves a series of biochemical events leading to a variety of morphological changes including cell shrinkage, nuclear fragmentation, chromatin condensation and chromosomal DNA fragmentation. In this study, I assessed cell death mechanism of HeLa which triggered by DADS administration. Methods: MTT assay was performed to evaluate cell viabilities, genomic DNA fragmentation assay to classify apoptosis from cell death, flow cytometry to assess cell death cycle and western blotting to evaluate apoptosis related proteins. Results: Cell viability was significantly decreased in DADS (100uM) treated HeLa cell group compared to CTL after 24 hours of incubation (p<0.01). Increase of DNA fragmentation was confirmed through electrophoresis in DADS (100uM) treated group, and also apoptoticbody formation and chromatin condensing were confirmed through fluorescence microscope. In western blotting analysis, pro-apototic agent Bax and caspase-3 and caspase-9 significantly increased compared to CTL respectely, While anti-apoptotic agent, Bcl-2 was significantly decreased compared to CTL . PARP segregation was also observed. Conclusion: This study suggests that DADS induced cell death of HeLa is processed through apoptotic pathway and DADS may be a potent anticarcinogen for human cervical cancer.

Key word: HeLa cells, Diallyl Disulfide(DADS), Bax, Bcl-2, caspase-3, caspase-9, PARP