The Induction Of Apoptosis In Human Conlon Cancer Cell Line LS-174-T Through Upregulation Of Bax And Downregulation Bcl-2Gene By Allicin

According to its nature Cell death origin and biological significance divided into twokinds of completely different types：Apoptosis and necrosis.Apoptosis has an importantsignificance in tumors and other diseases. Currently, It carried out extensive research inmany areas of biology. It has made significant progress in the biochemical reactionmechanism of apoptosis and apoptosis gene regulation.AIM: The experiment is to study the antitumor effet to investigate the the Allicin byregulating bax/bcl-2gene expression induced apoptosis.METHODS: Observe the apoptotic morphology of the LS-174-T cells by invertedmicroscope, fluorescence microscopy and transmission electron microscopy. Agarose gelelectrophoresis technique for observation and analysis apoptosis and biochemicalcharacteristics.The flow cytometry (FCM) is used to investigate the changes of mitochondrialmembrene potential(DYm), the apoptotic rate and changes of cell cycle distribution.Bax/bcl-2mRNA was assayed by real-time fluorescent quantitative PCR. Western blotanalysis Bax/bcl-2protein expression levels.RESULTS:1.Allicin can inhibit and induce apoptosis in human colon cancer LS-174-T,The allicin best effect is18ug/mL after48hours.2.The IC50of the LS-174-T cells treatedwith Allicin for48h is20.292μg/mL.3. Flow cytometry results showed that allicinLS-174-T-cell cycle arrest in S phase and G2phase. Flow cytometry the Annexin VFITC/PIdouble labeled LS-174-T-cell apoptosis. With the drug concentration increase in thepercentage of apoptosis in early first increase and cytometry the Annexin VFITC/PI doublelabeled LS-174-T-cell apoptosis. With the drug concentration increase in the percentage ofapoptosis in early first increase and then reduce. The later Apoptosis percentage wasincreasing. The28μg/mL the role of allicin LS-174-T cells after48h of mitochondrialmembrane potential is110.36±0.96. Significantly reduced compared with the control group(p <0.05).4. Within a certain range bax mRNA expression increased with the increase of drugconcentration and showed a increasing trend. bcl-2mRNA expression with increasing drugconcentration and showed the weakening trend.5. Western blot showed that Bax proteinexpression gradually increased with the increase of drug concentrations of allicin. Bcl-2protein expression was gradually decreased. CONCLUSION: Allicin can induce apoptosis by up-regulating bax genes anddownregulation of bcl-2gene.