Tca8113Its Mechanism Of Apoptosis Induced By EGCG Oral Squamous Cell Carcinoma Cells

Objective: To investigate the green tea extract epigallocatechin table gallate ((-)epigallocathechin-3-gallate, EGCG) Tca8113human oral squamous cell carcinomacell proliferation, apoptosis as well as caspase-3, caspase-8protein expression ofapoptosis affected. In order to improve the theoretical basis for the EGCG in theprevention and treatment of oral squamous cell carcinoma.Methods:Different doses of EGCG (0,25,50,75,100,125mg/L),respectively, therole Tca8113cells24h,48h,72h, MTT method to observe the effect of the drug onTca8113cell growth, cell proliferation was calculated inhibition.Observe EGCG(0,50,75,100mg/L) for48h Tca8113morphological changes of cells under aninverted microscope.AO/EB fluorescence staining of different doses of EGCG(0,25,50,75,100mg/L), respectively, the role of the morphological changes of thecells after48h Tca8113cells under a fluorescence microscope.Hochest33258stainingdifferent doses of EGCG (0,25,50,75,100,125mg/L) on apoptosis Tca8113’s.Flowcytometry EGCG (0,50,75mg/L) after the intervention48h Tca8113cell cycle.TEMobservation of the difference between dosing groups with and without the drug group,which dosing group whether apoptosis.SABC immunocytochemistry assay withdifferent concentrations of EGCG (0,25,50,75,100,125mg/L) after the intervention48h, caspase-3, caspase-8protein expression differences in Tca8113cells. Results:MTT assay showed that different doses of EGCG (25,50,75,100,125mg/L)inhibited proliferation Tca8113oral squamous cell carcinoma cells, which(50,75,100,125mg/L) with the control group, the difference was statisticallysignificant (p<0.05), cell proliferation inhibition rate increased with increasingconcentrations of EGCG and time while increasing, showing a significant period oftime-dose-response relationship.inverted microscope Tca8113seen in the controlgroup were spindle or polygonal cells adherent growth, intensive quantity, outlineclear, intercellular connections are tight. Dosing group Tca8113cells with increasingconcentrations of EGCG, the number of adherent cells was significantly reduced,refraction poor, some cells became round, shrunken and floating in the medium.AO/EB staining showed different concentrations of EGCG intervention Tca8113cellsappeared typical apoptotic morphological changes compared with the control group,with increasing concentrations of EGCG, increasing the number of apoptoticcells.Hochest33258staining showed different concentrations of EGCG interventionTca8113cells appeared typical apoptotic morphological changes such as nuclearcondensation, nuclear fragmentation. Compared with the control group, withincreasing concentrations of EGCG, increasing the number of apoptotic cells (n=5, p<0.01).Using different concentrations of EGCG (0,50,75mg/L) for48h aftertreatment Tca8113for cell cycle analysis by flow cytometry, the percentage of cellsshowed that EGCG (50,75mg/L) was treated group G1of41.997±0.17and65.24±0.632, and the control group (not dosing group) comparing with significantdifferences (p <0.01), showed that EGCG induced G1arrest Tca8113cells.UnderTEM found that compared with the control group, dosing group (75mg/L) showedtypical apoptotic changes.Cells showed immunochemical test results, EGCG canincrease Tca8113cells caspase-3, caspase-8protein expression in a dose-dependentmanner. Different concentrations of EGCG (0,,25,50,75,100mg/L) interventionTca8113cells48h, the average optical density caspase-8protein (AOD) values were:0.131±0.04,0.177±0.019,0.223±0.06,0.376±0.074,0.571±0.085, each dosinggroup compared with the control group difference was statistically significant (p<0.01), the mean optical density of caspase-3protein (AOD) values were:0.096± 0.072,0.118±0.049,0.197±0.081,0.343±0.033,0.653±0.065, each dosing groupcompared with the control group difference was statistically significant (p <0.05).Conclusion: EGCG can effectively inhibit the proliferation of Tca8113cells,induce apoptosis, which may increase caspase-8, caspase-3expression.