Reserch Of Proliferation Inhibiting And Apoptosis Inducting Effects Of Parthenolide On Adenoid Cystic Carcinoma Of Salivary Gland In Vitro

ObjectiveTo investigate the inhibition of proliferation and the induction of apoptosis ofParthenolide（Par） on Adenoid cystic carcinoma of salivary gland （SACC-83）cells in vitro. We clarified the effect of NF-кB、Livin and Caspase-3,and thepossible mechanism in Par induced SACC-83cell apoptosis.Methods（1）To observe the cell morphologic change after24h by Par（5.0mg/L）induced under invert microscope;（2）MTT was used to detect optical density（OD）of SACC-83cells by intervention of Par in six different concentration group in24,48and72h,The effect of Par on SACC-83cells proliferation was measuredusing MTT assay;（3）Propidium iodide（PI） single-staining flow cytometryassay method was used to analyze cell cycles of SACC-83cells by interventionof Par in three different concentration（7.5、15.0、30.0mg/L）groups. Toobserve cell percentage distribution in G0/G₁、 S and G₂/M phases byintervention of different Par concentration groups after24h;（4） the cell cycledistribution and apoptosis status were analyzed by Flow cytometry.（5） theexpression of NF-кB、Livin and Caspase-3protein were determined by westernblot. The data analyzed by SPSS13.0software package.Results（1）The morphological analysis demonstrated that the cell became round andsmall in shape,the quantity was reduced significantly,their nuclei exhibited afragmented;（2）MTT assay showed that Par inhibited SACC-83cells proliferationin time-and concentration-dependent manner. The inhibiting concentration of 50%cell growth（IC50）at24、48and72h was （60.23±7.41）、（27.44±5.45）and（8.73±5.02）mg/L;（3）As compared with normal control group in the samephase, cell percentage of G0/G₁phase in SACC-83cells were increasedsignificantly in three different Par concentration group（7.5、15.0、30.0mg/L）after administration for24h,with statistical significance（P<0.05）, at the sametime,cell percentage of S phase was reduced, cell percentage of G₂/M phasewas not changed. Thus,a concentration-dependent manner made SACC-83cells cycle arrest in G0/G₁phase.（4）Par could induced apoptosis of SACC-83cells,The cell apoptosis rates were increased gradually in three differentconcentration groups.the cell apoptosis rates were14.7%±2.1%、30.5%±3.4%and38.7%±2.7%respectively, all significantly higher than that of the controlgroup3.2%±0.9%,all P<0.05.（5）Western blot showed that Par could reducedthe expression of NF-кB、 Livin protein,increased Caspase-3protein in aconcentration-dependent manner（P<0.05）.Conclusionsparthenolide can inhibit the proliferation of SACC-83cells in vitro.and a largenumer of cells accumulate in G0/G₁phase, the apoptosis mechanism may berelated with down-regulate the expression of NF-кB、Livin protein and activationCaspase-3protein.