| Objective:To detect the apoptosis and cell growth inhibition in vitro effects of Artesunate(ART)on novel human palatal salivary gland cell line of adenoid cystic carcinoma(NACC),and to evaluat its antitumor effect of artesunate on nude mice bearing NACC cells transplantation tumor model,and to explore the possibility and related mechanism of artesunate as adenoid cystic cancer treatments.Method:Cell growth inhibition ratio were determined by the methyl thiazolyl tetrazolium(MTT)method in vitro after NACC cell line were treated with various concentrations of ART.Clone formation experiment was used to detect the effect of cell proliferation.The morphological changes of NACC cell line was observed by light microscopy and electronmicroscop.The effect of ART on cell migration ability of NACC cells were detected by wound healing method.Flow cytometry(FCM)assay was used to analyze the cell cycles and cell apoptosis. NACC cancer mouse model with were randomly divided into six groups as the control group,the ART50.0,100.0,200.0mg/kg groups,the DDP2.0mg/kg group and the ART50.0mg/kg+DDP1.0mg/kg group.Treatment start at the7th day when solid tumors were palpated.Drugs were given once every2day for16days,The size of the subcutaneous tumor were measured,inhibitory rate was calculated.Animals were sacrified and tumor were harvest weighted on the23th day after cancer cell innoculation.Tumor cell morphology was observed through HE staining.Immunohistochemistry was used to determined the expression of Ki-67and CXCR4in tumors of each group.Result:Experiment of NACC cell in vitro:(1)The proliferation of NACC cell line was suppressed by ART.IC50of artesunate in24,48and72h were113.09,74.80,35.61μg/ml respectively.And this inhibitory action was positive correlated with time-and dose-dependent CPO.05).ART could inhibit the proliferation of NACC cells.(2)There were typical apoptosis structure could be observed in ART groups such as organelles reduced,chromatin pyknosis and the formation of apoptotic bodies.(3)Meanwhile,ART significantly inhibited the cell migration.(4)No significant difference had been detect between the ART12.5u g/ml treated group and the control group in the apoptosis ratio.With the increase of concentration of ART,apoptosis ratio of NACC cell was also elevated.The ART25.0μ g/ml group,ART50.0Ug/ml group,ART100.0μg/ml group,ART200.0μ g/ml group might induce apopotosis significantly at a ratio of(17.92±0.26)%,(27.95±0.40)%,(58.91±3.81)%,(89.58±2.94)%respectively.(5)ART could arrest the cell cycle in G)phase at the dosage groups of12.5,25.0and50.0μg/ml and in G2phase at50.0,100.0and200.0μg/ml groups. Results of transplantation tumor model of NACC cell:(1)NACC cancer mouse model with solid tumor were constructed,the tumor inhibition rates of ART50.0,100.0,200.0mg/kg groups,the DDP2.0mg/kg group and the ART50.0mg/kg+DDP1.0mg/kg group were36.01%,48.95%,78.67%,64.69%and63.29%.(2)Tumor cells grew vigorously in the control group,lots of apoptosis and necrosis cells were observed in the other groups,especially in ART200.0mg/kg group.The expression of Ki-67in ART200.0mg/kg group,DDP group and conbination group were decreased significantly compared with control group,and the expression of CXCR4in ART200.0mg/kg group,ART100.0mg/kg group,DDP group and conbination group were decreased significantly compared with control group.Conclusion:(1)ART can inhibit the growth and induce apoptosis of human palatal salivary gland NACC cell line.With the increase of concentration of ART,apoptosis ratio of NACC cell was also elevated,this action was positive correlated with dose-dependent(P<0.05)(2)ART could arrested the cell cycle in G1phase and G2phase of NACC cell.(3)ART can inhibit the growth of nude mice xenograft of NACC cell,increase the sensitivity of NACC cell to cisplatin,downregulation the expression of Ki-67and CXCR4in transplanted tumor. |
