Scutellarin Inhibits The Proliferation And Bcl-2，Bax Expression Of Human Tongue Carcinoma Tca8113Cells

Objective:Tongue squamous carcinoma cell (TSCC) is one of the mostcommon malignant tumors in oral and maxillofacial region, have a higherrelapse rate and the rate of lymph node metastasis and poorprognosis,serious threat to human health. Scutellarin are flavonoidsextract from the stems and leaves of Scutellaria. Modern pharmacologicalstudies confirmed that the flavonoids in many ways have a certain role inthe anti-inflammatory, anti-allergic, anti-microbial and anti-tumor. Inrecent years, its anti-tumor activity got to the increasing emphasis. Tostudy the role of the scutellarin in TSCC, this experiment selects theTca8113cell lines to analysis the mechanisms that scutellarin inhibit cellproliferation and induce apoptosis of tongue squamous carcinoma cell.Methods:1.Tongue cancer cells Tca8113were bought in nanjing KaiJibiotechnology limited company, were cultured in vitro used forexperiments.2.Experimental group: divided into control group, differentconcentration scutellarin group. Control group was treated with10%FBSof RPMI-1640culture medium, while scutellarin groups with80μg/ml,120μg/ml,160μg/ml’s scutellarin.3.MTT colorimetric assay was applied to investigate the effects ofscutellarin on the proliferative activity of the Tca8113cells in12,24,48hours.4.Immunocytochemical staining: the expression of Bcl-2,Bax ineach group were detected by Immunocytochemical staining.5.Western blot: the expression of Bcl-2,Bax in each group were detected by Western blot.6.PT-PCR: the expression of Bcl-2,Bax in each group were detectedby PT-PCR.7.The statistical analysis was performed with SPSS17.0of statisticalsoftware. All the data was analyzed by Analysis of variance and LSD-ttest.P<0.05was considered statistically significant.Results:1.The impact of scutellarin on the value-added activity of Tca8113cells.Treated with80μg/ml,120μg/ml and160μg/ml concentration ofscutellarin for12h,24h and48h respectively, the growth of Tca8113cellshad showed time-dependent and dose-dependent manner. Throughstatistical analysis, the difference was statistically significant betweengroups (P <0.05).2.Immunocytochemical staining test showed that: under the action ofdifferent concentrations of scutellarin (80μg/ml,120μg/ml,160μg/ml),each group of Tca813cells cultured for48h, then detected byimmunocytochemical staining, and positive staining was mainly localizedin the cytoplasm. With the concentration of scutellarin increasing, Bcl-2positive staining becomes weaker and weaker,Bax positive stainingbecomes stronger and stronger. Through statistical analysis, the differencewas statistically significant between groups (P <0.05).3.Western blot test showed that: under the action of differentconcentrations of scutellarin (80μg/ml,120μg/ml,160μg/ml), each groupof Tca813cells cultured for48h,the relative expression of Bcl-2in eachgroup of Tca8113cells were1.172±0.026,1.074±0.069,0.776±0.041,0.566±0.054,the control group high expressed Bcl-2, and with theconcentration of scutellarin increasing, the relative expression of Bcl-2decreased. The relative expression of Bax in each group of Tca8113cellswere0.356±0.028,0.451±0.044,0.698±0.078,0.779±0.035,the controlgroup low expressed Bax, and with the concentration of scutellarin increasing, the relative expression of Bax increased, which showed adose-dependent tendency. Through statistical analysis, the difference wasstatistically significant between groups (P <0.05).4.RT-PCR test showed that: under the action of differentconcentrations of scutellarin (80μg/ml,120μg/ml,160μg/ml), each groupof Tca813cells cultured for48h, the Bcl-2mRNA expression in eachgroup of Tca8113cells were0.975±0.069,0.850±0.073,0.776±0.082,0.575±0.047, the control group high express Bcl-2, and with theconcentration of scutellarin increasing, the Bcl-2mRNA expressiondecreased. The BaxmRNA expression in each group of Tca8113cellswere0.769±0.061,0.817±0.121,0.895±0.044,0.975±0.115, the controlgroup low express Bax, and with the concentration of scutellarinincreasing, the Bax mRNA expression increased, which showed adose-dependent tendency. Through statistical analysis, the difference wasstatistically significant between groups (P <0.05).Conclusion:1. Scutellarin inhibited the growth of Tca8113cells and induced theapoptosis.2.Its mechanism may be related to downregulation of Bcl-2expression and upregulation of Bax expression.