The Effects Of Scutellarin On The Expression Of Survivin And Caspase-8in Human Tongue Squamous Carcinoma Tca8113Cell

Tongue cancer is the most common oral cavity cancer. The incidenceof the tongue squamanous cancer accounts for39.95%in oral cancerdisease in our country. It is the first incident in oral cavity cancer disease.Most tongue cancer is squamanous carcinoma. It is characterized bygrowing quickly, strong invasion, high malignant degree, etc. The lymphnode in tongue cancer patient has the high metastasis rate. In theadvanced stage, cancer cells will transfer to the lungs or other parts of thebody. At present, the management of tongue cancer is to treat by surgeryprimarily, and complementary with radiation and chemotherapy. However,the recurrence and metastasis rate after surgery are high. Therefore, it hasthe positive significance to seeking the highly effective and low adversereaction medicine to treat tongue cancer for improve the patients’ lifequality and enhancing the survival rate.The scutellarin, a flavoniod, extracted from stem-leaf of Scutallariabaicalonsis George, has many actions such as antibacterial action, diureticaction, anti-inflammatory action, anti-immune action and anticonvulsion,ect. And it also has many physical effects such as: strong anti-oncologyaction. This study discusses scutellarin to induce the mechanism ofhuman tongue squamanous cancer apoptosis by observing apoptosis andthe change of survivin and caspase-8expression using scutellarin to affecthuman tongue squamanous cancer Tca8113cultured in vitro by. Thisstudy provides the theoretical foundation and experimental data for theanti-oncology action of scutellarin.Objective:To investigate the effects and mechanism of scutellarin on apoptosisand the change of survivin and caspase-8expression of Human tongue squamous cell carcinoma Tca-8113cell.Methods:1. Tongue cancer cells Tca8113were bought in nanjing KaiJibiotechnology limited company, used for training, experimental need.2. Experimental group: divided into normal control group, differentconcentration scutellarin group. The control group was treated with10%FBS of1640culture medium, while scutellarin groups with80μg/ml,120μg/ml,160μg/ml’s artemether cultures of cells according to the resultof preliminary experiment.3. Tunel assay was applied to investigate the effects of scutellarin onthe early apoptosis of the Tca8113cells.4. Immunocytochemistry method was applied to observe the effectof scutellarin on survivin and caspase-8expression in Tca8113cells.5Western-blot method was applied to examine the expression ofsurvivin and caspase-8in Tca8113cells6. The statistical analysis was performed with SPSS17.0usingone-way analysis of variance (ANOVA). P <0.05was consideredstatistically significant.Result:1Tunel analysis showed that: the control group and80μg/mL,120μg/mL,160μg/mL of the scutellarin Tca8113cells treated with for48h,Control group can be seen scattered in the nucleus was dark brown orbrown apoptotic cells, the apoptotic index is4.45±0.27%. With theconcentration of scutellarin increasing, the apoptotic index wassignificantly increased. The apoptotic index were17.63±0.25%、36.23±0.36%、51.84±0.58%.Through statistical analysis, the difference wasstatistically significant between groups (P <0.05).（Fig.1～5和Table.1）2Immunocytochemistry analysis showed that: the control group and80μg/ml,120μg/ml,160μg/ml scutellarin treated with Tca8113cellsfor48h, then detected by immunocytochemical staining, under themicroscope positive staining was mainly localized in the cytoplasm. With the concentration of scutellarin increasing, survivin positive stainingbecomes weaker and weaker, caspase-8positive staining becomesstronger and stronger. Through statistical analysis, the difference wasstatistically significant between groups (P <0.05).（Fig.6～13和Table.2~3）3Western-Blot analysis showed that: the control group and80μg/mL、120μg/mL、160μg/mL of the scutellarin Tca8113cells treatedwith for48h, the control group high expressed survivin, and with theconcentration of scutellarin increasing, the relative expression of survivindecreased. The control group low expressed caspase-8, and with theconcentration of scutellarin increasing, the relative expression ofcaspase-8increased. Through statistical analysis, the difference betweenthe groups were statistically significant (P <0.05).（Fig.14a~b、Fig.15a~b和Table.4~5）Conclusion:Scutellarin inhibited the growth of Tca8113cells and induced theapoptosis, and its mechanism may be related to downregulation ofsurvivin protein expression and upregulation of caspase-8expression.