The Influence To The Growth Of Squamous Cell Carcinoma Of Tougue (Tca8113)under Brazilin Combined With Pingyangmycin

Objective:To observe affection of the pingyangmycin and Brazilin proliferation and apoptosis of Tca8113cells in vitro, as well as combinate them, provides a new method with clinical combination chemotherapy for oral squamous cell carcinoma (SCC).Methods:To detect pingyangmycin and Brazilin by MTT method influence of proliferation ability of Tca8113cells respectively; Flow cytometry detection the rate of apoptosis from pingyangmycin, Brazilin, and their combination; Real-time fluorescent quantitative PCR detection relative expression levels of mRNA of apoptosis gene caspase3and caspase-9.Results:1. Brazilin, play a role of inhibition the proliferation of Tca8113, under the same conditions with same concenteation of braziln, the inhibition rates of Tca8113cells become greater with times go on; under the same conditions with same time, the greater the inhibition rate of Tca8113cells with the increase of concentration; the stronger the inhibition. According to the concentration inhibition rate curve, at72h, IC50,146ug/ml Brazilin grain the best concentration (150ug/ml).2. Pingyangmycin, play a role of inhibition the proliferation of Tca8113, under the same conditions with same concenteation of Pingyangmycin, the inhibition rates of Tca8113cells become greater with times go on; under the same conditions with same time, the greater the inhibition rate of Tca8113cells with the increase of concentration; the stronger the inhibition. According to the concentration-inhibition rate curve, at72h,IC50,16ug/ml Pingyangmycin grain the best concentration (10ug/ml).3. The results of Flow cytometry showed the percentage of apoptosis respectively after use the medication72hours, blank control group (A), Brazilin group(B), pingyangmycin group(C) and the combination group(D):1.25±0.21,11.36±0.46,14.27±0.51and22.17±0.67, group of B,C, D Tca8113cells have obvious trend of apoptosis, including group D Tca8113cells apoptosis percentage increased significantly, higher than in B and C group (p<0.01).4. The results of RT-PCR showed that, after use the medication72hours, the relative expression of caspase-3mRNA in group B, C, D is greater than blank control group (A)(p<0.05), and group D is significantly higher than group B and C (p<0.05), while B and C group had no significant difference (p>0.05); the relative expression of caspase9mRNA respectively is:group B, C, D is significantly higher than group A (p<0.05), group D is significantly higher than group B and C (p <0.05), while B and C group had no significant difference (p>0.05).Conclusion:1. Brazilin, play a role of inhibition the proliferation of Tca8113. At the same time, the higher the concentration, the stronger the inhibition; At the same concentration, the longer time, the stronger the inhibition.2. Pingyangmycin, play a role of inhibition the proliferation of Tca8113. At the same time, the higher the concentration, the stronger the inhibition; At the same concentration, the longer time, the stronger the inhibition.3. Comnbination therapy of Brazilin and pingyangmycin have a stronger effect of apoptosis of Tca8113cells.