三氟拉嗪通过诱导p16^INK4a抑制BGC-823细胞增殖

Inhibition of Trifluoperazine on BGC-823 Cell by Inducing p16^(INK4a)

为探索三氟拉嗪(trifluoperazine,TFP)抗肿瘤作用机制,对胃癌BGC-823细胞进行TFP(5、10μmol/L)处理后,利用计数法、BrdU脉冲标记法、Western印迹等方法从细胞形态、细胞增殖、S期细胞百分比以及相关因子表达水平等方面进行分析.结果显示,TFP处理后,细胞形态发生明显改变,细胞增殖受到明显抑制且呈时间计量效应关系;S期细胞比例下降;p16INK4a表达水平升高.为进一步研究TFP诱导p16INK4a表达的分子机制,本实验采用插入p16INK4a启动子片段及荧光素酶报告系统的载体pGL3-Basic-p16INK4a(-967～-165 bp),研究了TFP在转录水平对p16INK4a启动子活性的影响.结果表明,TFP能够提高p16INK4a的启动子活性.上述结果提示,TFP通过诱导p16INK4a表达抑制BGC-823细胞增殖.

To investigate the molecular mechanisms of the anti-tumor effect of trifluoperazine（TFP）,the gastric cancer cell BGC-823 was treated with TFP（5,10 μmol/L） to analyze the cell morphology,proliferation,percentage of cells in S phase and the expression level of relative factors through counting,BrdU pulse labeling,and Western blotting.The results showed that TFP inhibited the cell proliferation in a dose-and time-dependent manner and induced a huge change in morphology.The percentage of cells in S phase was decreased by TFP and the expression level of p16 INK4a was increased dramatically after TFP treatment.We used luciferase reporter pGL3-Basic-p16 INK4a（-967 ～-165 bp） to further investigate the transcriptional effect of TFP on p16 INK4a.The results showed that TFP activated the promoter of p16 INK4a.These results indicate that TFP may positively regulate p16 INK4a to inhibit the proliferation of BGC-823 cell.