普洱茶水提取物体外抑制舌鳞癌Tca8113细胞增殖并诱导凋亡的研究

Study of Pu-erh tea extract on inhibiting proliferation and inducing apoptosis of tongue squamous cell carcinoma Tca8113 cell in vitro

目的 研究普洱茶水提取物（Pu-erh tea extract,PTE）对舌鳞癌Tca8113细胞的抑制作用及其作用机制.方法 采用四甲基偶氮唑盐比色法检测PTE对Tca8113细胞和牙周膜成纤维细胞生长和增殖的抑制作用,用倒置相差显微镜观察PTE作用后细胞的形态变化,利用流式细胞术分析PTE对舌鳞癌细胞的作用机制.实验数据采用GraphPad Prism 5.01软件进行统计分析,应用单因素方差分析对数据进行比较.结果 在PTE作用时间相同的情况下,随着其浓度的增加,Tca8113细胞存活率明显降低,当PTE浓度为125.00 μg/ml时,细胞存活率下降最为显著（F=1 283,P＜0.000 1）.而当PTE浓度相同的情况下,随着作用时间的增加,Tca8113细胞存活率也随之降低,并且在作用12h时差异最显著（F=111.6,P＜0.000 1）.PTE对正常牙周膜成纤维细胞无明显抑制作用.显微镜观察发现,舌鳞癌细胞经PTE处理后出现皱缩、变圆、脱离贴壁状态;而牙周膜成纤维细胞形态无变化.流式细胞术检测发现,舌鳞癌细胞经125.00 μg/ml的PTE作用12h后,细胞凋亡率为27.65％±0.47％.结论 PTE能通过诱导细胞凋亡抑制舌鳞癌Tca8113细胞增殖,提示其具有防治舌鳞癌的作用.

Objective To study inhibition effect of Pu-erh tea extract （PTE） on tongue squamous cell carcinoma Tca8113 cell and its mechanism.Methods The inhibition on proliferation and growth of Tca8113 cell and periodontal ligament fibroblast （PDL） was detected with methyl-thiazol-tetrazolium method.The morphological changes of cells were observed under inverted phase contrast microscope.The mechanism of inhibition was studied using flow cytometry.The data were analyzed with GraphPad Prism 5.01 software and compared by one-way ANOVA.Results Tca8113 cell viability decreased clearly with the increase of PTE concentration.When the concentration of PTE was 125.00 μg/ml,the most significant decrease was observed （F=1 283,P〈0.000 1）.Tca8113 cell viability decreased with the increase of time with the most significant difference at 12 h （F=111.6,P〈0.000 1）.However,the growth and proliferation of PDL were not influenced by PTE.Shrinkage,roundness and detachment were observed in Tca8113 cell but not in PDL under microscope.Apoptosis rate was 27.65% ±0.47% after Tca8113 cells were treated with 125.00 μg/ml PTE for 12 h.Conclusion PTE can inhibit Tca8113 cell proliferation by inducing apoptosis,suggesting that it could prevent or treat tongue squamous cell carcinoma.