汉防己甲素抑制口腔癌细胞增殖与β-catenin表达的研究

Study of the cellular growth inhibited by tetrandrine with the expression of β-catenin in oral squamous cell carcinoma cell line

目的研究汉防己甲素(tetrandrine,TET)对口腔癌细胞Tca8113的作用及其对β-连环蛋白(β-catenin)蛋白表达的影响。方法应用CCK-8法检测不同浓度TET对Tca8113细胞作用不同时间后的影响;应用Western blot法检测TET作用前后β-catenin蛋白表达的变化情况;应用细胞免疫荧光法检测TET作用前后β-catenin蛋白的分布。结果随着TET浓度的增加,其对Tca8113的抑制率也不断增加(F=819.90,P<0.05),半数有效抑制浓度(half maximal inhibitory concentration,IC50)为64μmol/L;同时TET对Tca8113细胞的抑制呈时间依赖性。Western blot结果显示,在TET作用后β-catenin蛋白的表达下调,差异具有统计学意义(t=6.48,P<0.05)。细胞免疫荧光法检测结果显示,在空白组中β-catenin蛋白主要表达在细胞核内,而在TET作用后β-catenin蛋白主要表达在细胞膜和细胞质中,呈散在分布,细胞核内表达较少。结论 TET能显著抑制口腔癌细胞Tca8113的增殖,同时可影响β-catenin蛋白在Tca8113细胞内的分布。

Objective To study the effect of tetrandrine (TET) on oral squamous cell carcinoma cell line Tca8113 and its influence against the expression of β-catenin. Methods CCK-8 method was used to detect the effect of TET on Tca8113 by different dose and time. Western blot assay was used to detect the expression of β-catenin after treatment by TET meanwhile the distribution of the β-catenin treated by TET was detected by the immunofluorescence method.Results With the increasing dose of TET, the inhibition rate against Tca8113 was raising (P < 0.05). The half maxi-mal inhibitory concentration (IC50) value of TET against Tca8113 was 64 μmol/L. With this concentration, the inhibition rate against Tca8113 exhibited a time-dependent manner. The results of western blot revealed that the expression of β-catenin was significantly inhibited after treatment by TET (P < 0.05). The results of immunofluorescence methodshowed that the β-catenin protein mainly located at nucleus in Tca8113, while after treatment by TET, the β-cateninprotein mainly located at cellular membrane and cytoplasm in a diffused distribution manner. Conclusion Our data suggest that TET can inhibit the cell growth of oral squamous cell carcinoma, and influence the cellular distribution of β-catenin protein in tumor cells.