开口箭皂苷对人甲状腺髓样癌TT细胞的抑制作用及对Notch1的影响

Inhibitory effects of total saponins of tupistra chinensis in human medullary thyroid carcinoma TT cells and its effect on expression of Notch1

关键词:  开口箭  甲状腺髓样癌  Notch1  DAPT

Key words:  Tupistra chinensis Baker  medullary thyroid carcinoma  Notch1  DAPT

摘要:

[目的] 研究开口箭皂苷（TST）对人甲状腺髓样癌TT细胞（MTC-TT）的抑制作用及对Notch1表达水平的影响，探讨TST抗甲状腺髓样癌的作用机制。[方法] 将不同浓度的TST作用于MTC-TT细胞，CCK8法测TST对MTC-TT细胞的增殖抑制率并计算出24、48、72 h时半数抑制浓度（IC₅₀）；流式细胞术检测细胞周期；Western blot检测各组Notch 1的蛋白表达，RT-PCR检测各组Notch 1的基因表达；将TST+DAPT（Notch1抑制剂）作用于MTC-TT细胞48 h，Western blot测Notch1表达，CCK8法测细胞增殖抑制率。[结果] TST对MTC-TT细胞的24 h、48 h和72 h的IC₅₀值分别为59.01、31.54和31.63 μg/mL，与对照组相比，TST对MTC-TT细胞增殖有抑制作用（P<0.01），细胞周期阻滞在G₂/M期（P<0.05，P<0.01）；实验组Notch1的蛋白和mRNA表达水平相比于对照组均明显升高（P<0.05，P<0.01），表达呈浓度依赖性（P<0.01）；与TST组相比，TST+DAPT组Notch1蛋白表达显著下降（P<0.01），细胞增殖抑制率显著下降（P<0.01）。[结论] TST可抑制MTC-TT细胞增殖，诱导MTC-TT细胞凋亡，该作用可能与上调Notch 1表达水平有关。

Abstract:

[Objective] To study the inhibitory effects of total saponins of Tupistra chinensis Baker (TST) in human medullary thyroid carcinoma TT (MTC-TT) cells and its effect on expression of Notch1 and investigate its anticancer mechanism.[Methods] MTC-TT cells were treated with different density of TST, the cell proliferation inhibition rate and IC₅₀ at 24 h, 48 h, 72 h were evaluated by CCK8;the disposition of MTC-TT cells cycle was detected by flow cytometry. The expression of Notch1 protein were detected by western blot and the mRNA were detected by Real-time PCR; in addition, the expression of Notch1 protein were detected by western blot and the cell proliferation inhibition rate was detected by CCK8 after TST and DAPT (Notch1 inhibitor) treated 48h.[Results] The IC₅₀ at 24 h, 48 h,72h was 59.01, 31.54 and 31.63 μg/mL respectively. Compared to control group, TST has inhibitory effects on MTC-TT cells (P<0.01), cell cycle were arrested in G₂/M phase (P<0.05, P<0.01). Compared to control group,the expression of Notch1 protein and mRNA in experience group increased significantly after treatments (P<0.05, P<0.01), the high dose group increased more significantly in a concentration-dependent manner (P<0.01). Compared to TST group, the expression of Notch1 protein in TST +DAPT group decreased significantly (P<0.01), the cell proliferation inhibition rate decreased,either (P<0.01).[Conclusion] TST could inhibit the proliferation and cell cycle of MTC-TT cells by probable up-regulation of the Notch1 expression.