姜黄素通过TGF-β1信号通路抑制人肺泡横纹肌肉瘤细胞株PLA-802的机制研究

Inhibitory effect of Curcumin on human alveolar rhabdomyosarcoma cell line PLA-802 through inhibiting TGF-β1 signaling pathway

摘要:

目的：观察姜黄素对人横纹肌肉瘤(Rhabdomyosarcoma,RMS)细胞株PLA-802的抑制作用，以及对细胞内TGF-β1/Smad 4的表达的影响，探索姜黄素抑制RMS细胞生长的机制。方法：体外培养人RMS细胞株PLA-802，并用不同浓度的姜黄素作用不同的时间。用MTT检测姜黄素对PLA-802细胞生长情况的影响，用流式细胞术检测细胞周期的变化情况，RT-PCR和Western blot分别检测细胞内TGF-β1和Smad 4的mRNA和蛋白水平的表达。结果：MTT结果显示姜黄素显著降低了PLA-802细胞的存活率（P<0.05)。流式细胞结果表明姜黄素明显降低了S期而增加了G1期(P<0.05)。而TGF-β1和其下游因子Smad4的mRNA和蛋白水平的表达也明显受到姜黄素的抑制，且这种抑制作用呈浓度-时间依赖性(P<0.05)。结论：姜黄素发挥其抑制PLA-802细胞的作用可能是通过抑制TGF-β1信号通路，这将为临床治疗肺泡RMS提供新的思路。

Abstract:

Objective:To observe the inhibitory effect of Curcumin on human alveolar rhabdomyosarcoma cell line PLA-802 in vitro, to detect the expressions of TGF-beta1 and Smad 4, and to explore its potential mechanisms. Methods：Human alveolar rhabdomyosarcoma cell line PLA-802 was cultured and treated with Curcumin at different concentration and at different time points. The effect of Curcumin on PLA-802 cell proliferation was studied by means of MTT; the cell cycle was detected with flow cytometry; RT-PCR and western blot were used to detect the expressions of TGF-β1 and Smad 4 at mRNA and protein levels. Results：The viability of PLA-802 cells treated with Curcumin was obviously decreased (P<0.05). Analysis of the cell cycle revealed that Curcumin induced a significant decrease in cells in the S phase and an increase in cells in the G1 phase (P<0.05). Furthermore, the expressions of TGF-β1 and its downstream factor Smad 4 at mRNA and protein levels were inhibited by Curcumin in a concentration and time dependent manner (P<0.05). Conclusion:Curcumin perhaps plays its inhibitory effects on PLA-802 cells through inhibiting the the activity of TGF-β1 signaling pathway, which will provide an important therapeutic potential for treating human aleovar rhabdomyosarcoma.