雷公藤内酯醇抑制Raji细胞增殖和迁移的体外实验研究

Inhibitory effects of triptolide on cell proliferation and metastasis in Raji cells in vitro

目的:研究雷公藤内酯醇在体外是否具有抗淋巴瘤细胞增殖和迁移的效应。方法:采用四甲基偶氮唑蓝(MTT)比色法检测雷公藤内酯醇对Raji细胞增殖的作用;采用Annexin V/PI双标流式细胞术检测雷公藤内酯醇对Raji细胞凋亡的影响;采用流式细胞仪检测雷公藤内酯醇对Raji细胞表面CXCR4受体表达的影响;并采用Transwell微孔隔离室迁移实验观察雷公藤内酯醇对Raji细胞体外迁移作用的影响。结果:①雷公藤内酯醇能以时间和剂量依赖方式抑制Raji细胞的增殖,24 h和36 h的IC50值分别为43.06 nmol/L和25.08 nmol/L。②随雷公藤内酯醇药物浓度的增加和作用时间的延长,Raji细胞凋亡率逐渐增高,且呈时效和量效关系。③Raji细胞经不同浓度雷公藤内酯醇(12.5、25、50 nmol/L)处理后,CXCR4表达率(34.66%±4.34%、23.74%±1.87%、18.10%±1.18%)明显低于对照组(72.47%±7.28%,P<0.01),此抑制效应呈剂量依赖性;④Transwell趋化活性分析显示雷公藤内酯醇能抑制rhSDF-1α对Raji细胞的趋化作用,此抑制效应也呈量效关系。结论:雷公藤内酯醇具有抗淋巴瘤细胞增殖和诱导凋亡的效应,并能阻断Raji细胞的体外迁移,其机制与其对SDF-1/CXCR4生物学轴的抑制效应有关。

AIM： To investigate the inhibitory effects of triptolide on cell proliferation and metastasis in Burkitt＇s lympboma cell line Raji cells. METHODS： The effects of triptolide on the growth of Raji cells were studied by 3 - （4,5 - Dimethyl -2 - thiazolyl ） -2,5 - diphenyl - 2H - tetrazolium（MTT） assay. The effects of triptolide on the cell apoptosis of Raji cells were detected by using Annexin V/PI double - labled cytometry. The effects of triptolide on CXCR4 expression on Raji cells were studied by flow cytometric analysis. Chemotaxis assays were performed to observe the effects of triptolide on migration of Raji cells towards recombinant human SDF - 1α （rhSDF - 1α）in vitro. RESULTS： Triptolide inhibited the proliferation of Raji cells in a dose - and time - dependent way with a 24 h IC50 value of 43.06 nmol/L and a 36 h IC50 value of 25.08 nmol/L. Following the treatment of triptolide, the cell apoptosis rate was increased as the treatment concentration increased and the culture time extended. The effects were dose - and time - dependent. Triptolide could downregulate the expression of CXCR4 on Raji cells in a dose -dependent manner. Moreover, chemotaxis assay suggested that triptolide could block the migration of Raji cells to rhSDF - 1α in vitro, and the inhibition was dose - dependent. CONCLUSION： Triptolide could inhibit the cell proliferation and induce the cell apoptosis of Raji cells. Furthermore, it could block the cell metastasis of Raji cells in vitro and the underlying mechanism might be related to the inhibition of the SDF - 1/CXCR4 axis.