苦参碱对Raji细胞增殖及Notch信号通路的影响

Matrine suppresses proliferation of Raji cells via inhibition of Notch signaling pathway

目的探讨苦参碱对淋巴瘤细胞株Raji细胞增殖及Notch信号通路的影响。方法试验分为5组:对照组(0 g/L)和4个苦参碱组(0.5、0.75、1.0、1.5 g/L),分别于苦参碱作用于Raji细胞24、48、72 h后,采用MTT法检测细胞增殖,流式细胞仪检测细胞凋亡及周期分布,RT-PCR法及Western blot法检测Notch1受体及下游靶基因Hes1 m RNA转录及蛋白的表达。结果不同浓度苦参碱均有抑制Raji细胞增殖的作用,且随药物浓度增加及作用时间延长,抑制作用更明显;苦参碱可明显诱导Raji细胞凋亡;经苦参碱1.0 g/L处理48 h后,G1期细胞减少(P<0.05),S期细胞增多(P<0.05),Notch1受体及下游靶基因Hes1 m RNA转录水平及蛋白表达水平均明显下调(P<0.01)。结论苦参碱可抑制淋巴瘤细胞株Raji细胞增殖,且呈浓度-时间依赖性,并诱导细胞凋亡,使细胞周期停留于S期,可能通过直接或间接下调Notch信号通路相关基因表达而实现。

Objective To investigate the effects of matrine on proliferation of lymphoma Raji cells and Notch signaling pathway. Methods Raji cells were divided into five groups. The cells in control group were untreated, while those in four test groups were treated with matrine at concentrations of 0. 5, 0. 75, 1. 0 and 1. 5 g/L respectively. The proliferation levels of cells 24, 48 and 72 h after treatment were determined by MTT assay, while the apoptosis and cell cycle by flow cytometry, and the m RNA transcription and protein expression levels of Notch1 receptor and target gene Hes1 by RT-PCR and Western blot respectively. Results Matrine inhibited the proliferation of Raji cells in a dose-and time-dependent pattern, which induced the cell apoptosis significantly. After treatment with 1. 0 g/L matrine for 48 h, the cells in G1 phase decreased（P〈 0. 05） while those in S phase increased（P〈 0. 05）. Both the m RNA transcription and protein expression levels of Notch1 receptor and target gene Hes1 decreased significantly（P〈 0. 01）. Conclusion Matrine may inhibit the proliferation of Raji cells in a dose-and time-dependent pattern, induce the cell apoptosis, and arrest the cell cycle in S phase by a possible mechanism of down-regulating the expression of Notch signal pathway related genes directly or indirectly.