雷公藤内酯醇通过Nrf2/ARE信号通路调控淋巴瘤细胞增殖、凋亡的机制研究

Study on mechanism of triptolide regulating proliferation and apoptosis of lymphoma cells through Nrf2/ARE signaling pathway

目的:探讨雷公藤内酯醇(TPL)对人源套细胞淋巴瘤细胞JeKo-1增殖、凋亡的影响,以及Nrf2/ARE信号通路的可能作用。方法:常规条件培养JeKo-1细胞,不同浓度TPL(0、25、50、100 ng/ml)处理细胞,MTT检测细胞活力,流式细胞术检测细胞凋亡率;Western blot检测Ki67、Bcl-2、Bax、Nrf2、NQO-1和HO-1蛋白表达。qRT-PCR和Western blot检测TPL对转染Nrf2 pcDNA的mRNA和蛋白表达的影响,观察TPL对Nrf2/ARE信号通路增殖和凋亡的作用。结果:随着TPL浓度增加,TPL明显抑制JeKo-1细胞活力,促进细胞凋亡,下调Ki67、Bcl-2、Nrf2、NQO-1和HO-1蛋白表达,上调Bax蛋白表达,且呈剂量依赖性。与TPL+vector组相比,TPL+Nrf2 pcDNA组Nrf2 mRNA和蛋白表达显著增加,细胞活力显著提高,凋亡率显著降低,Ki67、Bcl-2蛋白表达上调,Bax蛋白表达下调。结论:TPL可明显抑制JeKo-1细胞增殖,促进细胞凋亡,其作用机制可能与Nrf2/ARE信号通路有关。

Objective:To investigate effect of triptolide(TPL)on proliferation and apoptosis of human mantle cell lymphoma cell JeKo-1,and possible role of Nrf2/ARE signaling pathway.Methods:JeKo-1 cells were cultured under routine conditions,and cells were treated with different concentrations of TPL(0,25,50,100 ng/ml).Cell viability was detected by MTT.Apoptosis rate was detected by flow cytometry,and expressions of Ki67,Bcl-2,Bax,Nrf2,NQO-1 and HO-1 proteins were detected by Western blot.qRT-PCR and Western blot to detect effect of pcDNA transfected by TPL on mRNA and protein expressions of Nrf2,and to observe effect of TPL on proliferation and apoptosis of Nrf2/ARE signal pathway.Results:With increasing of TPL concentration,TPL significantly inhibited viability of JeKo-1 cells,promoted cell apoptosis,down-regulated expressions of Ki67,Bcl-2,Nrf2,NQO-1 and HO-1,and up-regulated expression of Bax protein,which was in a dose-dependent manner.Compared with TPL+vector group,expressions of Nrf2 mRNA and protein in TPL+Nrf2 pcDNA group were significantly increased,which significantly promoted cell viability,significantly inhibited rate of apoptosis,and up-regulated Ki67 and Bcl-2 protein expression,and down-regulated Bax protein expression.Conclusion:TPL can significantly inhibit proliferation of JeKo-1 cells and promote cell apoptosis,whose mechanism may be related to Nrf2/ARE signaling pathway.