姜黄素调控mTOR信号通路对淋巴瘤细胞DOHH-2增殖和凋亡的作用研究

Effect of Curcumin on the Proliferation and Apoptosis of Lymphoma Cell DOHH-2 by Regulating the mTOR Signaling Pathway

目的:探讨姜黄素通过调控哺乳动物雷帕霉素靶蛋白(mTOR)信号通路对淋巴瘤细胞DOHH-2增殖、凋亡的作用及机制。方法:采用四唑盐(MTT)法筛检姜黄素干预的淋巴瘤细胞半致死剂量(LD50)条件;将对数生长期DOHH-2细胞分为空白组、姜黄素组、MHY1485组及姜黄素+MHY组,姜黄素组以LD50的姜黄素干预,MHY1485组用mTOR通路激活剂MHY1485干预,姜黄素+MHY组同时用LD50的姜黄素及MHY1485干预。MTT法和流式细胞术检测4组细胞增殖、凋亡情况,蛋白免疫印迹(Western blot)法检测磷酸化-mTOR(p-mTOR)、磷酸化p70核糖体蛋白S6激酶(p-p70S6K)、Caspase3蛋白表达情况。结果:姜黄素可呈剂量和时间依赖性降低DOHH-2细胞存活率,LD50条件为20μmol/L的姜黄素干预48 h;20μmol/L的姜黄素干预48 h时,细胞存活率姜黄素组低于空白组,姜黄素+MHY组低于MHY1485组,姜黄素+MHY组高于姜黄素组,差异均有统计学意义(P<0.05);与空白组和姜黄素+MHY组比较,姜黄素组细胞凋亡率及Caspase3蛋白相对表达量较高,MHY1485组较低(P<0.05);与空白组和姜黄素+MHY组比较,姜黄素组p-mTOR、p-p70S6K蛋白相对表达量较低,MHY1485组较高(P<0.05)。结论:姜黄素可有效抑制淋巴瘤细胞DOHH-2增殖,促进其凋亡,其机制可能与抑制mTOR信号通路的激活有关。

Objective:To investigate the effect of curcumin on the proliferation and apoptosis of lymphoma cells DOHH-2 by regulating the mammalian target of rapamycin(mTOR)signaling pathway.Methods:The median lethal dose(LD50)condition of curcumin intervention was screened by MTT method.The log phase DOHH-2 cells were divided into blank group,curcumin group,MHY1485 group and curcumin+MHY group.Curcumin group was treated with LD50 curcumin,MHY1485 group was treated with mTOR pathway activator MHY1485,curcumin+MHY group was treated with LD50 curcumin and MHY1485 at the same time.The proliferation and apoptosis of the four groups were detected by MTT method and flow cytometry.The expressions of phosphorylated-mTOR(p-mTOR),phosphorylated p70 ribosomal protein S6 kinase(p-p70S6K),and Caspase3 protein were detected by Western blot.Results:Curcumin can reduce the survival rate of DOHH-2 cells in a dose and time-dependent manner.The LD50 condition is 20μmol/L curcumin intervention for 48 h.When 20μmol/L curcumin intervenes for 48 hours,the cell survival rate of curcumin group was lower than that of blank group,curcumin+MHY group was lower than MHY1485 group,and curcumin+MHY group was higher than curcumin group with statistical differences(P<0.05).Compared with the blank group and the curcumin+MHY group,the apoptosis rate and the relative expression of Caspase3 protein in the curcumin group were higher,and the MHY1485 group was lower(P<0.05).Compared with the blank group and the curcumin+MHY group,the relative expressions of p-mTOR and p-p70S6K protein in the curcumin group were lower,and those in the MHY1485 group were higher(P<0.05).Conclusions:Curcumin can effectively inhibit the proliferation of lymphoma cells DOHH-2 and promote their apoptosis.Its regulatory mechanism may be related to the inhibition of the activation of mTOR signaling pathway.