Vk3对宫颈癌Hela细胞mTOR信号转导蛋白基因表达的影响

Effect of Vk3 on gene expression of mTOR signaling pathway in cervical carcinoma Hela cells

目的:观察维生素K3(Vk3)对Hela细胞雷帕霉素靶蛋白(mTOR)及其底物p70S6激酶(p70S6K)α1、α2、β1、β2和细胞周期素D1(cyclinD1)mRNA表达的变化及N-乙酰半胱氨酸(NAC)对mTOR及其底物的影响,探讨mTOR信号转导蛋白在人宫颈癌的发生、发展过程中的作用。方法:实验分为对照组(Con)、Vk3处理组(Vk3)、Vk3与NAC联合作用(Vk3+NAC)组及NAC单独应用组(NAC),MTT法检测各组Hela细胞存活率,逆转录聚合酶链反应(RT-PCR)方法检测各组Hela细胞中mTOR、p70S6K-α1、p70S6K-α2、p70S6K-β1、p70S6K-β2及cyclinD1mRNA表达。结果:与Con组比较,Vk3组Hela细胞存活率显著降低(P<0.01),mTOR、p70S6K-α1、p70S6K-α2、p70S6K-β1、p70S6K-β2及cyclin D1 mRNA表达量均明显下降(P<0.05);而与Vk3组比较,Vk3+NAC组Hela细胞存活率显著增加(P<0.01),上述指标mRNA表达量均明显增加(P<0.05)。结论:Vk3通过其氧化应激作用抑制Hela细胞的增殖,其机制与降低mTOR信号转导蛋白及cyclinD1的基因表达有关。

Objective To explore the role of mammalian target of rapamycin （mTOR） in the Hela cells injury induced by vitamin K （Vk3） through observation of mTOR, p70S6K-α1, p70S6K-α2, pTOS6K-β1, p70S6K-β2 as well as cyclin D1 mRNA expression and the effects of N-acetyl cysteine （NAC） on mTOR, p70S6K-α1, p70S6K-α2, p70S6K-β1, p70S6K-β2 as well as eyclin D1 mRNA. Methods Hela cells were randomly divided into control group, Vk3 group, Vk3＋NAC group and NAC group, respectively. The survival rate of Hela cells was determined by MTT. The mTOR, pTOS6K-α1, pTOS6K-α2, pTOS6K-β1, p70S6K-β2 and cyclin D1 mRNA expressions in Hela cells were assayed by using RT-PCR. Results Compared with control group, the survival rate of Hela cells in Vk3 group was decreased （P〈0.01）, the expressions of mTOR, p70S6K-α1, p70S6K-α2, p70S6K-β1, p70S6K-β2 and cyclin D1 mRNA were decreased （P〈0.05）. Compared with Vk3 group, the survival rate in Vk3＋NAC group was increased （P〈0.01）, and the mRNA expressions of those genes mentioned above were also increased （P〈0.05）. Conclusion Vk3 can inhibit Hela cell proliferation through oxidative stress, which might be correlated with down-regulation of the genes expressions of mTOR signal pathway and cyclin D1.