关于细胞毒药物下调细胞外调节蛋白激酶与抑制肝癌smf7721细胞增殖.pdf

关于细胞毒药物下调细胞外调节蛋白激酶与抑制肝癌SMF7721 细胞增殖 【关键词】肝肿瘤 关键词: 肝肿瘤;蛋白激酶类;细胞周期;细胞分裂 摘 要:目的研究三种细胞毒药物（三尖杉酯碱，长春新碱，足叶已甙）抑 制肝癌SMF7721 细胞增殖、阻滞细胞周期过程中，细胞外调节蛋白激酶 （extracelluar regulated protein kinases ERK1/2）磷酸化蛋白表达水平的 变化.探讨细胞毒药物杀伤肝癌SMF7721 的作用机制. 方法 采用MTT 法测定增 殖抑制率，应用流式细胞术检测细胞周期，应用Western blot 方法观察 ERK1/2 磷酸化蛋白表达水平的变化. 结果 三种细胞毒药物（三尖杉酯碱，长 春新碱，足叶已甙）均能不同程度的抑制SMF7721 细胞增殖，抑制率分别为 （28±8）%，（25±16）%和（24±11）%;三尖杉酯碱可使SMF7721 细胞周期阻 滞于G1 期，长春新碱可使其阻抑于S 期和G2/M 期，Vp16 则使之受抑于G2/M 期.受这三种细胞毒药物作用后，肝癌SMF7721 细胞内处于活化状态的磷酸化 ERK1/2 蛋白表达水平下降.ERK1 分别为对照组的43%，32%，72%，ERK2 分别为 对照组的29%，20%，61%. 结论 ERK 通路可能是细胞毒药物杀伤肝癌细胞 SMF7721 的作用机制之一. Keywords:liver neoplasms;protein kinases;cell cycle;cell pision Abstract:AIM To study the change of phosphorylated （ac-tivated） ERK1and ERK2protein expression in the course of inhibiting hepatocarcinomatous cell proliferation and arresting cell cycle progression by three kinds of cytotoxic drugs，Har-ringtonine （HRT ）， Vincristine （VCR）and Etoposide （Vp16），and to discuss the killing mechanisms of cytotoxic drugs to hepato-carcinomatous cell， SMF7721.METHODS Proliferation inhibition rate was detected by MTT method.The cell cycle was analyzed by flow cytometry.The change of the phosphorylated ERK1/2protein expression was ob-served by Western blot method.RESULTS Three kinds of cytotoxic drugs （HRT，VCR and Vp16） inhibited hepatocar-cinomatous cell proliferation to some extent.The inhibition rate was （28±8）%，（25±16）%and （24±11）%respective- ly.The cell cycle was blocked in G1phase with HRT.The cell cycle was blocked in S phase and G2/M phase with VCR.And the cell cycle was blocked in G2/M phase with Vp16.In experimental groups，the contents of phosphorylated ERK1and ERK2protein which was active in hepatocarcinomatous cell SMF7721obviously declined.With the effects of three cytotoxic drugs，the content of ERK1was43%， 32%and72%respectively;the content of ERK2was29%， 20%and61%respectively.CONCLUSION It mig