替莫唑胺对小细胞肺癌H446细胞的凋亡诱导作用

Inductive effect of temozolomide on small cell lung cancer H446 cell apoptosis

目的探讨替莫唑胺对小细胞肺癌(SCLC)细胞H446的凋亡诱导作用及具体分子生物学机制。方法通过MTT法检测替莫唑胺对H446细胞活力的影响;流式细胞术检测替莫唑胺对H446细胞周期的影响;Western blot分析磷脂酰肌醇-3-激酶(PI3K)/AKT通路的激活状况及其下游靶基因细胞周期蛋白B1(Cyclin B1),细胞分裂周期基因2(Cdc2),Bax,Bcl-2和生存素(Survivin)的表达。结果替莫唑胺(50,100,200μmol/L)可显著抑制细胞的活力,并且在48h抑制程度最高,并使细胞周期阻滞在G2/M期。Western blot结果显示替莫唑胺能抑制PI3K表达及Akt磷酸化,进而下调Cyclin B1,Cdc2,Bcl-2和Survivin的表达,上调Bax的表达。结论替莫唑胺可通过阻断PI3K/AKT信号通路来诱导SCLC细胞H446凋亡。

Objective To explore the effect of temozolomide on apoptosis and molecular mechanism in small cell lung cancer （SCLC） cell H446. Methods The effect of temozolomide on the viability of H446 cell was measured by MTT assay.The effect of temozolomide on cell cycle was detected by flow cytometry.The activation of phosphatidyl inositol 3-kinase （PI3K）/AKT signaling pathway and expression level of downstream target genes （Cyclin B1）, cell division cycle 2 （Cdc2）, Bax, Bcl-2 and Survivin were detected by western blot.Results Temozolomide （50, 100, 200 μmol/L） could inhibit H446 cell viability, and the inhibitory rate was highest at 48 h.Moreover, temozolomide made H446 cell cycle arrested in G2 phase.Western blott showed the expression of PI3K, Cyclin B1, Cdc2, Bcl-2, Survivin and the phosphorylation of AKT were reduced, but the expression of Bax were increased by temozolomide.Conclusion Temozolomide could induce SCLC cell H446 apoptosis via blocking PI3K/AKT signal pathway.