【摘要】 目的观察 AKT 及磷酸化 AKT（pAKT,即 AKT 呈活化状态）)蛋白激酶在三个弥漫性大 B 细胞淋巴瘤（DLBCL）细胞株中的表达状态,并探讨其与 DLBCL 细胞生物学行为的关系以及P13K 抑制剂 LY294002对该瘤细胞生长的影响。方法采用 Western blot 检测 DLBCL 细胞株 ly1、ly8、ly10在有血清和无血清培养条件下 AKT 和 pAKT 的表达状态;用 P13K 抑制剂 LY294002分别作用于三株细胞,观察 pAKT 水平的改变;用流式细胞术结合碘化丙啶（PI）单染分析细胞周期、AnnexinV-异硫氢酸荧光素（FITC）染色检测细胞凋亡、BrdU 法观察细胞增殖状态的改变。结果三株DLBCL 细胞株在不同的培养条件下均显示有 pAKT,在10%FBS 培养组 ly8和 ly10之 pAKT 水平高于无血清培养组;而 ly1相反,在无血清培养组 pAKT 略高于有血清培养组。在有血清培养组,LY294002可明显降低三细胞株细胞的 pAKT 水平,并显示 S 期细胞明显减少（P<0.05）,G₀～G₁期细胞增加,增殖细胞比例显著降低（P<0.05）,以 ly8、ly10细胞尤为明显;但凋亡细胞无明显增加（P>0.05）。结论 AKT 活化与 DLBCL 细胞周期和细胞增殖密切相关;与细胞凋亡无显著相关;LY294002可通过降低 DLBCL 细胞的 pAKT 水平继而抑制瘤细胞生长。更多还原

【Abstract】 Objective To observe the status of AKT and phospho-AKT(pAKT)in three diffuse large B cell lymphoma(DLBCL)cell lines,and to investigate the effects of AKT activation on biologic behavior of DLBCL cells.Methods Three DLBCL cell lines,lyl,ly8 and lyl0 were maintained in 10% FBS or serum free culture medium.The expression of AKT and status of pAKT were detected by Western blotting.LY294002,an inhibitor of PI3K,was used to suppress the level of pAKT.Flow cytometry combined with PI staining,Annexin V-FITC assay and Brdu incorporation assay were used to analyze the parameters of the cell cycle,apoptosis and proliferation respectively.Results There was constitutive activation of AKT in three DLBCL cell lines and the levels of pAKT were altered in the different environments.In 10% FBS culture medium,pAKT was higher than that in serum free culture medium in ly8 and lyl0,however,pAKT in lyl maintained in seium free culture medium was mildly higher than that in 10% FBS culture medium.When the cell lines lyl,ly8,1yl0 were maintained in 10% FBS culture medium, the inhibitor LY294002 suppressed the level of pAKT efficiently in three DLBCL cell lines.The percentage of ceils at S phase and the proliferation index were significantly decreased(P < 0.05)without an increase of apoptosis(P>0.05).Conclusions Activation of AKT may play an important role in the development of DLBCL.It is closely related to the control of cell cycle and proliferation,but is not associated with apoptosis.LY294002 can inhibit cell growth by decreasing the levels of pAKT in DLBCL cell lines.更多还原