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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),G_0～G_1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。
Objective To observe the expression of AKT and phosphorylated AKT (activated by pAKT) AKT) in three diffuse large B cell lymphoma (DLBCL) cell lines and to investigate its relationship with the biological behavior of DLBCL cells And P13K inhibitor LY294002 on the growth of the tumor cells. Methods Western blot was used to detect the expression of lymphatic cell lysates ly1, ly8, and ly10 in serum and serum-free medium, and the effects of P13K inhibitor LY294002 on three cell lines were observed. The changes of pAKT levels were observed by flow cytometry Cell cycle was analyzed by PI staining, apoptosis was detected by Annexin V-fluorescein isothiocyanate (FITC) staining, BrdU assay was used to observe the cell proliferation. Results All three DLBCL cell lines showed pAKT under different culture conditions. The pAKT levels of ly8 and ly10 cells in 10% FBS group were higher than those in serum-free medium group. On the contrary, in lyc1-serum group, pAKT was slightly higher than Serum culture group. In serum-cultured group, LY294002 could significantly reduce the pAKT levels in the three-cell lines and showed a significant decrease in S phase cells (P <0.05), while the cells in G_0 ~ G_1 phase increased significantly (P <0.05) ly8, ly10 cells is more obvious; but no significant increase in apoptotic cells (P> 0.05). Conclusion AKT activation is closely related to the cell cycle and cell proliferation of DLBCL, but not to apoptosis. LY294002 can inhibit the growth of tumor cells by decreasing the pAKT level of DLBCL cells.