Geng Ying-Hua, Wu Wen-Juan, Li Jun, Zhou Li-Li, Yang Yan-Li
Department of Hematology, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, Anhui Province, China,E-mail:
Department of Biochemistry and Molecular Biology, Bengbu Medical College, Bengbu 233030, Anhui Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020 Feb;28(1):110-118. doi: 10.19746/j.cnki.issn.1009-2137.2020.01.019.
To investigate the proliferation inhibition and pro-apoptosis effect of LY294002 (PI3K/AKT inhibtor) combined with daunorubicin (DNR) on the chronic myeloid leurenia cell line K562 and its possible mechanisms.
The effect of LY294002 and DNR on the proliferation of K562 cells in different treating time and concentration were measured by MTT assay. The cell cycle was determined by flow cytometry, the mRNA and protein expression of SKP2 , P27, BCL-2 and BAX were determined by RT-PCR and Western blot.
LY294002 and DNR were able to inhibit the growth of K562 cells and promote apoptosis in time- and concentration-dependent manner (P＜0.05), both the cell proliferation-inhibiting rate and apoptosis rate in combination therapy group were higher than that in DNR-monotherapy group (P＜0.05). After K562 cells treated by LY294002 combined DNR for 36 h, the cells were statistically significantly reduced in G/M phase (P＜0.05), as compared with control group and DNR group. Compared with DNR group, the cell level of G/G phase rased (P＜0.05) and cell level of S phase decreased (P＞0.05). Compared with DNR group, the expresson of SKP2 and BCL-2 mRNA decreased, and the expression of P27 mRNA increased in the combination therapy group (P＜0.05). The expression of BAX mRNA was not significantly different between different groups. The same result was found in the protein expression.
LY294002 has the sensibilizative effect on DNR chemotherapy, which may be relative with blocking the cell cycle and inducing cell apoptosis.
探讨LY294002（PI3K/AKT抑制剂）联合柔红霉素（DNR）对慢性髓性白血病细胞系K562的增殖抑制及促凋亡作用及其可能机制。
采用MTT法检测LY294002和DNR在不同作用时间和浓度下对K562细胞增殖的影响。通过流式细胞术检测细胞周期，采用RT-PCR和Western blot检测SKP2、P27、BCL-2和BAX的mRNA及蛋白表达。
LY294002和DNR均能抑制K562细胞生长并促进其凋亡，呈时间和浓度依赖性（P＜0.05），联合治疗组的细胞增殖抑制率和凋亡率均高于DNR单药治疗组（P＜0.05）。LY294002联合DNR作用K562细胞36 h后，与对照组和DNR组相比，G/M期细胞比例统计学显著降低（P＜0.05）。与DNR组相比，联合治疗组G/G期细胞比例升高（P＜0.05），S期细胞比例降低（P＞0.05）。联合治疗组与DNR组相比，SKP2和BCL-2 mRNA表达降低，P27 mRNA表达升高（P＜0.05）。不同组间BAX mRNA表达无显著差异。蛋白表达结果相同。
LY294002对DNR化疗具有增敏作用，可能与阻滞细胞周期及诱导细胞凋亡有关。
