NVP-BEZ235抑制CD34+CD38-急性髓系白血病干细胞的增殖和集落形成

摘要

本研究旨在探讨PI3 K/mTOR信号系统双靶点抑制剂——NVP-BEZ235对CD34+ CD38-髓系白血病干细胞增殖、细胞周期和集落形成能力的影响.用流式细胞术检测急性髓系白血病KG1a细胞表面CD34和CD38的表达;应用台盼蓝染色细胞计数法检测不同浓度NVP-BEZ235对KG1a细胞增殖的影响,PI染色流式细胞术检测NVP-BEZ235对KG1a细胞周期的影响,持续用软琼脂集落形成实验检测不同浓度NVP-BEZ235对KG1a细胞集落形成细胞的影响.结果表明,急性髓系白血病KG1a细胞中CD34+ CD38-占(98.02±0.72)％,NVP-BEZ235(0.125-1μmol/L)对KG1a细胞增殖抑制呈现时间和剂量依赖(P＜0.05),其24和48 h的IC50值分别为0.597和0.102 μmol/L.0.5 μmol/L的NVP-BEZ235作用24 h后,Go/G1期KG1a细胞比例达(83.2±3.80)％,较对照组(43.47±9.60)％明显增高(P＜0.05).2500个细胞在NVP-BEZ235 (0-1 μmol/L)持续作用下14 d和21 d而形成的集落数分别由375.67±21.46、706.33±87.31降至0(P ＜0.05).结论:NVP-BEZ235能抑制CD34+ CD38-髓系白血病干细胞增殖和集落形成.%This study was aimed to explore the effect of NVP-BEZ235, a dual phosphatidylinositol 3-kinase/ mammalian target of rapamycin inhibitor, on proliferation, cell cycle and colony forming capability of CD34+ CD38-human acute myeloid leukemia (AML) KGla cells. Flow cytometry was used to detect expresstion of CD34 and CD38 on the surface of human AML KG1 a cells; Trypan blue assay was used to analyze the effect of NVP-BEZ235 at various concentrations on proliferation of KG1 a cells; flow cytometry was performed to examine the cell cycle of KG1 a cells after NVP-BEZ235 treatment; Soft agar colony-forming experiment was used to detect the colony forming ability of KG1 a cells treated with NVP-BEZ235 at various concentrations. The results indicated that the percentage of CD34+ CD38- AML KGla cells was (98.02 ± 0. 72)%. NVP-BEZ235 (0. 125 - 1 μmol/L) inhibited the proliferation of KG1 a cells in a time-and dose-dependent manner (P < 0.05) and the 50% inhibition concentrations (IC50) at 24 h and 48 h were 0. 597 μmol/L and 0.102 μmol/L, respectively. KGla cells were arrested at G0/G1 phase after treating with 0.5 μmol/L NVP-BEZ235 for 24 h, it was significantly higher than that of control group (83. 2 ± 3. 80) % vs (43.47 ± 9. 60) % (P < 0.05). KG1 a cells treated with NVP-BEZ235 (0-1 μmol/L) for 14 d and 21 d, the number of colony decreased respectively from (375.67 ±21.46) per 2500 KGla cells and (706. 33 ±87. 31) per 2500 KGla cells to 0, with statistical significance (P <0.05). It is concluded that NVP-BEZ235 can inhibit proliferation and colony-forming capability of CD34 + CD38 - human AML KGla cells.