Yu Wen-juan, Rao Qing, Wang Min, Tian Zheng, Liu Xiang-rong, Lin Dong, Wang Jian-xiang
Institute of Hematology and Blood Diseases Hospital, CAMS and PUMC, Tianjin 300020, China.
Zhonghua Xue Ye Xue Za Zhi. 2005 Dec;26(12):728-31.
To explore the effect of 17-allylamide-17-demethoxygeldanamycin (17AAG), a heat shock protein 90 (HSP90) inhibitor, on the growth, differentiation and apoptosis of leukemic Kasumi-1 cells.
Kasumi-1 cells were treated with 17AAG at different concentrations in suspension culture. Cell proliferation was analysed by MTT assay, expression of myeloid-specific differentiation antigen and cell cycle by flow cytometry, cell apoptosis by annexin V staining, agarose gel electrophoresis and flow cytometry. KIT protein was analysed by Western blot and c-kit mRNA by RT-PCR.
17AAG treatment caused a dose-dependent inhibition of the cell proliferation with the IC(50) of 0.62 micromol/L. A dose-dependent increase in early apoptosis occurred at 24 hours treatment and in late apoptosis at 48 hours treatment. 17AAG induced a time- and dose-dependent increase in expression of myeloid cell surface protein CD11b and CD15, a progressive decline in S-phase cell fraction and an increase in G(0)/G(1) cells. When Kasumi-1 cells were incubated with 1 micromol/L of 17AAG, KIT protein began to decrease at 2 hours and KIT protein could hardly be detected at 20 hours, but c-kit mRNA was not decreased.
17AAG treatment of Kasumi-1 cells could lower KIT protein expression, inhibit cell proliferation, induce cell partial differentiation, apoptosis and accumulation in G(0)/G(1) phase.
探讨热休克蛋白90（HSP90）抑制剂17-烯丙基胺-17-去甲氧基格尔德霉素（17AAG）对白血病Kasumi-1细胞生长、分化及凋亡的影响。
在悬浮培养中用不同浓度的17AAG处理Kasumi-1细胞。通过MTT法分析细胞增殖，用流式细胞术分析髓系特异性分化抗原的表达及细胞周期，用膜联蛋白V染色、琼脂糖凝胶电泳和流式细胞术分析细胞凋亡。通过蛋白质印迹法分析KIT蛋白，通过逆转录-聚合酶链反应分析c-kit mRNA。
17AAG处理导致细胞增殖呈剂量依赖性抑制，半数抑制浓度（IC50）为0.62微摩尔/升。处理24小时时早期凋亡呈剂量依赖性增加，处理48小时时晚期凋亡增加。17AAG诱导髓系细胞表面蛋白CD11b和CD15的表达呈时间和剂量依赖性增加，S期细胞比例逐渐下降，G0/G1期细胞增加。当Kasumi-1细胞与1微摩尔/升的17AAG孵育时，KIT蛋白在2小时开始减少，20小时时几乎检测不到KIT蛋白，但c-kit mRNA未减少。
用17AAG处理Kasumi-1细胞可降低KIT蛋白表达，抑制细胞增殖，诱导细胞部分分化、凋亡及在G0/G1期蓄积。
