He Q, Na X
Research Laboratory of Blood Physiology, Hunan Medical University, 410078, Changsha, People's Republic of China.
Leuk Res. 2001 Jun;25(6):455-61. doi: 10.1016/s0145-2126(00)00153-3.
To evaluate the effects of a novel 2-aminosteroid, 2-(4'-methyl-1'-piperazinyl)-3alpha-hydroxyl-5alpha-androstane-17-one (KH), on in vitro murine WEHI-3B leukemia cells, semisolid colony culture, MTT assay, morphological examination, NBT reduction, NSAE test and ACP assay were used to determine proliferation and differentiation. It was found that the growth of leukemia cells in colony and liquid cultures was inhibited by KH (10(-8)-10(-4) mol/l) after treatment for 7 days. The percentages of NBT and NSAE positive cells were 71.17 and 79.25%, respectively, after treatment with KH (10(-8)-10(-6) mol/l) for 5 days. The morphology of treated leukemia cells was identified to be macrophage-like and these cells acquired significant ACP activities. It was indicated that the ACP enzyme activities were increased as high as two and three times of the control, respectively, after treatment with 10(-8) or 10(-5) mol/l KH for 6 days. It was also indicated by DNA fragmentation in gel electrophoresis that WEHI-3B cells were induced toward apoptosis by KH (10(-8)-10(-4) mol/l) when checked at day 5. The c-myc mRNA expressions in WEHI-3B cells were decreased by 58.7% after treatment with KH (10(-8) mol/l) for 5 days. Therefore, it is first reported here that KH, a novel 2-aminosteroid, could suppress proliferation and induce differentiation of WEHI-3B leukemia cells. These differentiated cells were mature macrophage-like cells and showed characteristics of functional phagocytes acquired with acid phosphatase activity. The mechanisms underlying the above effects involved the apoptosis of WEHI-3B leukemia cells and the down-regulation of c-myc oncogene expression. It is also shown that the counts of immature granulocytes and monocytes were significantly decreased in both peripheral blood and bone marrow of BALB/c leukemia mice after KH was administrated per os for 7 consecutive days with four doses (5, 10, 15 or 20 mg/kg day), respectively. It is also observed that the enlarged spleens in leukemia mice were decreased when compared with the control.
为评估新型2-氨基甾体2-(4'-甲基-1'-哌嗪基)-3α-羟基-5α-雄甾烷-17-酮（KH）对体外培养的小鼠WEHI-3B白血病细胞的作用，采用半固体集落培养、MTT法、形态学检查、NBT还原试验、非特异性酯酶（NSAE）试验和酸性磷酸酶（ACP）测定来确定细胞的增殖和分化情况。结果发现，经KH（10(-8)-10(-4)mol/L）处理7天后，白血病细胞在集落培养和液体培养中的生长均受到抑制。用KH（10(-8)-10(-6)mol/L）处理5天后，NBT阳性细胞和NSAE阳性细胞的百分比分别为71.17%和79.25%。经处理的白血病细胞形态被鉴定为巨噬细胞样，且这些细胞获得了显著的ACP活性。结果表明，用10(-8)或10(-5)mol/L KH处理6天后，ACP酶活性分别增加至对照的两倍和三倍。凝胶电泳中的DNA片段化结果也表明，在第5天检测时，KH（10(-8)-10(-4)mol/L）可诱导WEHI-3B细胞发生凋亡。用KH（10(-8)mol/L）处理5天后，WEHI-3B细胞中的c-myc mRNA表达下降了58.7%。因此，本文首次报道新型2-氨基甾体KH可抑制WEHI-3B白血病细胞的增殖并诱导其分化。这些分化细胞为成熟的巨噬细胞样细胞，并表现出具有酸性磷酸酶活性的功能性吞噬细胞的特征。上述作用的机制涉及WEHI-3B白血病细胞的凋亡和c-myc癌基因表达的下调。此外还表明，连续7天经口给予BALB/c白血病小鼠四剂（分别为5、10、15或20mg/kg/天）KH后，外周血和骨髓中未成熟粒细胞和单核细胞的数量均显著减少。同时还观察到，与对照组相比，白血病小鼠肿大的脾脏体积减小。
