Cai Zhen, Bao Han-ying, Ludwig Wolf-Dieter, Wuchter Christian
The First Affiliated Hospital, Medical College of Zhejiang University, Hangzhou 310003, China.
Zhonghua Xue Ye Xue Za Zhi. 2004 Jan;25(1):26-30.
To investigate the expression of apoptotic protein inhibitors, survivin and XIAP, in patients with myelodysplastic syndromes (MDS) and in the cell line MUTZ-1, as well as to explore the possible mechanisms of homoharringtonine (HHT) in the treatment of MDS.
Bone marrow samples from 47 patients with de novo MDS at diagnosis were examined and bone marrow samples from 15 normal donors were used as control. A MDS-RAEB cell line MUTZ-1 was used as in vitro model. Detection of apoptotic cells and cell cycle analysis were performed with flow cytometry (FACS). The expression of apoptotic protein inhibitor survivin and XIAP in the MDS cells were detected by RT-PCR technique. MUTZ-1 were treated with antisense oligodeoxynucleotide (AS-ODNs) of survivin and or HHT, the effects were evaluated by cell viability and cell apoptosis.
Survivin mRNA positive rate in MDS were significantly higher than that in normal controls (38.3% and 0, respectively, P < 0.01), and the positive rate in high risk group (RAEB, RAEBT and CMML) was significantly higher than that in RA/RAS group (53.6% and 16.7%, respectively, P < 0.05). XIAP was expressed in all untreated MDS and healthy controls. XIAP mRNA expression in high risk group was significantly higher than that in RA/RAS subtypes and healthy controls (1.55 +/- 0.34, 0.74 +/- 0.24, and 1.01 +/- 0.28, respectively, P < 0.01). However, XIAP mRNA expression was significantly lower in RA/RAS subtypes than in healthy control (0.74 +/- 0.24 and 1.01 +/- 0.28, P < 0.054). Apoptosis peak detected by FACS analysis and positive Annexin V FITC staining on cell membrane indicated that HHT could induce MUTZ-1 cell undergoing apoptosis in dose- and time-dependent manners. Treatment of MUTZ-1 cells with HHT revealed that HHT could significantly down-regulate survivinexpression but had no significant effect on XIAP expression in the cells. AS-ODNs of survivin could inhibit MUTZ-1 cells growth, induce them to apoptosis and sensitize them to HHT.
The expression levels of survivin; Institute of Hematology, Oncology and Tumor Immunology, Robert Roessle Clinic, Humboldt University, Berlin, Germany (Wolf Dieter Ludwig, Christian Wuchter) and XIAP vary in different subtypes of MDS patients, suggesting that the proteins may play an important role in the pathogenesis of the disease. Down-regulation of survivin in MUTZ-1 cells may be one of the mechanisms that HHT induces apoptosis of MDS cells.
研究凋亡蛋白抑制因子survivin和XIAP在骨髓增生异常综合征（MDS）患者及细胞系MUTZ-1中的表达情况，并探讨高三尖杉酯碱（HHT）治疗MDS的可能机制。
检测47例初诊的原发性MDS患者的骨髓样本，以15例正常供者的骨髓样本作为对照。采用MDS-RAEB细胞系MUTZ-1作为体外模型。运用流式细胞术（FACS）进行凋亡细胞检测和细胞周期分析。通过RT-PCR技术检测MDS细胞中凋亡蛋白抑制因子survivin和XIAP的表达。用survivin反义寡脱氧核苷酸（AS-ODNs）和/或HHT处理MUTZ-1细胞，通过细胞活力和细胞凋亡评估其效果。
MDS患者中survivin mRNA阳性率显著高于正常对照组（分别为38.3%和0，P<0.01），高危组（RAEB、RAEBT和CMML）的阳性率显著高于RA/RAS组（分别为53.6%和16.7%，P<0.05）。所有未经治疗的MDS患者和健康对照中均有XIAP表达。高危组XIAP mRNA表达显著高于RA/RAS亚型和健康对照（分别为1.55±0.34、0.74±0.24和1.01±0.28，P<0.01）。然而，RA/RAS亚型中XIAP mRNA表达显著低于健康对照（0.74±0.24和1.01±0.28，P<0.054）。FACS分析检测到的凋亡峰以及细胞膜上Annexin V FITC阳性染色表明，HHT能以剂量和时间依赖的方式诱导MUTZ-1细胞凋亡。用HHT处理MUTZ-1细胞显示，HHT能显著下调survivin表达，但对细胞中XIAP表达无显著影响。survivin的AS-ODNs能抑制MUTZ-1细胞生长，诱导其凋亡并使其对HHT敏感。
survivin和XIAP在不同亚型的MDS患者中的表达水平不同，提示这些蛋白可能在疾病发病机制中起重要作用。MUTZ-1细胞中survivin的下调可能是HHT诱导MDS细胞凋亡的机制之一。 德国柏林洪堡大学罗伯特·罗斯勒诊所血液学、肿瘤学和肿瘤免疫学研究所（沃尔夫·迪特尔·路德维希、克里斯蒂安·武赫特）
