曲古抑菌素A对外周血自然杀伤细胞活性和功能的影响

山东大学学报(医学版) ›› 2009, Vol. 47 ›› Issue (8): 50-54.

曲古抑菌素A对外周血自然杀伤细胞活性和功能的影响

王向玲¹，纪春岩¹，Magnus Nordenskjld ²，JanInge Henter ³，郑成云^2,3,4

1. 山东大学齐鲁医院血研室，济南 250012； 2. 瑞典卡罗林斯卡大学医院临床分子医学中心，斯德哥尔摩 171 76；
3. 瑞典卡罗林斯卡大学医院小儿癌症研究中心，斯德哥尔摩 171 76； 4. 山东大学第二医院血液科，济南 250033

收稿日期:2008-11-11 发布日期:2009-08-16
通讯作者: 郑成云（1962- ），男，博士，主要从事恶性血液病的基础与临床研究。 Email: chengyun.zheng@ki.se
作者简介:王向玲(1971- )，女，博士，主要从事恶性血液病的基础与临床研究。
基金资助:
国家自然科学基金资助项目（No.30670903），瑞典研究委员会及瑞典癌症基金会资助项目。

Effects of trichostatin A effects on natural killer cell survival and activity

WANG Xiangling ¹, JI Chunyan ¹, Nordenskjld Magnus ², Henter JanInge ³, ZHENG Chengyun ^2,3,4

1. Department of Hematology, Qilu Hospital of Shandong University, Jinan 250012, China;
2. Clinical Genetics Unit, Department of Molecular Medicine and Surgery, Karolinska University Hospital, Karolinska Institutet,
Stockholm 171 76, Sweden; 3. Childhood Cancer Research Unit, Department of Woman and Child Health,
Karolinska University Hospital, Karolinska Institutet, Stockholm 171 76, Sweden;
4. Department of Hematology, The Second Hospital of Shandong University, Jinan 250033, China

Received:2008-11-11 Published:2009-08-16

摘要/Abstract

摘要：

目的探讨曲古抑菌素A（TSA）对正常人外周血自然杀伤细胞(NK)活性和功能的影响。方法从正常人外周血单个核细胞（PBMCs）分离淋巴细胞（PBLs）（主要包括T细胞和NK细胞）或纯化NK细胞。体外应用白介素2（IL2）诱导PBLs 72?h产生淋巴因子激活的杀伤（LAK）细胞。将PBLs, LAK或NK细胞进行体外培养，给予不同浓度的TSA处理12、24、48?h。流式细胞术检测NK细胞和T细胞的凋亡；采用51Cr释放试验检测NK细胞的杀伤功能。结果TSA以剂量和时间依赖方式诱导PBLs和LAK细胞凋亡，TSA处理组淋巴细胞及LAK细胞凋亡百分率明显高于对照组（P＜0.05）；TSA同样可诱导静止期NK细胞的凋亡，0.1?μmol/L TSA作用于NK细胞24?h后，36%NK细胞凋亡，显著高于未处理组（P＜0.05）；0.1?μmol/L TSA作用于NK细胞１2?h后，其杀伤白血病细胞的能力明显低于对照组（P＜0.01）。结论TSA能够诱导人外周血静止期NK细胞及LAK细胞发生凋亡，并影响NK细胞的细胞毒作用。

关键词:
组蛋白去乙酰化酶抑制剂；自然杀伤细胞；凋亡；细胞毒作用

Abstract:

To explore the potential regulatory effects of trichostatin A (TSA) on survival and activity of natural killer (NK) cells. MethodsHuman peripheral blood lymphocytes（PBLs）or NK cells were separated or isolated from peripheral blood mononuclear cells (PBMCs) of healthy blood donors. Lymphokine activated killer (LAK) cells were generated by culturing PBLs in the presence of interleukin2 for 72?h. The cells were cultured in the presence or absence of TSA at the concentrations（0.01，0.1 and/or 1?μmol/L）for 12?h, 24?h and/or 48?h, respectively. Apoptotic cells in PBLs, LAK or purified NK cells were cytometrically quantified by dual labeling of propidium iodide (PI) and AnnexinV (AV). The proportion of apoptotic cells in PBLs were further defined by combined staining of AV, antiCD56 and antiCD3 antibodies, and flow cytometry method (FCM). NK cell cytolytic activity against leukemia cells (K562 leukemic cell line) was evaluated using a standard chromium51 release assay. ResultsTSA markedly induced apoptosis in PBLs and LAK cells in a dose and timedependent manner. The percentages of total apoptotic cells (AV+PI plus AV+PI+) in the TSA treatedPBLs or LAK cells were significantly higher than that in the untreated ones（P＜0.05）. Moreover, the percentage of total apoptotic cells in the purified resting NK cells treated by TSA(0.1?μmol/L) for 24?h was significantly higher than that in the control NK cells (P＜0.05). Importantly, the cytolytic activity of resting NK cells against leukemia cells significantly decreased after treatment with 0.1?μmol/L TSA for 12?h, as compared to the control NK cells（P＜0.01）. ConclusionsThe histone deacetylase inhibitor TSA can induce apoptosis in NK cells and LAK cells, and can affect NK cell cytotoxicity.

Key words: Histone deacetylase inhibitor; Natural killer cell; Apoptosis; Cytotoxicity

中图分类号:

R331.144

. 曲古抑菌素A对外周血自然杀伤细胞活性和功能的影响[J]. 山东大学学报(医学版), 2009, 47(8): 50-54.

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