组蛋白去乙酰化酶抑制剂提高腺病毒对K562细胞转染及杀伤效率的研究  

Enhancive effect of histone deacetylase inhibitor trichostatin a on transfection and antitumor efficiency of adenovirus in human chronic myelogenous leukemia cell line K562

作　　者：曹阳[1] 黄晓园[1] 庄亮[1] 李伟[1] 周剑峰[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院血液内科,武汉430030

出　　处：《白血病．淋巴瘤》2007年第6期401-404,共4页Journal of Leukemia & Lymphoma

基　　金：国家自然科学基金资助(30371657)

摘　　要：目的研究通过观察组蛋白去乙酰化酶(HDAC)抑制剂曲古抑菌素 A(TSA)对人类白血病细胞系 K562柯萨奇-腺病毒受体(CAR)表达水平的影响,并探讨 HDAC 抑制剂在以腺病毒为载体的基因治疗中的应用价值。方法在 mRNA 和蛋白水平检测 TSA 处理 K562细胞前后 CAR 的表达情况,采用流式细胞术测定病毒的转染效率,四甲基偶氮唑蓝(MTT)法检测腺病毒及其携带的基因的体外抗瘤效应。结果 TSA 处理后,K562细胞 CAR 的 mRNA 和蛋白表达明显增高;流式细胞仪分析ADV-GFP 转染 K562后 GFP 的表达发现:未处理组的转染率为(8.74±0.34)%,1、10和100 nmol/LTSA 处理细胞48h 后的转染率分别为(12.26±0.55)%、(20.83±1.22)%和(28.66±0.43)%。未处理组与处理组间及各处理组间转染效率的差异有统计学意义(P<0.05)。MTT 结果表明腺病毒 M1介导的体外杀伤效应 TSA 处理组比未处理组增强(P<0.05),TSA 各处理组间的体外杀伤效应随 TSA 浓度的提高而增强(P<0.05)。结论低浓度的 TSA 可以增强腺病毒对 K562细胞的感染及杀伤效率,可以提高以腺病毒为载体的血液肿瘤基因治疗的疗效。Objective To observe the influence of trichostatin A(TSA)a histonedeacetylase(HDAC) inhibitor,to CAR on the transfection efficiency of adenovirus in human chronic myelogenous leukemia cell line K562,and explore the possible application to adenovirus based gene therapy.Methods mRNA and pro- tein levels of CAR on K562 cells were detected by semiquantative reverse transcription-polymerase chain re- action(RT-PCR)and Western blot before and after treatment of TSA.Transfcction efficiency of adenovirus was valued by flow eytometry(FCM).In vitro antitumor effect of M1 were detected by MTT assay.Results After treatment of TSA,mRNA and protein levels of CAR on K562 cells were obviously increased.transfection rates of adenovirus were(8.74±0.34)% in untreated group,(12.26±0.55)% in 1 nmol/L of TSA treated group, (20.83±1.22)% in 10 nmol/L of TSA treated group and(28.66±0.43)% in 100 nmol/L of TSA treated group. There were significant differences between untreated group and TSA treated group(P<0.05).The TSA treated groups also showed statistical differences(P<0.05).In vitro antitumor effect of M1 was ehaneive in TSA(1, 10,100 nmol/L)treated groups compared with that in untreated group(P<0.05).The statistical differences in TSA treated groups were also significant(P<0.05).Conclusion TSA could enhance transfection and antitu- mot efficiency of adenovirus in K562 cells,and may be useful in adenovirus based gene therapy for leukemia treatment.

关 键 词：K562细胞 组蛋白脱乙酰基酶类 TRICHOSTATIN A 腺病毒 转染 杀伤效率 

分 类 号：R733.7[医药卫生—肿瘤]