嵌合型溶瘤腺病毒SG235体外对白血病细胞抑制作用的实验研究  

Inhibitive effects of chimeric oncolytic adenovirus SG235 on leukemia cells in vitro

作　　者：杨春梅[1] 刘辉[1] 杨秀娣[1] 金洁[1] 钱文斌[1] 

机构地区：[1]浙江大学医学院附属第一医院,浙江杭州310003

出　　处：《浙江大学学报（医学版）》2010年第3期226-230,共5页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金杰出青年团队项目(R2090392);"十一五"国家科技支撑计划(2008BAI61B01)

摘　　要：目的:研究嵌合型溶瘤腺病毒(CRAd)SG235的抗白血病作用。方法:用流式细胞术(FACS)检测原代白血病细胞CD46的表达和SG235对多种白血病细胞株的感染率,MTT法研究SG235对Kasumi-1细胞的生长抑制,Annexin-V/PI染色和TUNEL法观察SG235诱导白血病细胞凋亡的作用,Western blot检测CRAd对磷酸化Akt蛋白表达的影响。结果:大多数急性白血病患者原代白血病细胞表达CD46。用50病毒感染复数(MOI)的SG235-EGFP感染Mutz-1、Kasumi-1、K562、HL60、Molt-4、RPMI8226、L428、Jurkat细胞,感染率分别为45.1%、35.7%、54.2%、37.0%、30.1%、67.1%、17.2%、33.1%。SG235对Kasumi-1细胞有显著的生长抑制作用,作用24 h和48h IC50值分别为79.0 MOI、38.4 MOI。SG235能诱导多种白血病细胞株凋亡,而且呈剂量依赖性;并能抑制Kasumi-1细胞磷酸化Akt活性。结论:嵌合型溶瘤腺病毒SG235能有效地感染白血病细胞,并能抑制白血病细胞的生长,诱导其凋亡。Objective： To investigate the inhibitive effects of chimeric oncolytic adenovirus SG235 on leukemia cells in vitro.Methods： The ability of SG235 to infect leukemia cells and the expression of CD46 on blasts from the patient with leukemia were detected by flow cytometry（FACS）.The cytotoxicity of the virus was evaluated by MTT assay.Apoptosis induced by SG235 was detected with Annexin-V/PI staining and TUNEL assay followed by FACS analysis.The expression of pAkt in leukemia cells was determined by Western blot analysis.Results： The majority of leukemia cells from the patient with acute leukemia was CD46-positive.GFP-positive cells were 45.1%,35.7%,54.2%,37.0%,30.1%,67.1%,17.2%,and 33.1% in Mutz-1,Kasumi-1,K562,HL60,Molt-4,RPMI8226,L428,and Jurkat cell lines treated with SG235-EGFP vector at MOI（multiplicity of infection） of 50 for 48 h.SG235 treatment resulted in marked growth inhibition and apoptosis of Kassumi-1 cells,and also significantly inhibited expression of p-Akt.Conclusion： The chimeric oncolytic adenovirus SG235 can infect leukemia cell effectively and results in the growth inhibition and apoptosis of Kasumi-1 cells in vitro.

关 键 词：白血病/治疗 白血病/病理学 基因疗法 腺病毒科/遗传学 肿瘤细胞 培养的 细胞凋亡/药物作用 

分 类 号：R733.7[医药卫生—肿瘤]