过表达热休克蛋白22抑制造血系统恶性肿瘤细胞生长的研究  

Overexpression of heat shock protein 22 inhibits the growth of hematopoietic malignant tumor cells

作　　者：崔雪莹[1] 马小彤[2] 王丽娜[2] 王楠[2] 任倩[2] 林永敏[2] 

机构地区：[1]首都医科大学附属北京同仁医院血液科,100730 [2]中国医学科学院北京协和医学院血液学研究所血液病医院实验血液学国家重点实验室

出　　处：《白血病．淋巴瘤》2013年第12期706-711,共6页Journal of Leukemia & Lymphoma

基　　金：国家自然科学基金（81270634）;国家自然科学基金（81070426）;天津市应用基础重点项目（11JCZDJC18000）;高等学校博士学科点专项科研基金（20111106110036）

摘　　要：目的 探讨过表达热休克蛋白22（HSP22）对造血系统恶性肿瘤细胞生长和凋亡的影响.方法 构建含HSP22或空载体的慢病毒表达载体,感染K562和Namalwa细胞;荧光显微镜及流式细胞术观察感染效率;反转录-聚合酶链反应（RT-PCR）和Western blot鉴定HSP22表达.采用细胞计数法测定细胞增殖,流式细胞术检测细胞周期,甲基纤维素半固体集落形成实验测定集落数,Annexin VPE／7-AAD流式细胞术检测细胞凋亡.在裸鼠皮下注射K562和Namalwa细胞进行成瘤实验以观察成瘤体积.结果 与转导空载体的对照细胞相比,HSP22过表达能抑制Namalwa和K562细胞集落形成,每孔平均集落数转导pCDH1-MCS1-EF1-copGFP空载体的K562细胞为108,转导pCDH1-MCS1-EF1-copGFPHSP22的K562细胞为72,两者差异有统计学意义（P=0.000 16）;转导pCDH1-MCS1-EF1-copGFP空载体的Namalwa细胞为125,转导pCDH1-MCS1-EF1-copGFP-HSP22的Namalwa细胞为80,两者差异有统计学意义（P=0.000 37）.HSP22还能抑制Namalwa细胞体外增殖（第5天,P=0.015;第6天,P=0.042;第7天,P=0.048）,抑制K562细胞体内增殖（第21天,P=0.022）.K562、Namalwa细胞HSP22转导组与对照组间G0～G1期、G2～M期及S期细胞所占百分比差异均无统计学意义（均P＞ 0.05）.结论 过表达HSP22能够抑制造血系统恶性肿瘤K562和Namalwa细胞的生长.Objective To investigate the effects of overexpression of heat shock protein 22（HSP22） in hematopoietic malignant tumor cell lines.Methods A lentiviral system was used to mediate transduction of HSP22 complementary DNA-containing expression vector or empty vector into K562 and Namalwa cells.The transduction effeciency was tested by fluorescence microscope scan and flow cytometry.Semi-quantitative RT-PCR and Western blot were used to identify the expression levels of HSP22 mRNA and protein.Growth curve analysis,cell cycle analysis,colony-forming assay,tumor growth in nude mice and apoptosis analysis were used to evaluate the role of HSP22 in K562 and Namalwa cells.Results Lentivector expression systemmediated delivery of HSP22 into K562 and Namalwa cells can inhibit colony forming of K562 and Namalwa cells,the average numbers of colonies per well were 108,72,125 and 80 for K562-V,K562-H,Namalwa-V and Namalwa-H respectively （P =0.000 16 and 0.000 37 for K562 and Namalwa respectively）.HSP22 transduction can also inhibit proliferation of Namalwa cells in vitro （P =0.015,0.042 and 0.048 for day 5,6 and 7 respectively） and K562 cells in vivo （P =0.022 for day 21）.No significant difference in cell cycle and apoptosis was found in K562 and Namalwa cells compared with controls （all P ＞ 0.05）.Conclusion Overexpression of HSP22 could inhibit the growth of hematopoietic malignant tumor cell lines K562 and Namalwa.

关 键 词：血液肿瘤 热休克蛋白22 K562细胞 Namalwa细胞 细胞凋亡 

分 类 号：R733[医药卫生—肿瘤]