蟾蜍灵在体内外抑制急性红白血病细胞K562增殖及下调WT1表达的研究  

Bufalin inhibits proliferation and downregulates expression of WT1 in K562 cells in vivo and vitro

作　　者：汪丽佩[1] 李天一[1] 高瑞兰[2] 杜月光[1] 赵燕娜[2] 

机构地区：[1]浙江中医药大学基础医学院病理学教研室,浙江杭州310053 [2]浙江中医药大学附属第一医院血液病研究所,浙江杭州310006

出　　处：《中国药理学通报》2016年第2期229-233,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81403223);浙江省自然科学基金项目(No LQ14H290003);浙江省医药卫生科技计划项目(No 2014KYA151)

摘　　要：目的研究蟾蜍灵在体内外抗K562细胞增殖及对WT1表达影响。方法半固体集落形成实验检测蟾蜍灵对K562细胞的增殖抑制作用,流式细胞术分析蟾蜍灵对K562细胞周期的影响,Western blot观察蟾蜍灵对WT1表达作用。通过高致瘤性K562细胞制备裸鼠皮下荷瘤模型,分组为模型组、蟾蜍灵组及高三尖杉酯碱组,比较各组皮下瘤体积与重量、病理形态学改变及WT1蛋白表达情况。结果①半固体集落形成实验、流式及Westem blot结果显示,蟾蜍灵能明显抑制K562细胞增殖,阻滞细胞在G_0/G_1期,并下调其WT1蛋白表达,呈剂量依赖性;②蟾蜍灵组和阳性对照组裸鼠肿瘤的抑瘤率均明显高于模型组(P<0.05),且蟾蜍灵组和阳性对照组裸鼠皮下瘤重量均明显低于模型组(P<0.05);③病理切片显示,蟾蜍灵导致K562皮下瘤组织内肿瘤细胞大量坏死、凋亡,继发出血及纤维化等改变;并明显抑制K562皮下瘤组织中WT1蛋白表达水平。结论蟾蜍灵在体外可抑制K562细胞增殖及阻滞细胞周期于G_0/G_1期并下调其WT1蛋白表达,在体内可明显抑制裸鼠K562皮下瘤体积和重量,导致皮下瘤组织坏死、凋亡,并可下调其WT1蛋白的表达。Aim To investigate the effect of bufalin on proliferation and expression of WT1 in K562 cells.Methods The colony number of K562 cell was detected with semi-solid culture assay.The cell cycle was measured by flowcytometry,and the expression of WT1 was observed with immunocytochemistry.Subcutaneous tumor models established by K562 cells in BALB/C nu/nu mice were divided into three groups,including model group,bufalin group and positive control group.After 21 days,the subcutaneous tumors were removed for calculating the inhibitory rate of tumor growth.HE staining and immunohistochemistry were used to observe the morphological changes and the expression of WT1.Results ① Bufalin could significantly decrease the colony number of K562 cell,arrest it at G_0/G_1phase and down-regulate its expression of WT1 in a dose-dependent manner.② Compared with the model group,the tumor inhibitory rate was much higher,while the volume and the weight were obviously lower in the other two groups.③ Bufalin could induce apoptosis,necrosis,hemorrhage and fibrosis with HE staining,and down-regulate the expression of WT1.Conclusion Bufalin could inhibit the proliferation,arrest the cell cycle at G_0/G_1 phase and down-regulate the expression of WT1 in vitro.Bufalin could inhibit the tumor inhibitory rate,the volume and the weight of the subcutaneous tumors,induce apoptosis,necrosis,hemorrhage and fibrosis with HE staining and downregulate the expression of WT1.

关 键 词：蟾蜍灵 WT1 K562 细胞周期 G0/G1期 皮下瘤 

分 类 号：R-332[医药卫生] R282.74