YC-1抑制人类白血病细胞株U937、THP-1增殖作用的研究  

Research on inhibitory effect of YC-1 on proliferation of human leukemia U937,THP-1 cell lines

作　　者：张颖[1] 李蔚 邱林 程焕臣 赵雪飞 马军 赵瑛[1] 

机构地区：[1]哈尔滨商业大学药学院,150076 [2]哈尔滨市血液病肿瘤研究所

出　　处：《白血病．淋巴瘤》2012年第8期449-452,共4页Journal of Leukemia & Lymphoma

基　　金：基金项目：黑龙江省卫生厅科技计划（2011-520）;中国博士后科学基金（20110491102）

摘　　要：目的探讨YC-1对人类白血病细胞株U937和THP-1增殖、凋亡、周期及Caspase-3、-8、-9蛋白表达的影响。方法实验分为YC-1组（1．0、3．0、10．0umol／L），以不加YC-1组为对照。四甲基偶氮唑蓝比色（MTT）法检测细胞增殖；流式细胞术（FCM）检测细胞凋亡及周期；Western blot检测Caspase-3、-8、-9蛋白表达的变化。结果1．0、3．0、10．0umol／LYC-1以剂量依赖方式抑制U937、THP-1细胞增殖，24h细胞存活率分别为（76．5±4．4）％、（68．7±6．8）％、（60．9±13．2）％，（94．1±1．4）％、（81．4±2．0）％、（72．7±3．0）％，与对照组的100％比较差异均有统计学意义（F=15．870、126．629，均P〈0．01）。U937、THP-1细胞经1．0、3．0、10．0umol／L YC-1作用24h后，凋亡率分别为（40．7±1．0）％、（55．6±2．3）％、（71．8±1．5）％，（54．6±2．0）％、（50.3±3．5）％、（59．6±4．6）％，与对照组的（4．7±1．4）％、（1．8±1．0）％比较差异均有统计学意义（F=937．229、200．447，均P〈0．01），但细胞周期无明显变化。经1．0、3．0、10．0umol／L YC-1作用24h后，U937细胞Cleaved Caspase-3、-8蛋白表达上调，Caspase-9无变化，THP-1细胞Cleaved Caspase．3蛋白表达上调，Caspase-8、-9无变化。结论YC-1能够抑制U937、THP-1细胞增殖，诱导细胞凋亡，但对周期无影响，其诱导白血病细胞凋亡的机制可能与Caspase蛋白活化有关。Objective To delineate the potency of YC-1 on the proliferation, apoptosis, cell cycle and the protein expression of Caspase-3, -8, -9 in U937 and THP-1 leukemia cell lines. Methods MTT assay was performed to detect proliferation. Flow cytometry was used to measure the apoptosis and cell cycle. The expression of Caspase-3, -8 and -9 were detected by Western blot. Results The MTT assay showed that cell proliferation was inhibited in a concentration-dependent manner in 1.0, 3.0, 10.0 umol/L YC-l-treated U937 and THP-1 cells. The survival rates for YC-1 after 24 h in U937 cells were （76.5±4.4） %, （68.7±6.8） %, （60.9±13.2） % respectively and （94.1±1.4） %, （81.4±2.0） %, （72.7±3.0） % respectively in THP-1 cell, compared with the control group （100 %）, there were significant differences （F = 15.870, 126.629, P 〈 0.01）. The apoptosis rates for 1.0, 3.0, 10.0 umol/L YC-1 after 24 h were （40.7±1.0） %, （55.6±2.3） %, （71.8±1.5） % respectively in U937 cells and （34.6±2.0） %, （50.3±3.5） %, （59.6±4.6） % respectively in THP-1 cells. With the control group （4.7±1.4） %, （1.8±1.0） %, there were significant difference （F = 937.229, 200.447, P 〈 0.01）. However, there was no significant difference for cell cycle. In addition, Cleaved Caspase-8 and Cleaved Caspase-3 expression after 1.0, 3.0, 10.0 umol/L YC-1 treated for 24 h were significantly higher than control, but the expression of Caspase-9 did not appear significant change in U937 cells. As the same concentration and time point, Cleaved Caspase-3 expression increased with no change of Caspase-9 or Caspase-8 in THP-1 cells. Conclusion YC-1 effectively suppress the proliferation with little effect on cell cycle, but induce the apoptosis, have no effect on cell cycle, and the mechanism of apoptosis may be related to the Caspase activation in U937 and THP-1 cell lines.

关 键 词：白血病 实验性 缺氧诱导因子-1抑制剂 细胞周期 细胞凋亡 半胱氨酸天冬 氨酸蛋白酶 

分 类 号：R733.7[医药卫生—肿瘤]