Pim-1激酶抑制剂SMI-4a抑制U937细胞增殖并诱导凋亡  

Pim-1 kinase inhibitor SMI-4a inhibits proliferation and induces apoptosis in U937 cells

作　　者：范蕊芳[1] 邹丽媛[1] 郝秀兰[2] 卢佩梅[1] 曾军荣[1] 蔡东兰[1] 刘相富[3] 

机构地区：[1]中山大学附属第三医院预防保健科,广东广州510630 [2]中山大学附属第三医院产科,广东广州510630 [3]中山大学附属第三医院输血科,广东广州510630

出　　处：《中国病理生理杂志》2017年第9期1625-1630,共6页Chinese Journal of Pathophysiology

基　　金：广东省科技计划(No.2011B2080701008)

摘　　要：目的:研究丝/苏氨酸蛋白激酶Pim-1抑制剂SMI-4a对人类急性髓系白血病细胞株U937的生长抑制、促凋亡作用及其可能机制。方法:CCK-8法检测不同浓度SMI-4a作用不同时间对U937细胞的生长抑制率;Annexin V-PI及Hoechst 33342染色法检测SMI-4a作用前后细胞凋亡情况,集落形成实验检测SMI-4a对U937细胞集落形成能力的影响;Western blot法检测SMI-4a对U937细胞核及细胞浆内β-catenin表达变化及细胞内凋亡相关蛋白的表达变化;免疫荧光法检测β-catenin在细胞内的表达变化。结果:CCK-8结果显示SMI-4a可以抑制U937细胞的活力,并呈时间和剂量依赖性;Annexin V-PI及Hoechst 33342染色结果显示SMI-4a可以促进U937细胞凋亡;集落形成实验证实SMI-4a可以抑制U937细胞的集落形成能力;Western blot实验结果显示SMI-4a作用于U937细胞48 h后细胞浆内的β-catenin表达增加,细胞核内的β-catenin表达减少,细胞内促凋亡蛋白Bax和PARP表达增强,抑凋亡蛋白Bcl-2表达明显减弱;免疫荧光进一步验证了SMI-4a作用后的U937细胞核内的β-catenin表达量明显减少。结论:SMI-4a诱导U937细胞凋亡是通过上调促凋亡基因的表达、下调凋亡抑制基因的表达来实现的。AIM： To study the growth-inhibiting and proapoptotic effects of Pim-1 kinase inhibitor SMI-4a on human acute myeloid leukemia cell line U937. METHODS： The effect of SMI-4a on U937 cell viability was measured by CCK-8 assay. The apoptotic rate was assessed by flow cytometry with Annexin V-PI staining and by fluorescence microscopy with Hoechst 33342 staining. Methylcellulose was used to assess colony formation ability of the cells. The expression of β-catenin in the cell cytosol and nucleus was detected by Western blot,and the expression of apoptosis-related proteins in the U937 cells was also examined. Intracellular distribution of β-catenin was detected by the method of immunofluorescence.RESULTS： SMI-4a inhibited the viability of U937 cells. Annexin V-PI staining showed that SMI-4a induced apoptosis in dose-and time-dependent manners. Hoechst 33342 staining also verified the apoptosis. SMI-4a significantly inhibited the colony formation capacity of the U937 cells. The results of Western blot demonstrated that SMI-4a upregulated the expression of PARP and Bax,downregulated the expression of Bcl-2 and change the distribution of β-catenin in intracellular compartment. Immunofluorescence observation found that SMI-4a decreased the expression level of β-catenin in the U937 cells. CONCLUSION： SMI-4a induces U937 cell apoptosis through regulating the expression of apoptosis-related genes.

关 键 词：Pim-1激酶 SMI-4a 急性髓细胞白血病 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤]