去甲斑蟊素诱导T淋巴细胞白血病Jurkat细胞凋亡作用及其机制  

Apoptosis and mechanism study on jurkat T lymphoblastic leukemia cells induced by Norcantharidin(NCTD)

作　　者：李晓辉[1] 王亚君[1] 陈丽丽[1] 宋飞飞[1] 张剑白[1] 

机构地区：[1]哈尔滨医科大学附属第一医院儿科,哈尔滨150001

出　　处：《中国小儿血液与肿瘤杂志》2016年第2期81-86,共6页Journal of China Pediatric Blood and Cancer

基　　金：哈尔滨市科技创新人才研究专项资金优秀学科带头人(2012RFXXS017)

摘　　要：目的研究去甲斑蝥素(norcantharidin,NCTD)在体外对急性T淋巴细胞白血病细胞株Jurkat细胞凋亡的作用及其机制。方法取对数生长期的Jurkat细胞分别加入不同浓度的NCTD和(或)10 mg/L的长春新碱(VCR),倒置显微镜下观察不同时间Jurkat细胞形态、密度及结构的变化;采用MTT比色法测定NCTD及VCR两种药物作用于细胞72 h后对细胞增殖抑制作用的影响;实时荧光定量RT-PCR法对p53及survivin基因表达量进行检测。结果 NCTD及VCR作用组细胞形态出现不规则,胀大、变形现象明显,分布稀疏,大量细胞死亡,两者在相同时间点上对细胞的影响无明显差别。MTT法检测结果表明:NCTD增殖抑制作用呈现时间-浓度依赖关系,最佳浓度及时间在20 mg/L,作用72 h时。NCTD与VCR两者对Jurkat细胞均有抑制作用(P<0.05),两者的抑制率相比无显著性差异(P>0.05),且两者联合可增加细胞增殖抑制率(P<0.05)。应用实时荧光定量RT-PCR技术定量表达p53及survivin基因,结果显示NCTD组p53基因表达较对照组细胞株明显上升(P<0.001),相反survivin基因较对照组明显下降(P<0.01)。结论(1)NCTD抑制Jurkat细胞株的增殖抑制作用具有时间-浓度依赖关系;(2)NCTD与VCR对细胞增殖均有抑制作用,两者的影响无显著性差异,且两者联合可增强对细胞的增殖抑制作用;(3)NCTD促进Jurkat细胞株细胞凋亡、抑制增殖的机制可能与上调p53基因表达量及下调survivin基因表达量有关。Objective To study the effect of Norcantharidin （NCTD） on cell apoptosis of Jurkat cells, an acute T cell leukemia cell line, and the possible mechanisms including the expression of apoptosis related genes. Methods Morphology, density and structural changes of Jurkat cells in logarithmic were observed under an inverted microscope after treatment with NCTD at different concentrations or VCR at 10mg/L. Using MTF method, the effects of different concentrations of both NCTD and VCR treatment on cell proliferation were measured after 72h of treatment. The expression level of p53 and survivin were determined using real-time fluorescent quantitative PCR （RT-PCR）, to further explore the mechanism of NCTD induction of cell apoptosis. Results （ 1 ） Cells treated by NCTD and VCR showed swollen and deformed morphology, which were sparsely distributed with high rate of celldeath. Effects of the two drugs were not significantly different. （2） Inhibition of proliferation by NCTD was in time-dependent manner. The optimal NCTD concentration and treatment time were 20mg/L and 72 hours, respectively. MTT test showed that NCTD and VCR inhibited the proliferation of Jurkat cells and decreased the cell survival rate （P 〈 0.05 ）. Cells treated by NCTD and VCR did not show significant difference （ P 〉 0.05 ）. Treatment by NCTD and VCR in combination increased the cell proliferation decreased compared with the NCTD depends on time and control group （P 〈0.01 ）. drug concentration. Both Conclusions Inhibition of cell proliferation by NCTD and VCR inhibit cell proliferation, their effects are not significantly different, and the combination of the two drugs increases the inhibition effect. The mechanism of NCTD-induced apoptosis and cell proliferation inhibition could be related with the up-regulation of p53 expression and down-regulation of survivin expression.

关 键 词：T淋巴细胞白血病 JURKAT细胞株 去甲斑蝥素 p53基因 SURVIVIN基因 

分 类 号：R733.7[医药卫生—肿瘤]