雷帕霉素靶蛋白抑制剂PP242对急性T淋巴细胞白血病Jurkat细胞株增殖的影响  

Effects of mammalian target of rapamycin inhibitor PP242 on proliferation of acute T-lymphoblastic leukemia Jurkat cell line

作　　者：白波[1] 马梁明[1] 鹿育晋[1] 赵汀梓[1] 吴海燕[1] 赵小娟[1] 

机构地区：[1]山西医学科学院山西大医院血液科,太原030032

出　　处：《中国医药》2015年第4期520-522,共3页China Medicine

基　　金：山西省卫生和计划生育委员会科研课题（2014003）

摘　　要：目的 探讨雷帕霉素靶蛋白抑制剂PP242对急性T淋巴细胞白血病Jurkat细胞株增殖的影响及可能机制.方法 用含有10％胎牛血清的RPMI 1640培养液培养Jurkat细胞,调整细胞浓度为1×105/ml,接种于96孔培养板,每孔 180μ1细胞悬液.观察组加入不同浓度PP242,每孔20μ1,使其终浓度为0.1、0.5、1.0、5.0、10.0μmol/L.对照组每孔加入含有0.01％二甲基亚砜的RPMI 1640培养基20μl.采用四甲基偶氮唑盐法检测经不同浓度PP242处理的Jurkat细胞的增殖情况,用流式细胞仪检测不同浓度PP242作用48 h后Jurkat细胞的细胞周期改变和细胞凋亡情况.结果 培养24、48、72 h后,对照组吸光度值分别为0.569±0.023、0.901±0.022、1.199±0.041;1μmol/L观察组细胞吸光度值分别为0.513±0.014、0.764±0.019、0.955 ±0.016;5 μmol/L观察组细胞吸光度值分别为0.438±0.019、0.661 ±0.025、0.840±0.034;10 μmol/L观察组细胞吸光度值分别为0.407 ±0.011、0.613±0.026、0.791±0.012;浓度为1.0、5.0、10.0μmol/L的PP242能明显抑制Jurkat细胞的增殖.与对照组比较,1.0、5.0、10.0 μmol/L的PP242作用48 h后G0/G1期的Jurkat细胞百分比增高[（63.2±0.7）％、（65.8±0.4）％、（68.2±0.7）％比（56.9±0.6）％],差异均有统计学意义（均P＜0.05）.浓度为1.0、5.0、10.0 μmol/L的PP242作用48 h后,Jurkat细胞凋亡率分别为（1.7±0.6）％、（1.8±0.7）％、（2.4±0.8）％,对照组Jurkat细胞凋亡率为（1.6±0.6）％,差异无统计学意义（P＞0.05）.结论 PP242能明显抑制体外培养的急性T淋巴细胞白血病Jurkat细胞株的增殖,其机制可能是使Jurkat细胞的细胞周期阻滞于G0/G1期.Objective To investigate the effects and possible mechanisms of mammalian target of rapamy- tin inhibitor PP242 on proliferation of acute T-lymphoblastic leukemia Jurkat cells. Methods Jurkat cells were cultured with Roswell Park Memorial Institute （RPMI） 1640 medimn containing 10% fetal calf serum（ FCS）, and were seeded at a density of 1 × 105/ml into 96-well plates with cell suspension of 180 μl. Jurkat ceils were divided into experimental group added with 20 μl PP242 of different concentrations （ final acid concentration： 0.1 , 0.5, 1.0, 5.0, 10.0 μmol/L） and control group added with 20 μl RPMI 1640 medium containing 0.01% dimethyl sulfoxide （DMSO）. Proliferation of Jurkat cells was analyzed by MTT assay. Flow cytonletry was used to analyze tile cell cyc＇le and cell apoptosis of Jurkat cells 48 hours after treatment. Results The values of absorbance were 0.569 ±0. 023, 0. 901 ± 0. 022, 1. 199 ± 0. 041 in control group. Compared with control group, 1.0, 5.0, 10.0 Ixmoi/L PP242 significantly inhibited the proliferation of Jurkat cells 24, 48 and 72 hours after treatment （24 h A： 0.513 ±0.014, 0.438 ±.019, 0.407 ±.011 vs 0.569 ±.023, 48 h A： 0.764 ±.019, 0.661 ±0.025, 0.613 ±0.026 vs 0.901 ±0.022, 72 h A： 0.955 ±0.016, 0. 840 ±.034, 0.791 ±.012 vs 1. 199 ±0. 041 ）. The percentage of G0/G1 stage Jurkat cells incubated with 1.0, 5.0, 10.0 p, mol/L PP242 48 hours after treatment increased compared with control cells [（63.2 ±0.7）%, （65.8 ±.4）%, （68.2 ±0.7）% vs （ 56.9 ±0.6） % ] （ P 〈 0.05 ）. The apoptotic rates of Jurkat cells incubated with 1.0, 5.0, 10.0 μmol/L PP242 48 hours atier treatment were （ 1.7 ± 0.6） % , （ 1.8 ±0.7 ） % , （ 2.4 ±0.8 ） % , respectively, with no statistical difference with cmltrol cells I （ 1.6 ±0.6） % ] （ P 〈 0.05 ）. Conclusion PP242 can inhibit the proliferation of Jurkat cells, the mechanism of which is related with G0/G1 phase arrest action.

关 键 词：白血病 T细胞 JURKAT细胞 细胞周期 

分 类 号：R733.71[医药卫生—肿瘤]