PPAR-γ配体联合顺铂对人肺癌PLA-801D细胞增殖抑制及诱导凋亡作用  

Inhibitory effects of ligand of PPAR-γ combined with DDP activation on proliferation of cells and induction of apoptosis of human pulmonary carcinoma PLA-801D cells in vitro

作　　者：曹岩[1] 刘红[2] 林栋[3] 郑永晨[1] 刘永红[4] 

机构地区：[1]吉林大学第二医院中心实验室,吉林长春130041 [2]吉林大学口腔医院修复科,吉林长春130041 [3]吉林大学白求恩医学部,吉林长春130021 [4]吉林省肿瘤医院电诊室,吉林长春130012

出　　处：《吉林大学学报（医学版）》2008年第3期433-438,共6页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅基金资助课题(20030434)

摘　　要：目的:探讨15-脱氧前列腺素J2(15d-PGJ2)联合顺铂(DDP)对人肺癌PLA-801D细胞生长的影响及其诱导凋亡的机制。方法:取生长良好的人肺癌PLA-801D细胞株,接种于96孔培养板,培养24 h后分别加入不同浓度15d-PGJ2(0、5、10、20、40、80μg.L-1)(单纯使用15d-PGJ2各组)和联合加入15d-PGJ2与DDP(3 mg.L-1)(15d-PGJ2+DDP各组)(0μg.L-1为对照组),作用24 h后用倒置显微镜观察每组细胞形态学的变化。采用MTT比色法检测15d-PGJ2联合顺铂对人肺癌PLA-801D细胞增殖抑制作用;DPA法检测其活性;流式细胞术检测其诱导细胞凋亡及周期的变化。结果:当低浓度15d-PGJ2(5、10和20μg.L-1),作用于人肺癌PLA-801D细胞时,细胞生长抑制率、凋亡率、DNA片段化比率和caspase-3活性与对照组比较差异均无显著性(P>0.05),当浓度为40μg.L-1时,细胞生长抑制率、凋亡率、DNA片段化比率和caspase-3活性均高于对照组(P<0.01);当15d-PGJ2(10μg.L-1)和DDP(3 mg.L-1)联合应用时,即可使人肺癌PLA-801D细胞生长抑制率、细胞凋亡率、DNA片段化比率和caspase-3活性均较单独使用15d-PGJ2明显增高(P<0.01)。当15d-PGJ2(40μg.L-1)作用人肺癌PLA-801D细胞24 h后,G0/G1期细胞比例明显高于对照组(P<0.01),S和G2/M期细胞比例均明显低于对照组(P<0.01)。结论:单独使用15d-PGJ2在高浓度时可以诱导人肺癌PLA-801D细胞凋亡,15 d-PGJ2与DDP联合应用时,前者低浓度即可诱导人肺癌PLA-801D细胞凋亡作用增强。Objective To investigate the effects of 15-deoxy-△12, 14-prostaglandin J2 （15d-PGJ2） and DDP on the growth of human pulmonary carcinoma PLA-801D cells and the mechanisms of apoptosis. Methods The human pulmonary carcinoma PLA-801D cells were selected and added to each well of 96-well place and cultivated for 24 h.Then the cells were treated with different concentrations of 15d-PGJ2 （0, 5, 10, 20, 40 and 80 μg·L^-l） or 15d- PGJ2 combined with DDP （3 mg·L） for 24 h. 0 μg·L^-1 15d-PGJ2 group was control group. The morphological changes of cells were observed under inverted microscope. Microculture tetrazolium （MTT） dye was applied to detect the proliferation of the human pulmonary carcinoma PLA-801D cells treated with 15d-PGJ2 and DDP. Diphenylamine assay （DPA） was used to evaluate the activation. Flow cytometry assay （FCM） was used to detect the apoptosis proportion and the changes of cell cycle. Results When the human pulmonary carcinoma PLA-801D cells were treated with low-concentration 15d-PGJ2 alone （ 5, 10 and 20 μg·L^-l）, no significant difference was observed in the inhibitory rate of cell growth and the apoptotic indexes such as the apoptosis proportion, the percent of DNA fragmentation and the activity of caspase-3 compared with control group （P〈0.05）; when 15d-PGJ2 concentration was 40 μg·L^-1 , the inhibitory rate of cell growth the apoptosis proportion and the percent of DNA fragmentation were higher than those in control group （P〈0.0l）. While the cells were treated with 15d-PGJ2 （10 μg·L^-1）combinated with DDP （3 mg·L ^-1）, the inhibitory rate of cell growth, the apoptosis proportion, the percent of DNA fragmentation and the activity of caspase-3 were significantly higher compared with 15d-PGJz alone （P〈0. 01）. when the human pulmonary carcinoma PLA-801D cells were treaed with 15d-PGJ2 alone for 24 h, and the 15d-PGJ2 concentration was 40 μg·L^-1, the cell proportion in G0/G1 phase was significantly higher than that in control group,

关 键 词：过氧化物酶增殖激活物受体-γ 15-脱氧前列腺素 人肺癌PLA-801D细胞株 细胞凋亡 细胞周期 顺铂 

分 类 号：R734.2[医药卫生—肿瘤]