Wang Yan-Fang, Song Quan-Sheng, Zhang Ying-Mei, Ma Da-Long, Wang Ying, Ke Xiao-Yan
Department of Hematology, The Third Hospital, Peking University, Beijing 100191, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Apr;18(2):277-81.
This study was aimed to investigate the sensitizing effect of recombinant human PDCD5 (rhPDCD5) protein on chemotherapy of U937 cell line and its mechanism. The flow cytometry was performed to assess the changes of cell apoptosis and cell cycle influenced by rhPDCD5. Hochst 33258 staining was used to observe morphology of the apoptotic cells. The activity change of caspase-3 was detected to analyse the possible mechanisms of rhPDCD5-induced apoptosis. RT-PCR was performed to observe the expression level of drug-resistant genes. The results showed that the percentage of apoptotic cells and the activity of caspase-3 remarkably increased in U937 cells treated with rhPDCD5 combined with chemotherapeutic drug; the cell cycle arrest induced by anti-tumor drug was also enhanced when combined with rhPDCD5; meanwhile, the expression levels of drug-resistant genes were down-regulated in jointly treated U937 cells. It is concluded that the chemosensitizing mechanisms of rhPDCD5 are complex. rhPDCD5 may increase the cytotoxicity of anti-tumor drugs by promoting the caspase-3-related apoptosis, influencing cell cycle, decreasing the expression of drug-resistant genes and reversing drug-resistance.
本研究旨在探讨重组人程序性细胞死亡蛋白5(rhPDCD5)对U937细胞系化疗的增敏作用及其机制。采用流式细胞术评估rhPDCD5对细胞凋亡和细胞周期的影响。用Hochst 33258染色观察凋亡细胞的形态。检测caspase-3的活性变化以分析rhPDCD5诱导凋亡的可能机制。进行逆转录聚合酶链反应(RT-PCR)观察耐药基因的表达水平。结果显示,rhPDCD5联合化疗药物处理的U937细胞中,凋亡细胞百分比和caspase-3活性显著增加;与rhPDCD5联合时,抗肿瘤药物诱导的细胞周期阻滞也增强;同时,联合处理的U937细胞中耐药基因的表达水平下调。结论是,rhPDCD5的化疗增敏机制复杂。rhPDCD5可能通过促进caspase-3相关凋亡、影响细胞周期、降低耐药基因表达及逆转耐药性来增加抗肿瘤药物的细胞毒性。