Synergistic antitumoral efficacy of a novel replicative adenovirus SG611-PDCD5 and daunorubicin in human leukemic cells.
Synergistic antitumoral efficacy of a novel replicative adenovirus SG611-PDCD5 and daunorubicin in human leukemic cells.
复制标题
新型复制腺病毒SG611-PDCD5和柔红霉素在人白血病细胞中的协同抗肿瘤功效
DOI：
10.2147/ott.s167868
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发表时间：
2018
影响因子：
4
通讯作者：
Ruan GR
中科院分区：
文献类型：
--
作者：
Zhou YL;Yao QM;Zhou J;Chang Y;Li JL;Wang YZ;Wu HP;Chen YH;Liu YR;Huang XJ;Ruan GR
柔红霉素是一种传统的化疗药物，在白血病治疗中起着关键作用。然而，剂量相关的毒性仍然是一个相当大的挑战。PDCD 5基因在白血病等多种肿瘤中表达下调，可能为白血病的诊断和治疗提供潜在的靶点。本研究旨在构建携带PDCD 5基因表达盒的三重调控溶瘤腺病毒（SG 611-PDCD 5），并探讨SG 611-PDCD 5联合小剂量柔红霉素对白血病细胞的联合抗肿瘤作用。材料与方法根据提供者的说明培养多种白血病细胞系，包括K562、MEG-01、KG-1a、HL-60、SUP-B15和BV-173。重组质粒经酶切和PCR鉴定，证实了质粒的插入和定向。构建的条件复制型SG 611-PDCD 5的肿瘤选择性复制及其与柔红霉素组合的抗肿瘤功效在体外白血病细胞系和裸鼠异种移植模型中进行了表征。使用细胞计数试剂盒-8检测细胞活力。使用流式细胞术和石蜡包埋的肿瘤组织中使用末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）测定在整个活细胞中检测细胞凋亡。结果成功构建了携带SG 611-PDCD 5的三重调控CRAd的K562细胞和裸鼠移植瘤模型。SG 611-PDCD 5与低剂量柔红霉素联合体外治疗以剂量依赖性方式在白血病细胞中引起更有效的抗增殖和促凋亡作用。Chou-Talalay分析揭示了在所有上述细胞系中的协同抗增殖作用。在裸鼠异种移植模型中，对照组、柔红霉素组、SG 611-PDCD 5组和联合治疗组在第10天的肿瘤大小分别为170.1±47.8、111.9±81.1、60.7±12.3和33.2±17.5 mm 3（均P<0.05）。TUNEL检测结果显示，SG 611-PDCD 5+柔红霉素组的凋亡细胞数明显多于SG 611-PDCD 5或柔红霉素组（分别为25±0.82、12.5±2.27和7.8±2.67个凋亡细胞/视野）（P<0.05）。结论SG 611-PDCD 5联合柔红霉素治疗白血病，可提高柔红霉素的化疗敏感性，降低其剂量相关毒性。
Background Daunorubicin is a traditional chemotherapeutic agent that plays a pivotal role in leukemia therapy. However, the dose-related toxicity remains a considerable challenge. The apoptosis-regulating gene, PDCD5, is downregulated in various tumors, including leukemias, and may provide a potential target for the diagnosis and treatment of leukemia. The purpose of this study was to construct a triple-regulated oncolytic adenovirus carrying a PDCD5 gene expression cassette (SG611-PDCD5) and explore the combined antitumor efficacy of SG611-PDCD5 in combination with low dose daunorubicin on leukemic cells. Materials and methods A variety of leukemic cell lines, including K562, MEG-01, KG-1a, HL-60, SUP-B15, and BV-173, were cultured according to the providers’ instructions. The insertion and orientation of all recombined plasmids were confirmed by restriction enzyme digestion and PCR. The tumor-selective replication of the constructed conditionally replicating SG611-PDCD5 and its antitumor efficacy in combination with daunorubicin were characterized in leukemic cell lines in vitro and in a nude mouse xenograft model. Cell viability was detected using cell-counting kit-8. Apoptosis was detected in whole living cells using flow cytometry and in paraffin-embedded tumor tissues using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results The triple-regulated CRAd carrying SG611-PDCD5 and nude mouse xenograft models of K562 cells were successfully constructed. In vitro treatment with SG611-PDCD5 in combination with low-dose daunorubicin elicited more potent anti-proliferative and proapoptotic effects in leukemic cells in a dose-dependent manner. The Chou-Talalay analysis revealed synergistic anti-proliferative effects in all of the above cell lines. In the nude mice xenograft model, the tumor size in the control, daunorubicin, SG611-PDCD5, and combined treatment groups on day 10 were 170.1±47.8, 111.9±81.1, 60.7±12.3, and 33.2±17.5 mm3, respectively (all P<0.05). The results of the TUNEL assay showed significantly more apoptotic cells in the SG611-PDCD5 plus daunorubicin group than in the SG611-PDCD5 or daunorubicin groups alone (25±0.82, 12.5±2.27, and 7.8±2.67 apoptotic cells/field, respectively) (P<0.05). Conclusion The findings suggest that combined treatment with SG611-PDCD5 and daunorubicin may be a promising strategy for enhancing chemosensitivity and thus lowering the dose-related toxicity of daunorubicin in leukemia therapy.