National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.
BMC Cancer. 2011 Feb 3;11:54. doi: 10.1186/1471-2407-11-54.
Adeno-associated virus-2 (AAV-2)-mediated gene therapy is quite suitable for local or regional application in head and neck cancer squamous cell carcinoma (HNSCC). However, its low transduction efficiency has limited its further development as a therapeutic agent. DNA damaging agents have been shown to enhance AAV-mediated transgene expression. Cisplatin, one of the most effective chemotherapeutic agents, has been recognized to cause cancer cell death by apoptosis with a severe toxicity. This study aims to evaluate the role of cisplatin in AAV-mediated tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) expression and the effect on HNSCC both in vitro and in vivo.
Five human HNSCC cell lines were treated with recombinant soluble TRAIL (rsTRAIL) and infected with AAV/TRAIL to estimate the sensitivity of the cancer cells to TRAIL-induced cytotoxicity. KB cells were infected with AAV/EGFP with or without cisplatin pretreatment to evaluate the effect of cisplatin on AAV-mediated gene expression. TRAIL expression was detected by ELISA and Western blot. Cytotoxicity was measured by MTT assay and Western blot analysis for caspase-3 and -8 activations. Following the in vitro experiments, TRAIL expression and its tumoricidal activity were analyzed in nude mice with subcutaneous xenografts of HNSCC.
HNSCC cell lines showed different sensitivities to rsTRAIL, and KB cells possessed both highest transduction efficacy of AAV and sensitivity to TRAIL among five cell lines. Preincubation of KB cells with subtherapeutic dosage of cisplatin significantly augmented AAV-mediated transgene expression in a heparin sulfate proteoglycan (HSPG)-dependent manner. Furthermore, cisplatin enhanced the killing efficacy of AAV/TRAIL by 3-fold on KB cell line. The AAV mediated TRAIL expression was observed in the xenografted tumors and significantly enhanced by cisplatin. AAV/TRAIL suppressed the tumors growth and cisplatin augmented the tumoricidal activity by two-fold. Furthermore, Combination treatment reduced cisplatin-caused body weight loss in nude mice.
The combination of AAV-mediated TRAIL gene expression and cisplatin had synergistic therapeutic effects on head and neck cancers and reduced the potential toxicity of cisplatin. These findings suggest that the combination of AAV/TRAIL and cisplatin may be a promising strategy for HNSCC therapy.
腺相关病毒-2（AAV-2）介导的基因治疗非常适合头颈部鳞状细胞癌（HNSCC）的局部或区域性应用。然而，其转导效率低限制了其作为治疗剂的进一步发展。已证实 DNA 损伤剂可增强 AAV 介导的转基因表达。顺铂是最有效的化疗药物之一，已被证实通过细胞凋亡导致癌细胞死亡，但其毒性严重。本研究旨在评估顺铂在 AAV 介导的肿瘤坏死因子相关凋亡诱导配体（TRAIL）表达中的作用及其对体外和体内 HNSCC 的影响。
用重组可溶性 TRAIL（rsTRAIL）处理五个人类 HNSCC 细胞系并感染 AAV/TRAIL，以评估癌细胞对 TRAIL 诱导的细胞毒性的敏感性。KB 细胞用 AAV/EGFP 感染，或先用顺铂预处理，以评估顺铂对 AAV 介导的基因表达的影响。通过 ELISA 和 Western blot 检测 TRAIL 表达。通过 MTT 测定和 caspase-3 和 -8 激活的 Western blot 分析评估细胞毒性。在体外实验之后，分析裸鼠皮下移植 HNSCC 中的 TRAIL 表达及其杀瘤活性。
HNSCC 细胞系对 rsTRAIL 的敏感性不同，KB 细胞在五细胞系中具有最高的 AAV 转导效率和对 TRAIL 的敏感性。KB 细胞用亚治疗剂量的顺铂预孵育可显著增强 HSPG 依赖性 AAV 介导的转基因表达。此外，顺铂使 AAV/TRAIL 对 KB 细胞系的杀伤效力提高了 3 倍。在异种移植瘤中观察到 AAV 介导的 TRAIL 表达，并被顺铂显著增强。AAV/TRAIL 抑制肿瘤生长，顺铂使杀瘤活性增强两倍。此外，联合治疗减少了顺铂引起的裸鼠体重减轻。
AAV 介导的 TRAIL 基因表达与顺铂联合治疗对头颈部癌症具有协同治疗作用，并降低了顺铂的潜在毒性。这些发现表明，AAV/TRAIL 与顺铂的联合可能是治疗 HNSCC 的一种有前途的策略。
