Department of Medical Biology and Genetics, Human Gene and Cell Therapy Center of Akdeniz University Hospitals and Clinics, Antalya 07058, Turkiye.
BMC Cancer. 2010 Oct 27;10:584. doi: 10.1186/1471-2407-10-584.
Lung cancer causes the highest rate of cancer-related deaths both in men and women. As many current treatment modalities are inadequate in increasing patient survival, new therapeutic strategies are required. TNF-related apoptosis-inducing ligand (TRAIL) selectively induces apoptosis in tumor cells but not in normal cells, prompting its current evaluation in a number of clinical trials. The successful therapeutic employment of TRAIL is restricted by the fact that many tumor cells are resistant to TRAIL. The goal of the present study was to test a novel combinatorial gene therapy modality involving adenoviral delivery of TRAIL (Ad5hTRAIL) and IKK inhibition (AdIKKβKA) to overcome TRAIL resistance in lung cancer cells.
Fluorescent microscopy and flow cytometry were used to detect optimum doses of adenovirus vectors to transduce lung cancer cells. Cell viability was assessed via a live/dead cell viability assay. Luciferase assays were employed to monitor cellular NF-κB activity. Apoptosis was confirmed using Annexin V binding.
Neither Ad5hTRAIL nor AdIKKβKA infection alone induced apoptosis in A549 lung cancer cells, but the combined use of Ad5hTRAIL and AdIKKβKA significantly increased the amount of A549 apoptosis. Luciferase assays demonstrated that both endogenous and TRAIL-induced NF-κB activity was down-regulated by AdIKKβKA expression.
Combination treatment with Ad5hTRAIL and AdIKKβKA induced significant apoptosis of TRAIL-resistant A549 cells, suggesting that dual gene therapy strategy involving exogenous TRAIL gene expression with concurrent IKK inhibition may be a promising novel gene therapy modality to treat lung cancer.
肺癌是导致男性和女性癌症相关死亡的主要原因。由于许多当前的治疗方法并不能提高患者的生存率，因此需要新的治疗策略。肿瘤坏死因子相关凋亡诱导配体（TRAIL）选择性地诱导肿瘤细胞凋亡，而不诱导正常细胞凋亡，这促使其在许多临床试验中得到评估。TRAIL 的成功治疗应用受到许多肿瘤细胞对 TRAIL 耐药的限制。本研究的目的是测试一种新的联合基因治疗模式，涉及腺病毒递送 TRAIL（Ad5hTRAIL）和 IKK 抑制（AdIKKβKA），以克服肺癌细胞中的 TRAIL 耐药性。
荧光显微镜和流式细胞术用于检测转导肺癌细胞的最佳剂量的腺病毒载体。通过死活细胞活力测定法评估细胞活力。使用荧光素酶测定法监测细胞 NF-κB 活性。通过 Annexin V 结合来确认细胞凋亡。
单独感染 Ad5hTRAIL 或 AdIKKβKA 均不能诱导 A549 肺癌细胞凋亡，但 Ad5hTRAIL 和 AdIKKβKA 的联合使用显著增加了 A549 细胞凋亡的数量。荧光素酶测定法表明，内源性和 TRAIL 诱导的 NF-κB 活性均被 AdIKKβKA 表达下调。
Ad5hTRAIL 和 AdIKKβKA 的联合治疗显著诱导 TRAIL 耐药的 A549 细胞凋亡，表明外源性 TRAIL 基因表达与同时进行的 IKK 抑制的双重基因治疗策略可能是治疗肺癌的一种有前途的新基因治疗模式。
