Wang Shu, Liu Xiaoqun, Qiao Tiankui, Zhang Qi
Department of Oncology, Jinshan Hospital, Medical Centre of Fudan University, Jinshan District, Shanghai, China.
J Int Med Res. 2017 Dec;45(6):2009-2022. doi: 10.1177/0300060517728634. Epub 2017 Sep 18.
Objective To evaluate the radiosensitivity effect of CpG oligodeoxyribonucleotide (ODN) 7909 on human epidermoid cancer strain-2 (Hep-2) cells in vitro and discuss the potential for improved radiotherapy treatment in patients with laryngeal squamous cell carcinoma. Methods Toll-like receptor ( TLR) 9 expression was assessed in Hep-2 cells using Western blots and reverse transcription polymerase chain reaction. Cell Counting Kit-8 was used to detect Hep-2 cell viability at 24 and 48 h following treatment with different CpG ODN7909 concentrations. Cellular colonization was evaluated using microscopy. Cell cycle distribution and apoptosis rate was determined with flow cytometry. Interleukin (IL)-12 and tumour necrosis factor (TNF)-α concentrations were detected by enzyme-linked immunosorbent assay. Results Hep-2 cells were found to express TLR9, and CpG ODN7909 treatment suppressed Hep-2 cell viability in a dose- and time-dependent manner. Cell survival curve analyses revealed a sensitivity enhancement ratio of the mean death dose of 1.225 for CpG ODN7909 plus irradiation versus irradiation alone. Furthermore, the population of Gap 2/mitotic-phase cells, apoptosis rate and secreted IL-12 and TNF-α levels were significantly increased in Hep-2 cells treated with CpG ODN7909 plus irradiation versus IR alone. Conclusion CpG ODN7909 enhanced the radiosensitivity of Hep-2 cells in vitro.
目的 评估CpG寡脱氧核苷酸（ODN）7909对人表皮样癌2株（Hep-2）细胞的体外放射增敏作用，并探讨其改善喉鳞状细胞癌患者放射治疗效果的潜力。方法 采用蛋白质免疫印迹法和逆转录聚合酶链反应评估Hep-2细胞中Toll样受体（TLR）9的表达。使用细胞计数试剂盒-8检测不同浓度CpG ODN7909处理24和48小时后Hep-2细胞的活力。通过显微镜评估细胞集落形成情况。用流式细胞术测定细胞周期分布和凋亡率。采用酶联免疫吸附测定法检测白细胞介素（IL）-12和肿瘤坏死因子（TNF）-α的浓度。结果 发现Hep-2细胞表达TLR9，且CpG ODN7909处理以剂量和时间依赖性方式抑制Hep-2细胞活力。细胞存活曲线分析显示，与单纯照射相比，CpG ODN7909联合照射的平均死亡剂量的增敏比为1.225。此外，与单纯照射相比，CpG ODN7909联合照射处理的Hep-2细胞中G2/有丝分裂期细胞比例、凋亡率以及分泌的IL-12和TNF-α水平均显著增加。结论 CpG ODN7909在体外增强了Hep-2细胞的放射敏感性。
