Bu Xu-Feng, Zhang Jie, Jia Li-Juan, Liang Bing, Zhang Jin, Liu Yang, Yan Yu-Lan
Department of General Surgery, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu 212002, P.R. China.
Department of General Surgery, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu 212002, P.R. China ; Clinical Medicine College of Jiangsu University, Zhenjiang, Jiangsu 212002, P.R. China.
Exp Ther Med. 2014 Oct;8(4):1115-1122. doi: 10.3892/etm.2014.1896. Epub 2014 Aug 11.
The aim of the present study was to investigate the effect of human interferon-λ1 recombinant adenovirus (r-Ad-hIFN-λ1) on gastric carcinoma. Human SGC-7901 cells were utilized to create an orthotopic implantation model of gastric cancer in nude mice through sterile surgery. The mice were randomly divided into three groups: Phosphate-buffered saline control (blank), adenovirus encoding bacterial β-galactosidase (Ad-Lac Z) empty vector and r-Ad-hIFN-λ1. Tumor size was measured every seven days. After three weeks of treatment, the tumors in the mice were detected by abdominal B ultrasound. The cDNA of IFN-λ1 expression in skeletal muscle was detected by a reverse transcription polymerase chain reaction and IFN-λ1 protein expression in the tumors was detected by western blot analysis and immunohistochemistry. Flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays were conducted to analyze the proportion of natural killer (NK) cells in the spleen and the rate of cell apoptosis in tumor paraffin sections. Prior to sacrifice, the size of the tumors in the r-Ad-hIFN-λ1, Ad-Lac Z and blank groups was 184.29±10.84 mm, 234.62±10.59 mm and 253.18±7.69 mm, respectively (P<0.001). The lymph node metastasis in the abdominal cavity was 0% in the r-Ad-hIFN-λ1 group, 50% in the Ad-Lac Z group and 80% in the blank group (P<0.005). Furthermore, IFN-λ1 mRNA and protein were highly expressed in the r-Ad-hIFN-λ1 group, and the apoptosis rate in the r-Ad-hIFN-λ1 group was higher than that in the Ad-Lac Z and blank groups. The proportion of NK cells in the spleens of nude mice in the r-Ad-hIFN-λ1, Ad-Lac Z and blank groups was 26.53±1.54, 17.70±1.09 and 16.35±1.43%, respectively (P<0.001). The TUNEL results showed there was significantly more severe apoptosis in the r-Ad-hIFN-λ1 group than that in the two other groups. The apoptosis indices in the r-Ad-hIFN-λ1, Ad-Lac Z and blank groups were 0.772±0.075, 0.329±0.169 and 0.265±0.049, respectively. In conclusion, the r-Ad-hIFN-λ1 significantly inhibited human gastric cancer, possibly by promoting apoptosis of the tumors and stimulating immunological function.
本研究旨在探讨人干扰素λ1重组腺病毒（r-Ad-hIFN-λ1）对胃癌的影响。通过无菌手术，利用人SGC-7901细胞在裸鼠体内建立胃癌原位植入模型。将小鼠随机分为三组：磷酸盐缓冲盐水对照组（空白组）、编码细菌β-半乳糖苷酶的腺病毒（Ad-Lac Z）空载体组和r-Ad-hIFN-λ1组。每七天测量一次肿瘤大小。治疗三周后，通过腹部B超检测小鼠体内的肿瘤。通过逆转录聚合酶链反应检测骨骼肌中IFN-λ1表达的cDNA，通过蛋白质免疫印迹分析和免疫组织化学检测肿瘤中IFN-λ1蛋白的表达。进行流式细胞术和末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记（TUNEL）分析，以分析脾脏中自然杀伤（NK）细胞的比例以及肿瘤石蜡切片中的细胞凋亡率。处死前，r-Ad-hIFN-λ1组、Ad-Lac Z组和空白组的肿瘤大小分别为184.29±10.84 mm、234.62±10.59 mm和253.18±7.69 mm（P<0.001）。r-Ad-hIFN-λ1组腹腔淋巴结转移率为0%，Ad-Lac Z组为50%，空白组为80%（P<0.005）。此外，r-Ad-hIFN-λ1组中IFN-λ1 mRNA和蛋白高表达，且r-Ad-hIFN-λ1组的凋亡率高于Ad-Lac Z组和空白组。r-Ad-hIFN-λ1组、Ad-Lac Z组和空白组裸鼠脾脏中NK细胞的比例分别为26.53±1.54%、17.70±1.09%和16.35±1.43%（P<0.001）。TUNEL结果显示，r-Ad-hIFN-λ1组的凋亡明显比其他两组严重。r-Ad-hIFN-λ1组、Ad-Lac Z组和空白组的凋亡指数分别为0.772±0.075、0.329±0.169和0.265±0.049。总之，r-Ad-hIFN-λ1可显著抑制人胃癌，可能是通过促进肿瘤细胞凋亡和刺激免疫功能实现的。
