CAJ | 학술논문

目的：通过人IFN-λ1重组腺病毒（ r-Ad-hIFN-λ1）对人类胃癌裸鼠原位移植瘤模型的干预，探讨人r-Ad-hIFN-λ1重组腺病毒对胃癌生长的抑制作用及其部分机制。方法应用原位移植方法建立裸鼠人胃癌原位移植瘤模型，后随机分为3组：PBS对照组、Ad-Lac Z空载体组、r-Ad-hIFN-λ1实验组。每周干预1次，共3次，不同时间点二维B超探查裸鼠原位肿瘤的瘤体大小并绘制生长曲线。末次干预后1周处死，收集相应组织， RT-PCR检测骨骼肌中IFN-λ1 mRNA的表达；Western blot检测胃癌组织中IFN-λ1蛋白的表达；免疫组化染色检测肿瘤组织切片中IFN-λ1的表达；TUNEL法检测瘤体石蜡切片组织凋亡；流式细胞仪分析脾中NK细胞比例。结果实验组裸鼠胃癌移植瘤体积（248．39±7．60） mm3明显小于对照组（322．92±5．92） mm3和空载体组（331．25±6．00） mm3（P＜0．05）；hIFN-λ1成功转染骨骼肌并在胃癌组织中表达；免疫组化切片中实验组肿瘤组织胞质内hIFN-λ1强阳性表达；TUNEL测凋亡指数示实验组有大片凋亡（0．700±0．059）明显区别于对照组（0．271±0．026）和空载体组（0．333±0．028）（P＜0．05）；实验组裸鼠脾脏中NK细胞比例（26．49±1．89）％明显大于对照组（13．94±1．31）％及空载体组（19．12±1．69）％（P＜0．05）。结论 r-Ad-hIFN-λ1通过骨骼肌转染能够显著抑制胃癌生长，并可能通过诱导其凋亡、刺激NK细胞而起作用。
목적：통과인IFN-λ1중조선병독（ r-Ad-hIFN-λ1）대인류위암라서원위이식류모형적간예，탐토인r-Ad-hIFN-λ1중조선병독대위암생장적억제작용급기부분궤제。방법응용원위이식방법건립라서인위암원위이식류모형，후수궤분위3조：PBS대조조、Ad-Lac Z공재체조、r-Ad-hIFN-λ1실험조。매주간예1차，공3차，불동시간점이유B초탐사라서원위종류적류체대소병회제생장곡선。말차간예후1주처사，수집상응조직， RT-PCR검측골격기중IFN-λ1 mRNA적표체；Western blot검측위암조직중IFN-λ1단백적표체；면역조화염색검측종류조직절편중IFN-λ1적표체；TUNEL법검측류체석사절편조직조망；류식세포의분석비중NK세포비례。결과실험조라서위암이식류체적（248．39±7．60） mm3명현소우대조조（322．92±5．92） mm3화공재체조（331．25±6．00） mm3（P＜0．05）；hIFN-λ1성공전염골격기병재위암조직중표체；면역조화절편중실험조종류조직포질내hIFN-λ1강양성표체；TUNEL측조망지수시실험조유대편조망（0．700±0．059）명현구별우대조조（0．271±0．026）화공재체조（0．333±0．028）（P＜0．05）；실험조라서비장중NK세포비례（26．49±1．89）％명현대우대조조（13．94±1．31）％급공재체조（19．12±1．69）％（P＜0．05）。결론 r-Ad-hIFN-λ1통과골격기전염능구현저억제위암생장，병가능통과유도기조망、자격NK세포이기작용。
Objective To investigate the inhibitory effects of a recombinant adenovirus vector ex-pressing human IFN-λ1 ( r-Ad-hIFN-λ1) on the growth of orthotopic gastric cancer and to analyze its possi-ble mechanism in a nude mice model of orthotopic human gastric carcinoma .Methods Orthotopic trans-plantation was performed to establish the nude mice model of orthotopic gastric cancer .Thirty mice were ran-domly divided into three groups including PBS control group , Ad-Lac Z empty vector group and r-Ad-hIFN-λ1 experiment group .The mice in each group were given the corresponding interventions once a week for three times.The sizes of tumor were detected by using B-mode ultrasound at different time points to draw growth curves .The expression of IFN-λ1 at mRNA level in skeletal muscle tissues was analyzed by RT-PCR.Western blot assay and immunohistochemical staining were used to detect IFN-λ1 protein in gastric tumor samples .The apoptosis of cells in paraffin-embedded tumor tissues was detected by TUNEL .The per-centages of natural killer ( NK) cells in spleen tissues were analyzed by flow cytometry .Results Compared with control group and empty vector group , the mice in r-Ad-hIFN-λ1 experiment group showed the smallest tumor size [(331.25±6.00) mm3, (322.92±5.92) mm3 vs (248.39±7.60) mm3; P<0.05].hIFN-λ1 was transfected into skeletal muscle successfully and expressed in gastric cancer tissue .Highly expressed hIFN-λ1 was observed in cytoplasm of tumor cells from experiment group by immunohistochemical staining . The apoptotic index of tumor tissues for experiment group was 0.700±0.059 which was significant different from that of control group (0.271±0.026, P<0.05) and empty vector group (0.333±0.028, P<0.05). The percentage of NK cells in spleens from mice in experiment group [(26.49±1.89)%] was significantly higher than that of control group [(13.94 ±1.31)%, P<0.05)] and empty vector group [(19.12 ±1.69)%, P<0.05)].Conclusion Transfection of r-Ad-hIFN-λ1 and expression of hIFN-λin skeletal muscle could significantly inhibit the growth of gastric cancer by inducing tumor cells ′apoptosis and enhan-cing NK cells.