摘要
目的:近年来,代谢与肿瘤发病的病因学研究已经成为热点,同时肺癌仍居恶性肿瘤发病率和死亡率榜首,本研究拟构建一个稳定易操作的肺癌原位移植瘤模型,并初步探讨代谢相关通路在该原位移植瘤模型中的表达。方法:选取雄性6~8周龄BABL/c裸鼠10只,优化麻醉用药及手术操作步骤,经气管移植5×106个细胞的A549肺癌细胞成瘤,观察移植瘤形成率、形成部位、大体形态、增殖分化程度。另取4只相同条件裸鼠进行手术空白对照。同时检测代谢相关通路蛋白STAT3、AKT和m TOR的表达情况等。结果:手术最佳麻醉条件为75 mg/kg戊巴比妥钠腹腔注射;改进操作后经气管移植的小鼠肺癌形成率为80.0%;肿瘤形成以右肺上叶居多,呈明显腺癌结构,分化程度较低,移植瘤组织增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达高于肺组织,呈高表达状态。肿瘤负荷肺组织代谢相关通路蛋白信号传导与转录激活因子3(signal transducer and activator of transcription 3,STAT3)、蛋白激酶B(protein kinase B,PKB即AKT)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,m TOR)的磷酸化水平即p STAT3/STAT3、p AKT/AKT、p-m TOR/m TOR比值均高于正常肺组织,差异具有统计学意义(t=-4.381,-4.462,-4.951;P=0.041,0.011,0.038)。结论:通过改良经气管移植手术方式,可以构建一种操作简单、低成本、高效稳定的A549细胞肺癌移植瘤模型;代谢相关通路在肺癌的发生发展中也发挥了一定作用。
Objective ：To build a moderate lung cancer orthotopic transplantation tumor model, and to investigate the expression of metabolic pathway in the this model. Methods：After optimizing the dose of anesthetic medicine and operation procedures of endotracheal orthotopic transplantation,5×10^6 of A549 cells were planted into male nude mice（6-8 weeks old,n=10） lungs. And the blank control （only PBS transplant） was carry out on the same condition mice（n=4）. Then the tumor formation rate,location,anatomy and the ex- pression of signal transducer and activator of transcription 3 （STAT3）,protein kinase B （AKT） and mammalian target of rapamycin （mTOR） were studied. Results：The best surgical anaesthesia condition was by intraperitoneal injection of 75 mg/kg body weight pentobarbital sodium. After the improvement of the operation procedure, the total tumor formation rate was 80.0%, and mainly located in the upper lobe of the right lung. The tumor showed a significant glandular cancer structure with poor differentiation degree,and more proliferating cell nuclear antigen（PCAN）-positive cells compared with normal lungs. The levels of phosphorylation of STAT3, AKT and mTOR and the ratio of pSTAT3/STAT3,p-AKT/AKT,p-mTOR/mTOR in lung tumors were significantly higher than those in normal lung tissues, the difference was statistically significant differences （ t = -4.381, -4.462, -4.951 ; P=0.041,0.011,0.038 respectively ）.Conclusion：The improved endotracheal orthotopic transplanta- tion lung cancer model is a simple,low cost,stable model,and metabolic pathways play a role in the development and progre- ssion of lung cancer.
出处
《重庆医科大学学报》
CSCD
北大核心
2017年第7期888-892,共5页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:81071907)
教育部新世纪优秀人才支持计划资助项目(编号:NCET-10-0997)
