髓系细胞RelA／p65基因介导吸烟促进小鼠肺癌的生长机制  

Mechanisms of myeloid cell RelA/p65 in cigarette smoking-induced lung cancer growth in mice

作　　者：姚懿雯[1] 吴军录[1] 权文强[1] 周宏[1] 张宇[1] 万海英[1] 李冬[1] 

机构地区：[1]同济大学附属同济医院检验科,上海200065

出　　处：《中华肿瘤杂志》2014年第6期412-417,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金面上项目（81272603）;上海市浦江人才计划（13PJ4407300）

摘　　要：目的研究吸烟促进小鼠肺癌生长的作用机制。方法使用髓系细胞RelA／p65敲除小鼠（RelA／p65^-/-）和对照WT小鼠建立小鼠转移性肺癌模型，分为吸烟组和空气组，计数小鼠肺表面肿瘤数并对肿瘤大小进行评价，利用Kaplan—Meier生存曲线分析小鼠的生存情况。采用免疫组化法检测小鼠肿瘤增殖抗原Ki-67、肿瘤坏死因子仅（TNF-α）和CD68的表达，采用Westernblot法检测肿瘤细胞cyclinDI和c-myc的表达，采用原位末端转移酶标记技术（TUNEL）检测肿瘤细胞的凋亡，采用酶联免疫吸附（ELISA）试验检测肺肿瘤组织中TNF-α、白细胞介素6（IL-6）和KC的浓度。结果WT小鼠在受到香烟刺激后，肺重量、肿瘤个数和最大肿瘤直径分别为（1．5±0．1）g、（64．8±4．1）个和（7．6±0．2）mm，均明显高于未受香烟刺激的WT小鼠（均P〈0．05）；而RelA／p65^-/-小鼠的肺重量、肿瘤个数和最大肿瘤直径在香烟刺激前后无明显变化（均P〉0．05）。生存分析的结果显示，WT吸烟组小鼠的生存时间明显低于WT空气组（P〈0．05），但RelA／p65^-/-小鼠的生存时间均明显长于WT小鼠（均P〈0．05）。WT空气组、WT吸烟组、RelA／p65^-/-空气组和RelA／p65^-/-吸烟组小鼠Ki-67阳性肿瘤细胞的百分比分别为（43．4±2．9）％、（60．6±5．4）％、（12．8±3．6）％和（15．0±4．2）％，香烟刺激后，WT小鼠Ki-67阳性肿瘤细胞百分比明显增高（P〈0．05），而RelA／p65^-/-小鼠Ki-67的表达无明显变化（P〉0．05）。WT空气组、WT吸烟组、RelA／p65^-/-空气组和RelA／p65^-/-吸烟组小鼠肿瘤细胞的凋亡率分别为（11．6±1．7）％、（13．0±2．0）％、（13．2±2．0）％和（11．0±1．4）％，差异均无统计学意义（均P〉0．05）。WT小鼠受到香烟刺激后，肿瘤细胞中cyclinD1和c—myc蛋白的表达明显增强；而RelA／p65^-/-小鼠受到香烟刺激后�Objective The aim of this study was to investigate the mechanism of cigarette smoking （CS）-induced lung cancer growth in mice. Methods RelA/p65^-/- mice and WT mice were used to establish mouse models of lung cancer. Both mice were divided into two groups： air group and CS group, respectively. Tumor number on the lung surface was counted and maximal tumor size was evaluated using HE staining. Kaplan Meier （K-M） survival curve was used to analyze the survival rate of the mice. Expression of Ki-67, TNF-α and CD68 in the tumor tissue was determined by immunohistoehemical analysis, and cyclin D1 and c-myc proteins were examined by Western blot. Apoptosis of tumor cells was analyzed using TUNEL staining. The concentrations of inflammatory eytokines TNF-α, IL-6 and KC in the mouse lung tissues were evaluated by ELISA. Results Compared with the WT air group, the lung weight, lung tumor muhiplieity, as well as maximum tumor size in the WT mice exposed to CS were （1.5 ±0. 1）g, （64.8 ±4. 1） and （7.6 ±0.2） mm, respectively, significantly increased than those in the WT mice not exposed to CS （ P 〈 0.05 for all）. However, there were no statistically significant differences between RelA/p65^-/- mice before and after CS exposure （ P 〉 0. 05 for all）. Kaplan-Meier survival analysis showed that CS exposure significantly shortened the life time of WT mice （ P 〈 0.05 ）, and deletion of RelA/p65 in myeloid cells resulted in an increased survival compared with that of the WT mice （P 〈 0.05 for all）. The ratios of Ki-67 positive tumor cells were （43.4± 2.9） %, （ 60.6 ± 5.4） % , （ 12.8 ±3.6） % and （ 15.0 ± 4.2） % in the WT air group, WT CS groups, RelA/p65^-/- air groups and Re1A/p65^-/- CS groups, respectively. After smoking, the number of Ki-67-positive cells was significantly increased in the WT mice （ P 〈 0.05 ）. However, there was no significant difference between the RelA/p65^-/- groups before and after smoking （P 〉 0. 05 ）. The apoptosis rate of W

关 键 词：吸烟 肺肿瘤 肿瘤生长 核因子κB RELA P65基因 细胞凋亡 小鼠 

分 类 号：R734.2[医药卫生—肿瘤]