转染IL-17基因的小鼠结肠癌细胞体内抗肿瘤机制研究  

Antitumor mechanism of mouse colon cancer cells transfected with IL-17 gene in vivo

作　　者：李艳双[1] 宋晓天[2] 张征峥[2] 钱雪松[2] 刘伟[2] 杨丽娟[2] 

机构地区：[1]河北联合大学临床医学院转化医学实验室,唐山063000 [2]河北医科大学基础医学院免疫教研室,石家庄050017

出　　处：《中国免疫学杂志》2015年第5期643-649,662,共8页Chinese Journal of Immunology

基　　金：河北省教育厅基金(No.2011101)资助

摘　　要：目的:研究IL-17体内抗肿瘤的作用及其机制,为IL-17基因疫苗进入临床提供实验依据。方法:利用已建立的稳定转染小鼠IL-17全长基因的小鼠结肠癌细胞(C26/pc DNA3.1-IL-17)及相应的对照细胞(C26/pc DNA3.1、C26)建立荷瘤动物模型,观察小鼠的成瘤性、肿瘤生长情况及小鼠生存期的变化,检测各组小鼠脾细胞增殖情况、脾NK细胞杀伤活性;及脾淋巴细胞中Th1、Th2、Th17、Treg细胞特征性细胞因子和/或转录因子的表达;检测各组小鼠肿瘤浸润的淋巴细胞(TIL)的增殖反应情况;及肿瘤浸润巨噬细胞中M1特征性细胞因子IL-10和M2特征性细胞因子IL-12的表达。结果:将C26/pc DNA3.1-IL-17细胞接种小鼠体内后,荷瘤小鼠的肿瘤生长速度、体积均明显小于C26/pc DNA3.1细胞组及C26细胞组(P<0.05),接种C26/pc DNA3.1-IL-17细胞的雄性小鼠移植瘤明显小于雌性小鼠移植瘤体积(P<0.05),各组小鼠生存时间没有明显区别(P>0.05);接种C26/pc DNA3.1-IL-17细胞的小鼠脾细胞较接种C26、C26/pc DNA3.1细胞的小鼠脾细胞增殖能力强(P<0.05),与正常小鼠脾细胞增殖能力比较无明显统计学差异(P>0.05),接种C26、C26/pc DNA3.1细胞的小鼠脾细胞增殖能力较正常小鼠脾细胞增殖能力降低(P<0.05);在效靶比为40∶1和20∶1时,接种三种细胞的小鼠NK杀伤率较正常小鼠均降低(P<0.05),在效靶比40∶1时,接种C26/pc DNA3.1-IL-17细胞比接种C26、C26/pc DNA3.1细胞的小鼠的脾淋巴细胞的NK杀伤率明显增强(P<0.05),效靶比10∶1时,各组之间没有明显统计学差异(P>0.05);接种C26/pc DNA3.1-IL-17细胞的小鼠脾淋巴细胞较接种C26、C26/pc DNA3.1的小鼠脾淋巴细胞表达更高水平的IFN-γ(Th1的特征性细胞因子)、IL-4(Th2特征性细胞因子)、GATA-3(Th2特征性转录因子)、ROR-γt(Th17特征性转录因子)、IL-10(Treg特征性细胞因子)mRNA(P<0.05);接种C26/pc DNA3.1-IL-17细胞的小鼠TIL较接种C26、C26/pc DNA3.1细胞的小鼠TIL�Objective：To investigate the effects of interleukin-17 on tumor,we transfected interleukin-17 gene into mouse colon cancer cells（C26） and set up an animal model in tumor. Methods： By plasmid vector, IL-17 gene was transfected into C26. Meanwhile, empty plasmid vector（ pcDNA3.1 ） and C26 cells were transfected as control groups. C26/pcDNA3. 1-IL-17, C26/pcDNA3. 1, and C26 cells were subcutaneously inoculated into mice respectively and the tumor volume and the survival time were observed. Proliferation of splenocyte and NK activity were detected. Detect the characteristic cytokines and transcriptional factors of Thl ,Th2 ,Thl7 and Treg cells in splenic lymphocyte. Proliferation of TIL was detected. The characteristic cytokines IL-IO of M1 and the characteristic cytokines IL-12 of M2 in tumor infiltrating macrophages were detected. Results： The growth of tumor in mice inoculated with C26/pcDNA3. 1-IL-17 cells was significantly retarded（ P〈O. 05）, and the growth of tumor in male mice inoculated with C26/pcDNA3. 1-IL-17 cells was significantly retarded than female mice （ P 〈 0. 05 ）. The mice survival time of C26/pcDNA3. 1-IL-17 group was similar with C26/ pcDNA3.1 and C26 groups（P〉0. 05）. The proliferation of the splenocytes from C26/pcDNA3.1-IL-17 inoculated mice was higher than those of C26/pcDNA3.1, C26 groups（P〈0. 05 ）, but was similar with the normal group（ P〉O. 05 ）, the proliferation of the splenocytes from C26/pcDNA3.1 and C26 inoculated mice was slow than those of normal groups（ P〈0. 05 ）. The NK（ separate from spleen） activity of C26/pcDNA3.1-IL-17, C26/pcDNA3.1 and C26 inoculated mice was lower than those of normal groups when the ratios of effector ceils and target cells were 40：1,20：1 （P〈0. 05 ） ,the NK（ separate from spleen） activity of C26/pcDNA3.1-IL-17 inoculated mice was higher than those of C26/pcDNA3.1 and C26 groups when the ratios of effector cells and target cells were 40：1 （P〈0. 05）, there＇s no difference among every

关 键 词：IL-17 小鼠结肠癌细胞株C26 抗肿瘤作用 

分 类 号：R392.12[医药卫生—免疫学]