EL4细胞STAT4沉默前后培养上清对小鼠肿瘤生长的影响  

Impact of Supernatant of EL4 Cells with STAT4(-)/STAT4 (+) on Mice Bearing H22 Liver Cancer

作　　者：魏敏[1] 南英[1] 杨松[1] 陈名珍[1] 魏晓丽[1] 王青山[1] 李玉皓[1] 向荣[1] 王悦[1] 

机构地区：[1]南开大学医学院,300071

出　　处：《天津医药》2009年第11期948-951,I0003,共5页Tianjin Medical Journal

基　　金：南开大学本科生科研基金资助项目;863国家高技术研究发展计划项目(项目编号:2007AA021010);973国家重大科学研究计划资助项目(项目编号:2007CB914804);天津市应用基础及前沿技术研究计划(项目编号:08JCYBJC07100)

摘　　要：目的:探讨沉默小鼠T淋巴瘤细胞(EL4细胞)信号转导和转录激活因子4(STAT4)前后,其改变的培养上清液成分对H22肝癌荷瘤小鼠腹水生长及个体生存状况的影响。方法:应用ELISA方法检测肿瘤细胞STAT4沉默后培养上清液中白介素(IL)-10水平;将H22肝癌荷瘤小鼠随机分为STAT4(+)组、STAT4(-)组、IL-10组、培养基组,分别腹腔注射STAT4沉默前、后培养细胞上清液,IL-10,1640培养基,检测荷瘤小鼠体质量、腹围、腹水、生存率及腹水瘤细胞存活等指标;RT-PCR半定量检测荷瘤小鼠脾细胞的Foxp3mRNA表达。结果:STAT4沉默后培养细胞上清液IL-10水平明显增高;用不同的培养上清液成分注射后STAT4(-)组体质量增长率较STAT4(+)组显著降低,与培养基组各指标比较差异无统计学意义;15d内生存率比较,STAT4(-)组显著高于STAT4(+)组、IL-10组;STAT4(-)组脾细胞Foxp3表达水平高于其他3组。结论:STAT4沉默后培养细胞上清液局部注射荷瘤小鼠,能抑制癌性腹水增长,显著提高存活率,但脾细胞Foxp3mRNA水平明显增高,可抑制荷瘤小鼠的抗肿瘤免疫,这与近期观察到的结果相悖。Objective： To investigate the impact of supernatant of EL4 cells with STAT4 negative expression （STAT4-） and with STAT4 normal expression （STAT4＋） on cancerous ascites and survival condition of mice bearing H22 liver cancer. Methods： The levels of IL-10 were detected in EL4 cells with STAT4 （＋）/STAT4 （-） by ELISA. The mice beating H22 liver cancer were randomized into 4 groups： STAT4（＋） group, STAT4（-） group, IL-10 group, and medium control group. The super- natant of EIA cells with STAT4-, supernatant of EIA cells with STAT4 （＋）, IL-10 and 1640 medium were administrated in- traperitoneally, respectively. The body mass （BM）, abdomen circumference （AC）, volume of ascites, survival rate and survival rate of fresh ascetic tumor cells were detected in groups. The level of Foxp3 mRNA in extracted spleen cells of mice beating cancer was detected via RT-PCR. Results： The level of IL-10 was higher in EL4 cells with STAT4（-） than that of STAT4（＋） in EL4 cells. The BM growth rate was significantly decreased in STAT4（-） group than that of STAT4（＋） group（P〈0.05） after the supernatant treatment. The 15-day survival rate was higher in STAT4（-） group compared with those of STAT4（＋） group and IL-10 group. The splenic Foxp3 mRNA level was higher in STAT4（-） group than those of other three groups. Conclusion： After the therapy of supernatant of EIA cells with STAT4 negative expression, survival rate was improved and ascites were restrained significantly. However, the level of Foxp3 mRNA was significantly higher in spleen cells, which was possible to have detrimental effects on anti-tumor immunity in mice beating cancer.

关 键 词：STAT4转录因子 白细胞介素-10癌 Ehrlich瘤 存活率 DNA结合蛋白质类 

分 类 号：R457[医药卫生—治疗学] R657.3[医药卫生—临床医学]