瘤体注射Cbl—b基因真核表达质粒转染T淋巴细胞免疫治疗小鼠前列腺癌  

Immunotherapy mouse prostate cancer with direct intratumorai injection of casitas B cell lympho- ma-b gene Eukaryotic vectors transfection of T lymphocytes

作　　者：王顺平[1] 周术奎[1] 史振铎[1] 陈卫华[1] 李良[1] 王跃闽[1] 

机构地区：[1]同济大学附属东方医院泌尿外科,上海200120

出　　处：《中华实验外科杂志》2013年第6期1167-1170,共4页Chinese Journal of Experimental Surgery

基　　金：基金项目：上海科委基金资助项目（PKJ2008-Y43）

摘　　要：目的瘤体注射Cbl—b基因真核表达质粒[短发卡RNA（shRNA）一Cbl—b]转染T淋巴细胞，观察前列腺癌小鼠肿瘤生长及机体免疫水平变化。方法尼龙毛柱法提取小鼠脾脏T淋巴细胞，质粒转染shRNA-Cbl—b至T淋巴细胞；瘤体注射shRNA—Cbl—bT淋巴细胞至RM-1前列腺癌小鼠肿瘤部位，每周1次，共4次；Westernblot法检测荷瘤小鼠肿瘤组织中Cbl—b蛋白的表达，测量小鼠肿瘤体积变化；酶联免疫吸附试验（ELISA）法检测小鼠外周血中的白细胞介素（IL-2）、吖干扰素（IFN-1）、肿瘤坏死因子-β（TNF-β）水平变化。结果与阴性对照组和空白组比较，瘤体注射shRNA—Cbl-bT细胞能降低肿瘤组织Cbl-b蛋白表达，显著抑制小鼠前列腺肿瘤生长（P〈0．05）；shRNA—Cbl—b组、阴性对照组、空白组外周血中IL-2浓度分别为（560．34±21．29）、（447．05±20．11）、（445．12±15．06）ng／L（P〈0．05），IFN-1浓度分别为（435．70±22．25）、（403．37±7．15）、（380．02±14．43）ng／L（P〈0．05），TNF—β浓度分别为（306．81±9．71）、（253．15±9．34）、（253．38±15．67）ng／L（P〈0．05）。结论瘤体注射shRNA-Cbl—b T淋巴细胞能抑制小鼠前列腺肿瘤的生长，提高外周血IL-2、IFN-γ、TNF—β等细胞因子分泌水平，增强了小鼠机体抗前列腺肿瘤免疫。Objective To investigate the mouse prostate cancer growth and the immune level change, after direct intratumoral injection of casitas B cell lymphoma-b （Cbl-b） eukaryotic expression plas- mid short hairpin RNA [ shRNA） -Cblb] transfection of T lymphocytes. Methods T lymphocytes were i- solated from the spleen of C57BL/6 mice by nylon wool fiber columns, and then were transfeced Cbl-b gene by Eukaryotie vectors ; direct intratumoral injection of shRNA-Cbl-b T lymphoeytes was done once a week, and for a total of four times; Western blotting method were used to examine Cbl-b protein expression in mice tumor; changes of tumor volume in mice were monitored; Enzyme linked immunosorbent assay （ELISA） were used to examine peripheral blood interferon （IFN）-7, interleukin （IL）-2 and tumor necro- sis factor （TNF） -β production in tumor-bearing mice. Results After injection the transfeetion of T lym- phocytes into mites, compared with the control group and the blank group, shRNA-Cbl-b T cells can re- duce tumor Cbl-b protein expression and significantly inhibit prostate tumor growth （P 〈 0. 05 ）. IL-2 con- centration of peripheral blood in shRNA-Cbl-b group, negative control group, blank group were （560. 34±21.29）, （447. 05±20. 11 ）, （445.12±15.06） ng/L （P 〈0. 05）. IFN-7 concentration of peripheral blood in shRNA-Cbl-b group, negative control group, blank groupwere （435.70±22. 25 ）, （403. 37± 7. 15 ）, （380. 02±14. 43） ng/L （P 〈0. 05）. TNF-β concentration of peripheral blood in shRNA-Cbl -b group, neg-ative control group, blank group were （306. 81±9. 71 ）, （253.15 ±9. 34 ）, （253.38 ±15. 67 ） ng/L （ P 〈 0. 05 ）. Conclusion The mouse prostate cancer growth can be significantly inhabited by direct intratumoral injection of shRNA-Cbl-b transfection of Y lymphocytes, and IL-2, IFN-γ, TNF-β and other cytokines secretion levelof peripheral blood increase largely, All of these increase the effect of prostate cancer immune in mice.

关 键 词：CBL-B 短发卡RNA 基因沉默 T淋巴细胞 前列腺癌 肿瘤免疫 

分 类 号：R737[医药卫生—肿瘤]