Cbl-b基因介导T细胞免疫杀伤小鼠LA795肺腺癌细胞的实验研究  

The experimental research about immunity killing of Cbl-b gene mediated T lymphocyte against Mouse LA795 lung neoplasms cells

作　　者：黄海涛[1] 张华[2] HUANG Hai-tao ZHANG Hua(Department of Respiratory Medicine, the Fifth People＇s Hospital of Chengdu, Chengdu, Sichuan 611130, China)

机构地区：[1]成都市第五人民医院呼吸内科,四川成都611130 [2]成都市第五人民医院消化内科,四川成都611130

出　　处：《临床肺科杂志》2017年第3期538-541,共4页Journal of Clinical Pulmonary Medicine

摘　　要：目的利用特异性小干扰RNA(small interfering RNA,siRNA)沉默T细胞Cbl-b基因表达,观察转染T细胞对小鼠肺腺癌细胞LA795的体外免疫杀伤作用。方法筛选高效特异性沉默Cbl-b基因的siRNA序列转染T739小鼠脾脏T细胞,转染72h后,利用酶联免疫吸附法(ELISA)检测IL-2、INF-γ等T细胞免疫因子分泌情况,比较单纯T细胞、阴性对照T细胞及转染T细胞与小鼠肺腺癌细胞LA795混合培养肿瘤杀伤率。结果转染72h后,转染组细胞因子IL-2、INF-γ分泌水平较空转组和空白组显著增加。在体外实验中,与单纯T细胞及阴性对照T细胞相比,转染T细胞能更高效杀伤小鼠肺腺癌细胞LA795,最高杀瘤率达到58.38±3.82%。结论利用特异性siRNA技术沉默Cbl-b基因能够促进小鼠T细胞因子IL-2和INF-γ分泌,增强T细胞对肺腺癌细胞LA795的体外免疫杀伤作用。Objective To investigate the cytotoxicity activity of transfected T lymphocytes against LA795 lung neoplasms cells in vitro by utilizing specific small interfering RNA （siRRA） to silence the expression of casitas B cell lymphoma-b（Cbl-b） gene of T-Lymphocytes. Methods T lymphocytes were transfected by specific siRNA to silence the expression of casitas B cell lymphoma-b （Cbl-b） gene. 72 hours after transfection, the secretion of IL-2 and IFN-γ were measured by ELISA. In the end, the cytotoxicity activity changes against LA795 lung neoplasms cells were compared among transfected T lymphocytes, negative control and blank control in vitro. Results 72 hours after transfection, compared with the other groups, T lymphocytes transfected with siRNA showed higher secretion level of IL-2 and IFN-γ （P 〈0. 05）. Transfected T lymphocyte also showed more efficient killing ability against LA795 lung neoplasms cells than negative control and blank control in vitro. The highest killing rate reached 58.38 ± 3.82%. Conclusion Silencing Cbl-b can significantly promote the secretion level of IL-2 and IFN-γ of mice T lymphocyte, and enhance the cytotoxicity activity of T lymphocyte against LA795 lung neoplasms ceils in vitro.

关 键 词：CBL-B 基因沉默 T淋巴细胞 肺腺癌 免疫疗法 

分 类 号：R734.2[医药卫生—肿瘤]