放疗联合抑制STAT3表达对恶性B细胞免疫原性分子表达的影响The Effect of Combination of Radiotherapy with Abrogating STAT3 Activity on the Maturation of Malignant B-cell Lymphoma Cells
张启芳,禹文峰,吴昌学,官志忠,柏华
ZHANG Qifang,YU Wenfeng,WU Changxue,GUAN Zhizhong,BAI Hua
摘要(Abstract):
目的:研究放疗联合抑制信号转导与转录激活因子3(STAT3)表达对恶性B细胞免疫原性的影响。方法:在BALB/c小鼠大腿皮下接种小鼠恶性B细胞淋巴瘤表达GFP的A20细胞形成肿瘤,在肿瘤内注射STAT3基因特异的shRNA质粒并进行放疗;采用荧光定量聚合酶链式反应(q PCR)和免疫印迹法(Western blot)方法分别检测肿瘤组织中STAT3 mRNA和蛋白表达水平;细胞膜蛋白分子染色法检测恶性B细胞表面分子MHCII、CD40和CD80的表达,用流式细胞仪收集细胞染色数据。结果:放疗联合抑制STAT3表达能显著降低STAT3 mRNA和蛋白表达水平,与单纯放疗比较,放疗联合抑制STAT3表达治疗显著增加A20细胞表面分子MHCII、CD40及CD80的表达水平,差异有统计学意义(P<0.05)。结论:放疗联合抑制STAT3表达能增强决定恶性B细胞免疫原性的分子表达。
Objective: To examine the effect of combination of radiotherapy with STAT3 inhibition on the immunogenicity of malignant B-cell Lymphoma Cells. Methods: Murine A20 B cell lymphoma cells were subcutaneously injected the thighs of BALB / c mice to form tumors. Tumors were given intratumor injection of STAT3 shRNA plasmid and local radiotherapy. STAT3 expression in tumors was determined by q PCR at mRNA levels and by Western blot at protein levels. The expression of MHCII,CD40 and CD80 were determined by extracellular staining and subsequent data collected by flow cytometry. Results: STAT3 expressions were remarkably silenced in tumors. Compared to radiotherapy alone,radiotherapy combined with STAT3 silencing significantly upregulated the expression of MHCII,CD40 and CD80 on A20 cells. Conclusions: Radiotherapy plus STAT3 silencing increases the expression of molecules that decide the immunogenicity of malignant B cell lymphoma.
关键词(KeyWords):
信号转导与转录激活因子3;放射疗法;淋巴瘤,B细胞;聚合酶链式反应;免疫印迹;流式细胞术
signal transducer and activator of transcription 3;radiotherapy;lymphoma,B cell;polymerase chain reaction;western blot;flow cytometry
基金项目(Foundation): 国家自然科学基金项目(No.81560482);; 贵州省“2011协同创新中心”支助项目[黔教合协同创新中心(2014)06];; 贵州省科技厅重大专项[黔科合计Z字(2012)4010];; 贵州省科技厅科技计划项目[黔科合LG字(2012)009]
作者(Author):
张启芳,禹文峰,吴昌学,官志忠,柏华
ZHANG Qifang,YU Wenfeng,WU Changxue,GUAN Zhizhong,BAI Hua
DOI: 10.19367/j.cnki.1000-2707.2015.11.006
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- 信号转导与转录激活因子3
- 放射疗法
- 淋巴瘤,B细胞
- 聚合酶链式反应
- 免疫印迹
- 流式细胞术
signal transducer and activator of transcription 3 - radiotherapy
- lymphoma,B cell
- polymerase chain reaction
- western blot
- flow cytometry