摘要
目的:研究干扰素α2b抑制小鼠肾癌生长及其潜在的作用机制。方法:建立肾癌移植瘤模型,干扰素干预后将肾癌移植瘤模型分为对照组和干扰素α2b组,监测各组小鼠移植瘤体积与重量,切取移植瘤作HE染色,免疫组织化学检测移植瘤蛋白BCL-2表达;超滤加蔗糖重水密度梯度离心法提取外泌体,透射电镜鉴定外泌体形态及数量的变化;流式细胞术检测小鼠骨髓中髓源性抑制细胞(myeloid-derived suppressor cells,MDSCs)比例,小鼠脾脏CD4/CD8T淋巴细胞比例。建立BALB/c小鼠实验模型,将其分为生理盐水组、外泌体组和Renca细胞组,建模后第10、20、30天应用流式细胞仪检测各组小鼠骨髓MDSCs比例。结果:成功建立肾癌移植瘤模型与BALB/c小鼠实验模型,在肾癌移植瘤模型实验中,与对照组比较,干扰素α2b组重量与体积明显被抑制(P<0.01)。HE染色结果显示:肿瘤组织中细胞大小不均,形态不一,排列紊乱;细胞核增大,核质比例失常,核深染,异常核分裂。免疫组织化学检测显示:对照组为、干扰素α2b组两组间Bcl-2蛋白阳性表达率无统计学差异(P>0.05)。成功分离并纯化外泌体,对照组、干扰素组2组外泌体形态并无统计学差异(P>0.05);与对照组比较,干扰素α2b能明显减少Renca细胞分泌的外泌体数量(P<0.01)。与对照组比较,干扰素α2b组MDSCs比例明显降低(P<0.05),与对照组相比,干扰素组荷瘤小鼠CD4/CD8T淋巴细胞比例(P<0.05)明显升高。在BALB/c小鼠实验模型实验中,在第10天、20天和30天,与生理盐水组相比,外泌体组与Renca细胞组的MDSCs比例明显增高(P<0.01),而外泌体组与Renca细胞组比较无统计学差异(P>0.05)。外泌体组与Renca细胞组内,第20天和30天比第10天要明显增加MDSCs比例(P<0.05),第20天和30天之间MDSCs比例无明显变化(P>0.05)。而生理盐水组MDSCs比例在第10、20和30天之间比较无统计学差异(P>0.05)。结论:干扰素α2b通过抑制R
Objective：to investigate the effect of interferon α2b on the growth of renal carcinoma in mice,and to explore the potential mechanism. Methods：Renal carcinoma xenograft model was established. After intervention of interferon,transplanted renal carcinoma model was divided into control group and interferon α2b group. The volume and weight of transplanted tumor in mice was monitored,and tumor was removed was for HE staining. BCL-2 expression was detected in transplanted tumor by immunohistochemical method.Exosomes from renca cells were separated and purified by ultrafiltration and sucrose gradient centrifugation. The morphology and quantitative of exosomes was identified by transmission electron microscope. The proportion of MDSCs in bone marrow and spleen CD4/CD8 T lymphocytes of mice was determined by flow cytometry. BALB/c mouse model was established and was divided into normal saline group,exosomes group and renca cell group. On 10,20,30 days after modeling,the MDSCs ratio of bone marrow in each group was detected by flow cytometry. Results：The tumor model of renal carcinoma and experimental model of BALB/c mice were successfully established. Compared with those of control model,the volume and weight of the transplanted tumor treated with interferon α2b were significantly inhibited（P〈0.01）. HE staining showed that the tumor cells varied in size and morphology,arranged in disorder and had increased nucleus,nucleo cytoplasmic ratio arrhythmia,hyperchromatic nuclei,and abnormal nuclear fission. Immunohistochemistry showed that there was no significant difference in the expression of Bcl-2 protein between the two groups（P〈0.05）. The exosome was separated and purified. There was no significant difference between the two groups in exosomes（P〈0.05）. And the interferon α2b could significantly reduce the amount of exosomes secreted by Renca cells compared with that of control group（P=0.000）. Compared with that of control group,the proportion of MDSCs in interferon α2b group was significan
作者
殷永健
陈仕友
罗春丽
张尧
吴小候
Yin Yongjian Chen Shiyou Luo Chunli Zhang Yao Wu Xiaohou(Department of Urology, The First Affiliated Hospital of Chongqing Medical University College of Laboratory Medicine ,Chongqing Medical University)
出处
《重庆医科大学学报》
CSCD
北大核心
2017年第5期473-480,共8页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:81272572)
重庆市卫生局科研资助项目(编号:2012242)
关键词
干扰素Α2B
外泌体
髓源性抑制细胞
肾癌
抑制生长
interferon α2b
exosomes
myeloid-derived suppressor cells
renal carcinoma
inhibiting growth
