干扰素α2b通过降低MDSC抑制小鼠肾癌生长的研究

发布时间：2018-05-01 16:37

本文选题：干扰素αb + 外泌体　；参考：《重庆医科大学学报》2017年05期

【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(P0.01)。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(P0.05)。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(P0.05);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(P0.01)。与对照组比较,干扰素α2b组MDSCs比例明显降低(P0.05),与对照组相比,干扰素组荷瘤小鼠CD4/CD8T淋巴细胞比例(P0.05)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(P0.01),而外泌体组与Renca细胞组比较无统计学差异(P0.05)。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(P0.05),第20天和30天之间MDSCs比例无明显变化(P0.05)。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(P0.05)。结论:干扰素α2b通过抑制Renca细胞分泌外泌体,而Renca细胞来源外泌体在体内参与了小鼠MDSCs的活化、增殖,致使MDSCs的活化、增殖受限,从而抑制小鼠肾癌生长。
[Abstract]:Aim: to study the inhibitory effect of interferon-伪-2b on the growth of renal cell carcinoma in mice and its potential mechanism. Methods: the transplanted tumor model of renal cell carcinoma was established. After interferon intervention, the transplanted tumor model was divided into control group and interferon 伪 2b group. The volume and weight of transplanted tumor in each group were monitored, and the transplanted tumor was removed for HE staining. The expression of BCL-2 was detected by immunohistochemistry, the exocrine was extracted by ultrafiltration and sucrose heavy water density gradient centrifugation, and the morphological and quantitative changes of exocrine were identified by transmission electron microscope (TEM). The ratio of myeloid-derived suppressor cells in bone marrow and the proportion of CD4/CD8T lymphocytes in spleen of mice were detected by flow cytometry. BALB/c mice were divided into three groups: normal saline group, exocrine group and Renca cell group. Flow cytometry was used to detect the proportion of MDSCs in bone marrow of each group. Results: the transplanted tumor model of renal cell carcinoma and the experimental model of BALB/c mice were successfully established. Compared with the control group, the weight and volume of the interferon 伪 2b group were significantly inhibited by P0.01he staining. The results showed that the size of the cells in the tumor tissue was uneven. The nucleus is enlarged, the proportion of nucleus and cytoplasm is abnormal, the nucleus is deep stained and abnormal mitosis. Immunohistochemical examination showed that the positive expression rate of Bcl-2 protein in interferon 伪 2b group was not significantly different from that in interferon 伪 2b group (P 0.05). The exocrine was isolated and purified successfully. There was no significant difference in the morphology of exocrine in control group and interferon group (P 0.05). Compared with control group, interferon 伪 2b could significantly reduce the number of exocrine bodies secreted by Renca cells. Compared with the control group, the proportion of MDSCs in the interferon 伪 2b group was significantly lower than that in the control group. Compared with the control group, the proportion of CD4/CD8T lymphocytes in the interferon group was significantly higher than that in the control group. In the BALB/c mouse model experiment, on the 10th day, 20 days and 30 days, compared with the normal saline group, the proportion of MDSCs in the exocrine group and Renca cell group was significantly higher than that in the control group, but there was no significant difference between the exocrine body group and the Renca cell group. In the exocrine group and the Renca cell group, the proportion of MDSCs was significantly increased on the 20th and 30th days as compared with the 10th day, but there was no significant change in the MDSCs ratio between the 20th and the 30th day. However, there was no significant difference in the proportion of MDSCs between the 10 ~ (th) day and the 30 ~ (th) day in the saline group (P 0.05). Conclusion: interferon 伪 2b inhibits the exocrine secretion of Renca cells, and the exocrine derived from Renca cells participate in the activation and proliferation of mouse MDSCs in vivo, resulting in the limited activation and proliferation of MDSCs, thus inhibiting the growth of mouse renal cell carcinoma.
【作者单位】：重庆医科大学附属第一医院泌尿外科;重庆医科大学检验医学院;
【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)
【分类号】：R737.11

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