干扰素α 2b通过降低MDSC抑制小鼠肾癌生长的研究

发布时间：2018-04-16 22:37

本文选题：外泌体 + MDSC　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:研究小鼠肾癌生长被干扰素α2b抑制及其潜在的作用机制。方法:1.建立肾癌移植瘤模型,将肾癌移植瘤模型分为对照组和干扰素α2b组,分别给予空白处理和干扰素α2b干预处理。建模后两组荷瘤小鼠移植瘤体积与重量进行观察监测,切取移植瘤作HE染色。2.外泌体通过超滤蔗糖密度梯度离心法提取,其形态及数量的变化通过透射电镜鉴定。3.在干扰素α2b组与对照组中,淋巴细胞应用FCM检测其所占百分比。4.在干扰素α2b组与对照组中,髓源性抑制性细胞应用FCM检测其所占百分比。5.BALB/c小鼠实验模型成功创建,将其分为生理盐水组、外泌体组和Renca细胞组,其后每隔10天荷瘤小鼠骨髓中骨髓源性抑制性细胞比例应用FCM检测。结果:1.肾癌移植瘤模型与BALB/c小鼠实验模型成功创建。在肾癌移植瘤模型实验中,与对照组相比,移植瘤的体积和重量经过干扰素α2b地处理后明显被抑制,HE染色检测结果显示:细胞核与细胞质比例失常,细胞核明显增大,细胞核染色加深,核分裂象明显异常,肿瘤组织中细胞无序,大小不一致,形态不均匀。2.成功分离并纯化外泌体,在对照组与干扰素组中,外泌体的形态并无明显差异;与对照组相比,干扰素α2b能使Renca细胞分泌的exosome数量明显减少。3.在干扰素α2b组中,MDSC与对照组比较,其所占百分比数明显下降。4.在对照组中,CD4/CD8与干扰素α2b组相比,其所占百分比数明显升高。5.在BALB/c小鼠实验模型实验中,与生理盐水组相比,外泌体组和Renca细胞组的荷瘤小鼠骨髓中的骨髓源性抑制性细胞百分比明显升高。外泌体组与Renca细胞组相比,荷瘤小鼠骨髓中的骨髓源性抑制性细胞比例无明显差异。外泌体组与Renca细胞组内,与前10天相比,后20天荷瘤小鼠骨髓中的骨髓源性抑制性细胞比例明显升高(P0.05),后20天之间荷瘤小鼠骨髓中的骨髓源性抑制性细胞比例无明显变化(P0.05)。NS组其百分比无明显变化(P0.05)。结论:Renca细胞来源的外泌体在小鼠体内参与了骨髓源性抑制性细胞的激活、募集,renca细胞分泌外泌体被干扰素α2b抑制,从而致使MDSCs的活化、增殖受限,使得移植肿瘤的发展过程明显受限。
[Abstract]:Aim: to study the inhibitory effect of interferon-伪-2 b on the growth of renal cell carcinoma in mice.Method 1: 1.The transplanted tumor model of renal cell carcinoma was established and divided into two groups: control group and interferon 伪 2b group, which were treated with blank treatment and interferon 伪 2b intervention respectively.After modeling, the volume and weight of transplanted tumor in two groups of mice were observed and monitored, and the transplanted tumor was removed for HE staining. 2.The exocrine was extracted by ultrafiltration sucrose density gradient centrifugation. The morphological and quantitative changes of exocrine were identified by transmission electron microscope (TEM).In interferon 伪 2b group and control group, lymphocytes were detected by FCM. 4. 4.In the interferon 伪 2b group and the control group, the percentage of myelogenous inhibitory cells was detected by FCM. 5. The BALB / c mouse model was successfully established and divided into normal saline group, exocrine group and Renca cell group.The proportion of bone marrow derived inhibitory cells in the bone marrow of tumor-bearing mice was detected by FCM every 10 days.The result is 1: 1.The transplanted tumor model of renal cell carcinoma and the experimental model of BALB/c mice were successfully established.In the model experiment of transplanted tumor of renal cell carcinoma, compared with the control group, the volume and weight of transplanted tumor were obviously inhibited by HE staining after treatment with interferon 伪 2b. The results showed that the ratio of nucleus to cytoplasm was abnormal and the nucleus increased obviously.The nuclear staining deepened, the mitotic image was obviously abnormal, the cells in the tumor tissue were disordered, the size was not consistent, and the morphology was not uniform. 2.The exocrine was isolated and purified successfully. There was no significant difference in the morphology of exocrine between the control group and the interferon group, and interferon 伪 2b could significantly reduce the number of exosome secreted by Renca cells.The percentage of MDSC in interferon 伪 2b group was significantly lower than that in control group.The percentage of CD4 / CD8 in the control group was significantly higher than that in the interferon 伪 2b group.The percentage of bone marrow-derived inhibitory cells in the bone marrow of tumor bearing mice in the exocrine group and Renca cell group was significantly higher than that in the saline group.There was no significant difference in the proportion of bone marrow-derived inhibitory cells between exocrine group and Renca cell group.In the exocrine group and Renca cell group, compared with the first 10 days,The proportion of bone marrow derived inhibitory cells in the bone marrow of tumor-bearing mice increased significantly after 20 days, while the percentage of bone marrow-derived inhibitory cells in bone marrow of tumor-bearing mice did not change significantly during the latter 20 days. There was no significant change in the percentage of bone marrow-derived inhibitory cells in the P0.05- NS group.Conclusion the exocrine derived from Renca cells is involved in the activation of bone marrow-derived inhibitory cells in mice, and the exocrine secreted by the recruitment of Renca cells is inhibited by interferon 伪 2b, which results in the activation and proliferation of MDSCs is restricted.The development of transplanted tumor is obviously limited.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.11

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