| 髓系特异性SOCS3 基因敲除小鼠的构建及应用 |
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| 引用本文： | 纪辰燕,李星晨,陈桂冬,于津浦.髓系特异性SOCS3 基因敲除小鼠的构建及应用[J].天津医科大学学报,2022,0(3):253-259. |
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| 作者姓名： | 纪辰燕 李星晨 陈桂冬 于津浦 |
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| 作者单位： | （天津医科大学肿瘤医院肿瘤分子诊断中心，国家肿瘤临床医学研究中心，天津市“肿瘤防治”重点实验室，
天津恶性肿瘤临床医学研究中心，天津市肿瘤免疫与生物治疗重点实验室，天津300060） |
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| 摘 要： | 目的:通过Cre-loxP 基因敲除系统特异性敲除髓系SOCS3 基因，构建早期髓系来源抑制细胞（eMDSCs）高浸润荷瘤鼠模
型。方法: 通过将SOCS3fl/-小鼠与Lyz2-Cre 小鼠杂交繁育， 获得髓系特异性SOCS3 基因敲除小鼠，PCR 法鉴定小鼠基因型，
Western 印迹验证基因敲除效果， 流式细胞术检测髓系特异性SOCS3 基因敲除小鼠骨髓中eMDSCs 比例及其对T 细胞的抑制
作用， 并在该小鼠基础上分别构建乳腺癌、肺癌和黑色素瘤3 种eMDSCs 高浸润荷瘤鼠模型， 流式细胞术检测肿瘤组织中
eMDSCs 浸润情况。结果:PCR 鉴定和Western印迹检测证实髓系特异性SOCS3 基因敲除小鼠构建成功，流式细胞术结果表明
髓系SOCS3 基因敲除小鼠骨髓中eMDSCs 比例显著升高（t=17.94，P＜0.001），且该群eMDSCs 抑制T 细胞增殖（t=14.21，
P＜0.001）、促进T 细胞凋亡（t=13.53，P＜0.001）。在黑色素瘤、乳腺癌、肺癌3 种髓系特异性SOCS3 基因敲除荷瘤鼠的肿瘤组织
中eMDSCs 数量显著增加（t=24.14、24.56、14.93，均P＜0.001）。结论:通过髓系特异性SOCS3敲除可成功构建eMDSCs 高浸润荷
瘤鼠模型。
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| 关 键 词： | Cre-loxP 系统 SOCS3 基因 组织特异性 小鼠 eMDSCs |
Construction and application of myeloid-specific SOCS3 gene knockout mice |
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| JI Chen-yan,LI Xing-chen,CHEN Gui-dong,YU Jin-pu.Construction and application of myeloid-specific SOCS3 gene knockout mice[J].Journal of Tianjin Medical University,2022,0(3):253-259. |
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| Authors: | JI Chen-yan LI Xing-chen CHEN Gui-dong YU Jin-pu |
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| Affiliation: | （Cancer Molecular Diagnostics Core，Tianjin Medical University Cancer Institute and Hospital，National Clinical Research Center of
Cancer，Key Laboratory of Cancer Prevention and Therapy，Tianjin′ s Clinical Research Center for Cancer，Key Laboratory of Cancer
Immunology and Biotherapy，Tianjin 300060，China） |
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| Abstract: | Objective:To construct mouse models of cancer with high infiltration of eMDSCs through myeloid-specific SOCS3 gene
knockout mice based on the Cre-loxP system. Methods: SOCS3fl/fl mice were crossed with Lyz2-Cre mice to generate myeloid-specific
SOCS3 gene knockout mice.The genotypes of the offspring were identified by PCR，and the knockout effect was verified by Western
blotting.The proportion of eMDSCs in the bone marrow of myeloid-specific SOCS3 gene knockout mice and their inhibitory effection T cells
were detected by flow cytometry. Based on the myeloid-specific SOCS3 gene knockout mice，three kinds of tumor-bearing mouse model s
include breast cancer，lung cancer，and melanoma with high infiltration of eMDSCs were constructed，and the infiltration of eMDSCs in
tumors was detected by flow cytometry. Results: PCR results and Western blotting analysis confirmed that myeloid-specific SOCS3 gene
knockout mice were successfully constructed. Flow cytometry results demonstrated that the proportion of eMDSCs in the bone marrow of
myeloid-specific SOCS3 gene knockout mice was significantly increased（t=17.94，P＜0.001），and the eMDSCs significantly inhibited T cell
proliferation（t=14.21，P＜0.001）and promoted T cell apoptosis（t=13.53，P＜0.001）.The number of eMDSCs was significantly increased in
tumor tissue of three types of myeloid-specific SOCS3 gene knockout tumor-bearing mice with melanoma，breast cancer，and lung cancer
（t=24.14，24.56，14.93，all P＜0.001）. Conclusion: Mouse models of cancer with high infiltration of eMDSCs are successfully constructed
by specifically knocking out the SOCS3 gene of mouse myeloid cells. |
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| Keywords: | Cre-loxP system SOCS3 gene tissue specificity mouse eMDSCs |
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