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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

28卷

期数:

2022年03期

页码:

253-259

栏目:

基础医学

出版日期:

2022-05-20

文章信息/Info

Title:

Construction and application of myeloid-specific SOCS3 gene knockout mice

文章编号:

1006-8147(2022)03-0253-07

作者:

纪辰燕; 李星晨; 陈桂冬; 于津浦

（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）

Author(s):

JI Chen-yan; LI Xing-chen; CHEN Gui-dong; YU Jin-pu

（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）

关键词:

Cre-loxP 系统; SOCS3 基因; 组织特异性; 小鼠; eMDSCs

Keywords:

Cre-loxP system; SOCS3 gene; tissue specificity; mouse; eMDSCs

分类号:

R332

DOI:

-

文献标志码:

A

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。

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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相似文献/References:

备注/Memo

备注/Memo:

基金项目国家自然科学基金面上项目（81872143）
作者简介:纪辰燕()1994-))，女，硕士在读，研究方向:)肿瘤学；
通信作 者：于津浦，E-mail：jyu@tmu.edu.cn。

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更新日期/Last Update: 2022-06-01