[1]王 迪,王润杨,杨俊寒,等. 表达人源PD-L1的4T1细胞系的构建[J].湖北工业大学学报,2022,(5):76-78.
　WANG Di,WANG Runyang,YANG Junhan,et al. Construction of 4T1 Cell Line Expressing Human PD-L1[J].,2022,(5):76-78.
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 表达人源PD-L1的4T1细胞系的构建()
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《湖北工业大学学报》[ISSN:1003-4684/CN:42-1752/Z]

卷:
期数:
2022年第5期
页码:
76-78
栏目:
湖北工业大学学报
出版日期:
2022-10-25

文章信息/Info

Title:
 Construction of 4T1 Cell Line Expressing Human PD-L1
文章编号:
1003-4684(2022)05-0076-03
作者:
 王 迪;  王润杨;  杨俊寒;  张 乐;  胡 翰;  汪 洋;  刘滨磊
 湖北工业大学生物工程与食品学院， 湖北 武汉 430068
Author(s):
 WANG Di; WANG Runyang; YANG Junhan; ZANG Le; HU Han; WANG Yang; LIU Binlei
 School of Food and Biological Engin., Hubei Univ., of Tech .Wuhan 430068, China
关键词:
 ; 4T1细胞;  PD-L1;  绿色荧光蛋白
Keywords:
 ; 4T1;  PD-L1;  green fluorescent protein
分类号:
R153
文献标志码:
A
摘要:
 为构建稳定表达人源hPD-L1分子的小鼠乳腺癌细胞系（4T1），建立BALB/c小鼠乳腺癌肿瘤模型，采用Lipofactamine 3000转染试剂将携带绿色荧光蛋白（Green fluorescent protein,GFP）和hPD-L1的质粒转入4T1细胞中，经嘌呤霉素筛选获得4T1-hPD-L1单克隆细胞系，通过流式细胞术检测细胞纯度。将构建成功的细胞系皮下植入BALB/c小鼠背部，建立乳腺癌皮下肿瘤模型。流式细胞术检测4T1-hPD-L1细胞中GFP、PD-L1双阳性率为97.0%。将4T1-hPD-L1单克隆细胞系皮下接种于BALB/c小鼠背部可成瘤。成功构建稳定表达hPD-L1分子的4T1单克隆细胞系，并建立BALB/c小鼠乳腺癌肿瘤模型，为进一步研究PD-L1抗体药物提供参考。
Abstract:
 This paper is to construct mouse breast cancer cell line (4T1) stably expressing human PD L1 molecule, and to establish BALB/c mouse breast cancer tumor model. The plasmid carrying green fluorescent protein (GFP) and PD-L1 was transfected into 4T1 cells with Lipofactamine 3000 transfection reagent, and 4T1-hPD-L1 monoclonal cell lines were obtained by purinamycin screening. Cell purity was determined by flow cytometry. The successfully constructed cell lines were implanted subcutaneously into the back of BALB/c mice to establish the transplanted breast cancer tumor model. Results show that the double positive rate of GFP and PD-L1 in 4T1-hPD-L1 cell was 97.0% by flow cytometry. The 4T1-hPD-L1 monoclonal cell line was inoculated subcutaneously into the back of BALB/c mice to form tumors. The 4T1 monoclonal cell line with stable expression of human PD-L1 molecule was successfully constructed, and the breast cancer tumor model of BALB/c mice was established, which laid a foundation for further study of PD-L1 antibody drugs.

参考文献/References:

[1] 宋莉, 葛彦, 曹莎莎, 等. 人PD-L1基因转染细胞株的构建及其对活化的Jurkat细胞增殖和凋亡的影响[J]. 江苏大学学报, 2015,25(2):123-127.
[2] ZOU W, CHEN L. Inhibitory B7-family molecules in the tumour microenvironment[J]. Nature Reviews Immunology, 2008,8(6):467-477.
 [3] HIRANO F, KANEKO K, TAMURA H, et al. Blockade of B7-H1 and PD-1 by monoclonal antibodies potentiates cancer therapeutic immunity[J]. Cancer Research, 2005,65(3):1089-1096.
[4] 朱云霞, 胡振华, 陈永井, 等. 人PD-L1真核表达载体的构建及其在HEK293细胞系中的稳定转染[J]. 苏州大学学报, 2009,29(3):459-462.
[5] 张郧樊, 杨建新. PD-1/PD-L1信号通路及其在乳腺癌治疗中的研究进展[J]. 中国医学创新, 2017,14(2):145-148.
[6] 李佳, 敖宁, 乔东波, 等. PD-L1表达与乳腺癌临床病理特征及预后关联Meta分析[J]. 中华肿瘤防治杂志, 2018,25(19):1395-1402.
[7] 屠珏, 黄宇, 凌云, 等. 电转系统介导的荧光素酶稳定表达PANC-1细胞的构建及其生物发光成像检测[J]. 中国细胞生物学学报, 2016,38(7):37-38.
[8] 段海潇, 杨俊寒, 王琳, 等. 表达近红外荧光蛋白的结肠癌细胞系的建立[J]. 湖北工业大学学报, 2020,35(5):71-74.
[9] 刘向云, 杨荣富, 胡文娟, 等. 人OC-3-VGH卵巢癌细胞裸小鼠肿瘤模型的建立[J]. 中国实验动物学报, 2009,17(2):108-11.0
[10] 田季平, 张剑, 周金培, 等. 免疫检查点PD-1/PD-L1小分子抑制剂的研究进展[J]. 中国药科大学学报, 2019,50(1):1-10.
[11] OKAZAKI T, HONJO T. PD-1 and PD-1 Ligands:from discovery to clinical application[J]. International Immunology, 2007,19(7):813-824.
[12] 雷占翔, 刘芳芳, 于才勇, 等. PB-GF转座子系统的构建及其在真核细胞中转座的研究[J]. 细胞与分子免疫学杂志, 2014,30(6):652-655.

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备注/Memo

备注/Memo:
［收稿日期］ 2021-07-26
［第一作者］ 王 迪（1997-），女，安徽安庆人，湖北工业大学硕士研究生，研究方向为生物工程
［通信作者］ 刘滨磊（1962-），男，湖北武汉人，湖北工业大学教授，研究方向为生物制药
更新日期/Last Update: 2022-10-26