敲低Stau1后小鼠前脂肪细胞系3T3-L1中FOXP1的表达增加

doi:10.16352/j.issn.1001-6325.2023.01.0130

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (1): 130-136.doi: 10.16352/j.issn.1001-6325.2023.01.0130

敲低Stau1后小鼠前脂肪细胞系3T3-L1中FOXP1的表达增加

孟轩羽^1#, 刘迪晖^2#, 蒋硕², 关亚群^2*, 梁小弟^2*

新疆医科大学基础医学院 1.机能中心; 2.生物化学与分子生物学教研室,新疆维吾尔自治区乌鲁木齐 830017

收稿日期:2021-09-20 修回日期:2022-06-02 发布日期:2022-12-27
通讯作者: ^*yaqunguan557@xjmu.edu.cn;xiaodiliang@xjmu.edu.cn
^#对本文有相同贡献
基金资助:
新疆维吾尔自治区2017年度高校科研计划(XJEDU2017M016);省部共建中亚高发病成因与防治国家重点实验室代谢病专项(SKL-HIDCA-2021-DX5);新疆维吾尔自治区研究生科研创新项目(XJ2021G223)

FOXP1 expression is increased after Stau1 knockdown of murine preadipocyte cell line 3T3-L1

MENG Xuanyu^1#, LIU Dihui^2#, JIANG Shuo², GUAN Yaqun^2*, LIANG Xiaodi^2*

1. Functional Center;
2. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xinjiang Medical University, Urumqi 830017, China

Received:2021-09-20 Revised:2022-06-02 Published:2022-12-27
Contact: ^*yaqunguan557@xjmu.edu.cn;xiaodiliang@xjmu.edu.cn

摘要/Abstract

摘要： 目的在小鼠前脂肪细胞系3T3-L1分化过程中敲低双链RNA结合蛋白Staufen1(Stau1)后筛选差异表达的基因,分析其生物学功能,并用qPCR验证测序结果。方法用RNA-seq分析Stau1敲低后基因的转录调控。构建STAU1 shRNA并转染3T3-L1细胞,鸡尾酒方法诱导其分化为成熟脂肪细胞,收取0、4 d的细胞设立对照组和STAU1敲低组(各3组生物重复样本)收取12组样品的细胞基因芯片信息,以变化倍数log2(Fold change)＞1且校正后的P<0.05作为条件筛选Stau1敲低的细胞与对照组样本间的差异表达基因,进行基因本体论(GO)及代谢通路分析(KEGG通路数据库),qPCR验证差异表达的基因。荧光素酶报告基因实验验证SATU1与叉头盒蛋白P1 FOXP1 mRNA 3′UTR上Staufen1结合位点(SBS)存在结合。结果较对照组STAU1敲低细胞组中共筛选出差异表达基因588个,其中上调406个、下调182个。差异表达基因主要参与的生物学过程中脂代谢、炎性反应因子,主要富集的信号通路有糖代谢和能量代谢相关通路。敲低Stau1后FOXP1表达升高5.3倍,软件预测FOXP1 mRNA 3′UTR含有STAU1结合位点,荧光素酶报告基因验证了FOXP1 mRNA 3′UTR含有STAU1结合位点。结论推测STAU1能够与FOXP1 mRNA上的STAU1结合位点结合并促进其降解。

关键词: Staufen1, 叉头盒蛋白p1, 转录组测序, 脂肪细胞分化

Abstract: Objective To screen the differentially expressed genes after knocking down double-stranded RNA binding protein stanfen 1(Stau1)during the differentiation process of murine preadipocyte cell line 3T3-L1, analyze their biological functions, and use qPCR to verify the sequencing results for analysis. Methods RNA-seq was used to analyze the transcriptional regulation of genes after Stau1 knockdown. The STAU1 shRNA was constructed and transfected into 3T3-L1 cells. The cocktail method induced them to differentiate into mature adipocytes. The cells of 0 and 4 days were collected to establish a control cell group and a STAU1 knockdown group (3 groups of biological replicate samples) for a total of 12 groups of samples. Cell gene chip data set, with change log2 (Fold change)＞ 1 and corrected P<0.05 as the screening criteria to screen the differentially expressed genes between Stau1 knockdown cells and control group samples, and then the fat after knockdown Gene ontology (GO) and metabolic pathway analysis (KEGG pathway database) were performed on the differentially expressed genes of cells and control groups. Luciferase experiments confirmed the presence of SBS binding to forkhead box P1(FOXP1) mRNA 3′UTR. Results Compared with the control, STAU1 knockdown cell group, a total of 588 differentially expressed genes were screened, of which 406 were up-regulated and 182 were down-regulated. Differentially expressed genes were mainly involved in lipid metabolism and inflammatory responded factors in the biological process, and the main enriched signal pathways were related to sugar metabolism and energy metabolism. FOXP1 expression increased 5.3 times after knocking down Stau1, and the software predicted that the 3′UTR region of FOXP1 mRNA contained STAU1 binding sites. Conclusions Therefore, it is speculated that STAU1 can bind to the STAU1 binding site loacated at FOXP1 mRNA and promote its degradation.

Key words: staufen1, forkhead box p1, transcriptomic sequencing, adipocyte differentiation

中图分类号:

孟轩羽, 刘迪晖, 蒋硕, 关亚群, 梁小弟. 敲低Stau1后小鼠前脂肪细胞系3T3-L1中FOXP1的表达增加[J]. 基础医学与临床, 2023, 43(1): 130-136.

MENG Xuanyu, LIU Dihui, JIANG Shuo, GUAN Yaqun, LIANG Xiaodi. FOXP1 expression is increased after Stau1 knockdown of murine preadipocyte cell line 3T3-L1[J]. Basic & Clinical Medicine, 2023, 43(1): 130-136.

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敲低Stau1后小鼠前脂肪细胞系3T3-L1中FOXP1的表达增加

FOXP1 expression is increased after Stau1 knockdown of murine preadipocyte cell line 3T3-L1

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