Basic & Clinical Medicine ›› 2023, Vol. 43 ›› Issue (1): 130-136.doi: 10.16352/j.issn.1001-6325.2023.01.0130

• Original Articles • Previous Articles     Next Articles

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

MENG Xuanyu1#, LIU Dihui2#, JIANG Shuo2, GUAN Yaqun2*, LIANG Xiaodi2*   

  1. 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

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

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