Preliminary study on the role of TM9SF2 knockdown in promoting the activity of the type I interferon signaling pathway to inhibit vesicular stomatitis virus replication.
- Author:
Kang LI
1
;
Xinyu WANG
2
;
Ran YE
2
;
Lingyun GUO
2
;
Linxu WANG
2
;
Nuo XU
3
;
Tong ZHANG
3
;
Xiaotao DUAN
4
Author Information
1. School of Pharmacy, Qingdao University, Qingdao 266071; National Key Laboratory of Special Drugs for National Security, Academy of Military Medical, Beijing 100850, China.
2. National Key Laboratory of Special Drugs for National Security, Academy of Military Medical, Beijing 100850, China.
3. Department of Stomatology, the First Medical Center, Chinese PLA General Hospital, Beijing 100853, China.
4. National Key Laboratory of Special Drugs for National Security, Academy of Military Medical, Beijing 100850, China. *Corresponding author, E-mail: xduan@ncba.ac.cn.
- Publication Type:Journal Article
- MeSH:
Humans;
Virus Replication/genetics*;
Signal Transduction;
Membrane Proteins/metabolism*;
A549 Cells;
Vesiculovirus/physiology*;
Interferon-beta/metabolism*;
Interferon Regulatory Factor-3/genetics*;
Interferon Type I/metabolism*;
Vesicular Stomatitis/immunology*;
Gene Knockdown Techniques;
Vesicular stomatitis Indiana virus/physiology*;
RNA, Small Interfering/genetics*
- From:
Chinese Journal of Cellular and Molecular Immunology
2025;41(6):481-487
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of the knockdown of transmembrane 9 superfamily protein member 2 (TM9SF2) on the replication of vesicular stomatitis virus (VSV), and investigate its role in the mechanism of antiviral innate immunity. Methods Small interfering RNA (siRNA) was used to knock down the TM9SF2 gene in human non-small cell lung cancer A549 cells. The CCK-8 method was used to assess cell proliferation. A VSV-green fluorescent protein (VSV-GFP) infected cell model was established. The plaque assay was used to measure the viral titer in the supernatant. RT-qPCR and Western blotting were employed to quantify the mRNA and protein levels of VSV genome replication in A549 cells following VSV infection, as well as the expression of interferon β (IFN-β) mRNA and interferon regulatory factor 3 (IRF3) protein phosphorylation following polyinosinic-polycytidylic acid (poly(I:C)) stimulation. Results Compared to the negative control, the knockdown of TM9SF2 exhibited a significant effect, with no observed impact on A549 cell proliferation. The VSV-GFP infected A549 cell model was successfully established. After viral stimulation, fluorescence intensity was reduced following TM9SF2 knockdown, and the mRNA and protein levels of VSV were significantly downregulated. The viral titer of VSV was decreased. After poly(I:C) stimulation, TM9SF2 knockdown significantly upregulated the mRNA level of IFN-β and the phosphorylation level of IRF3 protein. Conclusion The knockdown of TM9SF2 inhibits the replication of vesicular stomatitis virus, and positively regulates the type I interferon signaling pathway, thus enhancing the host's antiviral innate immune response.
