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Updated: Jul 2, 2026
Using RNA-interference to Investigate the Innate Immune Response in Mouse Macrophages
Published on: November 3, 2014
Circ_0035292 knockdown alleviates lipopolysaccharide (LPS)-induced WI-38 cell apoptosis and inflammatory injury
Ying Guo1, Zhouzhen Li1, Chen Cheng1
1Department of Pediatrics, Wuhan Asia General Hospital, Wuhan City, Hubei, China.
Background:
Circular RNAs have emerged as important regulators in the pathogenesis of human diseases, including infantile pneumonia (IP). In this study, we aimed to explore the effects of circ_0035292 on lipopolysaccharide (LPS)-treated Wistsar Institute (WI)-38 cells.
Methods:
Quantitative real-time polymerase chain reaction and western blot were executed to detect the levels of circ_0035292, microRNA-370-3p (miR-370-3p) and transducin β-like 1X related protein 1 (TBL1XR1). Cell counting kit-8, 5-ethynyl-2'-deoxyuridine, and flow cytometry assessed cell proliferation and apoptosis. Concentrations of inflammatory factors were examined with enzyme linked immunosorbent assay kits. Dual-luciferase reporter assay and RNA immunoprecipitation were adopted to analyze binding between miR-370-3p and circ_0035292 or TBL1XR1.
Results:
Circ_0035292 level was increased in IP patients and LPS-triggered WI-38 cells. Circ_0035292 knockdown rescued LPS-mediated WI-38 cell proliferation suppression and WI-38 cell apoptosis and inflammation promotion. Circ_0035292 interacted with miR-370-3p and miR-370-3p directly targeted TBL1XR1. Moreover, miR-370-3p overexpression alleviated LPS-induced WI-38 cell apoptosis and inflammatory injury, which was abrogated via TBL1XR1 upregulation. Circ_0035292 absence inhibited the NF-κB pathway.
Conclusion:
Knockdown of circ_0035292 rescued LPS-triggered WI-38 cell injury via miR-370-3p/TBL1XR1 axis and NF-κB pathway.
Insights
Circular RNA circ_0035292 exacerbates infantile pneumonia by promoting cell injury via the miR-370-3p/TBL1XR1 axis and NF-κB pathway. Reducing circ_0035292 levels protects against lipopolysaccharide-induced lung cell damage.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Background:
- Circular RNAs (circRNAs) are key players in human disease pathogenesis, including infantile pneumonia (IP).
- This study investigates the role of circ_0035292 in lipopolysaccharide (LPS)-induced injury in human lung cells (WI-38).
Purpose of the Study:
- To elucidate the function and mechanism of circ_0035292 in LPS-treated WI-38 cells.
- To explore the regulatory network involving circ_0035292, microRNA-370-3p (miR-370-3p), and transducin β-like 1X related protein 1 (TBL1XR1) in lung injury.
Main Methods:
- Quantitative real-time PCR and Western blot to measure molecule levels.
- Cell counting kit-8, EdU assay, and flow cytometry for proliferation and apoptosis.
- ELISA for inflammatory factors; dual-luciferase and RNA immunoprecipitation assays for molecular interactions.
Main Results:
- Circ_0035292 expression was elevated in IP patients and LPS-treated WI-38 cells.
- Knockdown of circ_0035292 reversed LPS-induced suppression of proliferation, apoptosis, and inflammation.
- Circ_0035292 sponged miR-370-3p, which targeted TBL1XR1; miR-370-3p overexpression protected cells, but this was abolished by TBL1XR1 upregulation. Circ_0035292 absence inhibited the NF-κB pathway.
Conclusions:
- Circ_0035292 knockdown mitigates LPS-induced WI-38 cell injury.
- The protective effect is mediated through the circ_0035292/miR-370-3p/TBL1XR1 axis and inhibition of the NF-κB pathway.
