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Effects of quercetin on CDK4 mRNA and protein expression in A549 cells infected by H1N1
Qiaofeng Wan1, Hao Wang1, Yuan Lin1
1Department of Pathogen Biology and Immunology, Basic Medical Science College, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China.
Abstract:
This study was conducted to investigate the effects of quercetin on the expression of cyclin-dependent kinase (CDK4) mRNA and protein in A549 lung epithelial tumor cells infected by H1N1. First, the Thiazolyl Blue Tetrazolium Bromide (MTT) method was used to determine H1N1 virulence, quercetin cytotoxicity and inhibition of the cytopathic effect of H1N1 on A549 cells by quercetin. Subsequently, 100 TCID50 H1N1 was used to infect A549 cells for 2 h prior to culture in maintenance media containing 10 mg/l quercetin. After 4, 12, 24 and 48 h of culture, the cells were collected and total RNA and protein were extracted. Fluorescent quantitative polymerase chain reaction and western blot analysis were then performed to assess the expression of CDK4 mRNA and protein. The experiment demonstrated that the TCID50 of H1N1 in A549 cells was 10-4.75, the maximum non-toxic concentration of quercetin in A549 cells was 30-60 mg/l and the minimum effective concentration of quercetin for the inhibition of the H1N1 cytopathic effect on A549 cells was 10 mg/l. The results indicated that quercetin may significantly inhibit CDK4 mRNA and protein overexpression caused by H1N1 within 4-48 h. In conclusion, quercetin may protect against H1N1 infection by effectively reducing the mRNA and protein expression of CDK4 caused by H1N1 infection.
Insights
Quercetin effectively reduces cyclin-dependent kinase 4 (CDK4) mRNA and protein levels in A549 lung cells during H1N1 influenza virus infection. This natural compound shows potential in protecting against H1N1 by mitigating viral-induced CDK4 overexpression.
Area of Science:
- Molecular Biology
- Virology
- Pharmacology
Background:
- Influenza A (H1N1) virus infection poses a significant global health threat.
- Lung epithelial tumor cells (A549) are a relevant model for studying viral pathogenesis.
- Cyclin-dependent kinase 4 (CDK4) is implicated in cellular proliferation and viral replication.
Purpose of the Study:
- To investigate the impact of quercetin on CDK4 mRNA and protein expression in H1N1-infected A549 cells.
- To determine the antiviral efficacy of quercetin against H1N1 cytopathic effects in A549 cells.
Main Methods:
- Cytotoxicity and antiviral activity of quercetin assessed using the Thiazolyl Blue Tetrazolium Bromide (MTT) assay.
- H1N1 infection of A549 cells followed by treatment with quercetin.
- Quantitative real-time PCR (qRT-PCR) and Western blot analysis to measure CDK4 mRNA and protein levels.
Main Results:
- Established H1N1 virulence (100 TCID50), quercetin cytotoxicity (30-60 mg/l), and minimum effective concentration (10 mg/l).
- Quercetin significantly inhibited H1N1-induced overexpression of CDK4 mRNA and protein within 4 to 48 hours post-infection.
- Observed reduction in viral cytopathic effects in quercetin-treated cells.
Conclusions:
- Quercetin demonstrates a protective effect against H1N1 infection in A549 lung epithelial cells.
- The mechanism involves the significant reduction of H1N1-induced CDK4 mRNA and protein expression.
- Quercetin represents a potential therapeutic agent for managing H1N1 influenza.
