miRNA-34b is directly involved in the aging of macrophages
Wei Liang1, Sheng Gao1, Liu Liang1
1National Center for International Research of Biological Targeting Diagnosis and Therapy, Guangxi Key Laboratory of Biological Targeting Diagnosis and Therapy Research, Collaborative Innovation Center for Targeting Tumor Diagnosis and Therapy, Guangxi Medical University, Nanning, Guangxi, 530021, China.
Background:
MicroRNAs (miRNAs) are a class of short noncoding RNA that play important regulatory roles in living organisms. These RNA molecules are implicated in the development and progression of malignant diseases such as cancer and are closely associated with cell aging. Findings demonstrating that microRNA is associated with aging in macrophages have nevertheless rarely been reported.
Aims:
This study's objective was to investigate if miRNA-34 is linked to aging process of macrophages.
Methods:
We built a cell aging model in mouse RAW264.7 macrophages using D-galactose and determined the expression levels of miRNA-34a, miRNA-34b, and miRNA-34c in aging and normal macrophages by fluorescence quantitative polymerase chain reaction (q-PCR). We predicted a target gene of miRNA-34 using biological information techniques and constructed the recombinant plasmid pGL3-E2f3 for the putative target gene E2f3.
Results:
The expression level of miRNA-34b was 5.23 times higher in aging macrophages than in normal macrophages. The luciferase activity decreased by nearly 50 % in cells transfected with miRNA-34b mimics, while no significant decrease in luciferase activity was noted in cells transfected with the miRNA-34b inhibitor or unrelated sequences.
Discussion:
Our findings provide the groundwork for further research into the molecular mechanisms whereby miRNA-34b regulates the aging of macrophages.
Conclusions:
miRNA-34b is associated with the aging of RAW264.7 macrophages, and E2f3 is a target gene of miRNA-34b.
Insights
MicroRNA-34b is significantly elevated in aging macrophages, suggesting a role in the cellular aging process. This study identifies E2f3 as a direct target of microRNA-34b in macrophages.
Area of Science:
- Molecular Biology
- Cellular Aging Research
- Biochemistry
Background:
- MicroRNAs (miRNAs) are short noncoding RNAs involved in regulating gene expression.
- miRNAs are implicated in aging and diseases like cancer.
- The association between miRNAs and macrophage aging is not well-established.
Purpose of the Study:
- To investigate the link between miRNA-34 family members and macrophage aging.
- To determine if miRNA-34b expression changes during macrophage aging.
Main Methods:
- A cell aging model was established in mouse RAW264.7 macrophages using D-galactose.
- Quantitative PCR (q-PCR) was used to measure miRNA-34a, miRNA-34b, and miRNA-34c expression.
- Bioinformatics and luciferase assays were employed to identify and validate miRNA-34b targets, including E2f3.
Main Results:
- miRNA-34b expression was 5.23-fold higher in aging macrophages compared to normal macrophages.
- Transfection with miRNA-34b mimics reduced luciferase activity by approximately 50%, confirming E2f3 as a target.
- No significant changes in luciferase activity were observed with miRNA-34b inhibitors or control sequences.
Conclusions:
- miRNA-34b is significantly associated with the aging process in RAW264.7 macrophages.
- E2f3 is identified as a direct target gene regulated by miRNA-34b.
- These findings lay the foundation for understanding miRNA-34b's role in macrophage aging mechanisms.
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