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miR-433 Inhibits Neuronal Growth and Promotes Autophagy in Mouse Hippocampal HT-22 Cell Line
Chunli Xu1, Qingke Bai2, Chen Wang3
1Department of Neurology, The Seventh People's Hospital of Integrated Traditional Chinese and Western Medicine, Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Abstract:
Background: MicroRNAs (miRNAs) have an increasing functional role in some neurodegenerative diseases. Autophagy, the degradation of bulk protein in the cytoplasm, is the quality control function of protein and has a protective role in the survival of neural cells. miR-433 may play a regulatory role in neurodegenerative diseases. Many aspects underlying the mechanism of miR-433 in neural development and neurodegeneration are not clear. Methods: In this study, we established stable cell lines expressing miR-433 by infecting mouse hippocampal neural cell line (HT-22) cells with rLV-miR-433 and the control rLV-miR. Pre-miR-433 expression was analyzed using polymerase chain reaction (PCR). Mature miR-433 expression was measured using quantitative PCR (qPCR). The effect of miR-433 overexpression on cell proliferation was determined using a CCK-8 assay and flow cytometry. RNA interference was used to analyze the function of Cdk12 in mediating the effect of miR-433 on cell proliferation. The effect of miR-433 overexpression on cell apoptosis was determined by flow cytometry. Autophagy-related genes Atg4a, LC3B, and Beclin-1 were determined using qPCR, Western blot, or immunofluorescence. In addition, RNA interference was used to analyze the effect of Atg4a on the induction of autophagy. TargetScan 7.2 was used to predict the target genes of miR-433, and Smad9 was determined using qPCR. Results: Our results indicated that miR-433 increased the expression of Atg4a and induced autophagy by increasing the expression of LC3B-Ⅱ and Beclin-1 in an Atg4a-dependent manner. In addition, miR-433 upregulated the expression of Cdk12 and inhibited cell proliferation in a Cdk12-dependent manner and promoted apoptosis in HT-22 cells under the treatment of 10-hydroxycamptothecin. Conclusion: The results of our study suggest that miR-433 may regulate neuronal growth by promoting autophagy and attenuating cell proliferation. This might be a potential therapeutic intervention in neurodegenerative diseases.
Insights
MicroRNA-433 (miR-433) promotes neural cell survival by enhancing autophagy and inhibiting cell proliferation. This microRNA may offer a new therapeutic strategy for neurodegenerative diseases.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) play crucial roles in neurodegenerative diseases.
- Autophagy is vital for neural cell survival and protein quality control.
- The specific role of miR-433 in neurodegeneration remains unclear.
Purpose of the Study:
- To investigate the functional mechanism of miR-433 in neural cells.
- To determine the effect of miR-433 on autophagy, cell proliferation, and apoptosis.
- To explore the potential of miR-433 as a therapeutic target for neurodegenerative diseases.
Main Methods:
- Established stable miR-433 overexpressing cell lines (HT-22).
- Assessed cell proliferation, apoptosis, and autophagy markers (Atg4a, LC3B, Beclin-1).
- Utilized quantitative PCR, Western blot, immunofluorescence, and RNA interference.
Main Results:
- miR-433 overexpression induced autophagy via Atg4a, LC3B-II, and Beclin-1.
- miR-433 upregulated Cdk12, inhibiting cell proliferation in a Cdk12-dependent manner.
- miR-433 promoted apoptosis in HT-22 cells under 10-hydroxycamptothecin treatment.
Conclusions:
- miR-433 regulates neuronal growth by promoting autophagy and attenuating proliferation.
- miR-433 demonstrates potential as a therapeutic intervention for neurodegenerative diseases.
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