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Published on: June 15, 2018
LncRNA Pvt1 aggravates cardiomyocyte apoptosis via the microRNA-216/Ccnd3 axis
Yu Hu1,2, Minghao Luo1, Yuzhou Xue3
1Department of Cardiology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Objective:
Our study aims to evaluate the role of long non-coding RNA variant translocation gene Pvt1 in cardiomyocyte apoptosis, as well as the potential targets and mechanisms involved in Pvt1-miRNA-mRNA axis.
Methods:
1.Pvt1 knockdown in cells by transfection with small interfering RNA (si-Pvt1), HL-1 cells were randomly divided into control group, hypoxia group, hypoxia + negative control group and hypoxia + si-Pvt1 group. Apoptosis-related genes expression was detected by Western blot assay, RT-qPCR and Flow cytometry assay. 2.Pvt1 knockdown model (sh-Pvt1) was established by injecting adeno-associated virus (AAV) vector shRNA-Pvt1 into the caudal vein 7 days before myocardial infarction, and echocardiography was used to measure cardiac function 7 days after myocardial infarction induced by ligation of the left anterior descending branch. HE staining was used to evaluate the pathological injury of mouse heart tissue, and the apoptotic protein expression was detected by Western blot. 3.lncRNA-related microRNAs were predicted by bioinformatics tools and further verified by dual luciferase experiment. Western blot analysis was used to identify the expression of apoptotic genes following the simultaneous transfection of si-Pvt1 and miR-216 mimics. Genes differentially expressed in hypoxia + si-NC and hypoxia + si-Pvt1 groups were identified by RNA sequencing. These genes were then compared with the target genes of miR-216 predicted by bioinformatics tools. The gene of interest Ccnd3 was excluded from the analysis. Western blot analysis was used to assess the expression of Apoptosis-related proteins in HL-1 cells co-transfected with miR-216 mimics and overexpressed Ccnd3.
Results:
1. Pvt1 was highly expressed in HL-1 induced by hypoxia, and Pvt1 knockdown can reduce cell apoptosis in hypoxia cells. 2. MI causes myocardial injury in mice, and inhibition of Pvt 11 can improve the cardiac function of mice with myocardial infarction, prevent some inflammatory cell infiltration, and reduce myocardial cell apoptosis. 3. Pvt1 acts as a sponge for miR-216 and promotes the expression of Ccnd3.
Conclusion:
Pvt1 may promote myocardial infarct-induced apoptosis through the miR-216/Ccnd3 axis.
Insights
Long non-coding RNA Pvt1 promotes cardiomyocyte apoptosis via the miR-216/Ccnd3 pathway. Inhibiting Pvt1 reduces myocardial infarction injury and improves cardiac function by decreasing cell apoptosis.
Area of Science:
- Molecular Biology
- Cardiovascular Research
- Gene Regulation
Background:
- Long non-coding RNAs (lncRNAs) play critical roles in cellular processes, including apoptosis.
- Pvt1 (gene variant translocation gene Pvt1) has emerged as a key regulator in various biological functions.
- Understanding the role of Pvt1 in cardiomyocyte apoptosis is crucial for developing therapeutic strategies for myocardial infarction.
Purpose of the Study:
- To investigate the role of lncRNA Pvt1 in cardiomyocyte apoptosis.
- To elucidate the underlying molecular mechanisms, focusing on the Pvt1-miRNA-mRNA axis.
- To evaluate the therapeutic potential of targeting Pvt1 in myocardial infarction.
Main Methods:
- Pvt1 knockdown was achieved using small interfering RNA (si-Pvt1) in HL-1 cells and adeno-associated virus (AAV) vectors in a mouse model of myocardial infarction (MI).
- Cardiac function was assessed using echocardiography, and myocardial injury was evaluated by HE staining.
- Gene and protein expression of apoptosis-related factors were analyzed by Western blot, RT-qPCR, and flow cytometry.
- The interaction between Pvt1, miR-216, and Ccnd3 was confirmed using bioinformatics, dual luciferase assays, and RNA sequencing.
Main Results:
- Pvt1 expression was upregulated in hypoxia-induced HL-1 cells, and its knockdown reduced hypoxia-induced apoptosis.
- Inhibition of Pvt1 improved cardiac function, reduced inflammatory cell infiltration, and attenuated cardiomyocyte apoptosis in mice with MI.
- Pvt1 functions as a molecular sponge for miR-216, thereby promoting the expression of Ccnd3.
Conclusions:
- Pvt1 promotes myocardial infarction-induced apoptosis through the miR-216/Ccnd3 axis.
- Targeting Pvt1 offers a potential therapeutic strategy for mitigating myocardial infarction-related damage.
- The Pvt1-miRNA-mRNA axis represents a novel regulatory pathway in cardiomyocyte apoptosis.
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