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Published on: March 14, 2017
[Identification of microRNAs targeting vitamin D receptor and their effect on parathyroid hormone secretion in
1Department of Breast and Thyroid Surgery, Third Xiangya Hospital of Central South University, Changsha 410013, China.
Objective:
To identify the miRNAs targeting vitamin D receptor (VDR) gene and their effect on parathyroid hormone (PTH) secretion in secondary hyperparathyroidism.
Methods:
Primary parathyroid cells with secondary hyperparathyroidism were isolated by collagenase digestion and cultured. The miRNAs targeting VDR were screened by bioinformatics methods and full transcriptome sequencing, and dual-luciferase reporter assay was used to verify the targeting relationship between VDR and the screened miRNA. The effects of overexpression or inhibition of the candidate miRNA on VDR mRNA and protein expressions and PTH secretion were evaluated using qRT-PCR and Western blotting. The expression levels of the candidate miRNAs and VDR mRNA in clinical specimens of parathyroid tissues were verified by qRT-PCR, and the expression of VDR protein was detected by immunohistochemistry.
Results:
We successfully isolated primary parathyroid cells. Dual-luciferase reporter assay verified the targeting relationship of hsa-miR-149-5p, hsa-miR-221-5p, hsa-miR-222-3p, hsa-miR-29a-5p, hsa-miR-301a-5p, hsa-miR-873-5p, hsa-miR-93-3p with VDR, and among them, the overexpression of hsa-miR-149-5p and hsa-miR-301a-5p significantly increased PTH secretion in the parathyroid cells. In patients with secondary hyperparathyroidism, hsa-miR-149-5p was highly expressed in the parathyroid tissues (P=0.046), where the expressions of VDR mRNA (P=0.0267) and protein were both decreased.
Conclusion:
The two miRNAs, hsa-miR-149-5p and hsa-miR-301a-5p, may promote the secretion of PTH in patients with secondary hyperparathyroidism by down-regulating the expression of VDR gene.
Insights
Two miRNAs, hsa-miR-149-5p and hsa-miR-301a-5p, were found to increase parathyroid hormone (PTH) secretion by down-regulating the vitamin D receptor (VDR) gene in secondary hyperparathyroidism.
Area of Science:
- Endocrinology
- Molecular Biology
- Genetics
Background:
- Secondary hyperparathyroidism is characterized by elevated parathyroid hormone (PTH) levels.
- The vitamin D receptor (VDR) plays a crucial role in regulating PTH secretion.
- MicroRNAs (miRNAs) are implicated in various endocrine disorders, including secondary hyperparathyroidism.
Purpose of the Study:
- To identify specific miRNAs that target the VDR gene.
- To investigate the impact of these miRNAs on PTH secretion in secondary hyperparathyroidism.
- To elucidate the molecular mechanisms underlying VDR regulation by miRNAs in this condition.
Main Methods:
- Isolation and culture of primary parathyroid cells from patients with secondary hyperparathyroidism.
- Bioinformatic screening and full transcriptome sequencing to identify VDR-targeting miRNAs.
- Dual-luciferase reporter assays to confirm miRNA-VDR interactions.
- Quantitative real-time PCR (qRT-PCR) and Western blotting to assess VDR expression and PTH secretion.
- Analysis of miRNA and VDR expression in clinical parathyroid tissue samples.
Main Results:
- Dual-luciferase assays confirmed that hsa-miR-149-5p, hsa-miR-301a-5p, and other miRNAs target the VDR gene.
- Overexpression of hsa-miR-149-5p and hsa-miR-301a-5p significantly increased PTH secretion.
- In patients with secondary hyperparathyroidism, hsa-miR-149-5p was upregulated, while VDR mRNA and protein expression were decreased in parathyroid tissues.
Conclusions:
- hsa-miR-149-5p and hsa-miR-301a-5p are identified as key miRNAs targeting the VDR gene.
- These miRNAs may promote PTH secretion in secondary hyperparathyroidism by down-regulating VDR expression.
- Targeting these miRNAs could offer a novel therapeutic strategy for secondary hyperparathyroidism.
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