[微小RNA-301a-5p可能通过调节钙敏感受体来调控继发性甲状旁腺功能亢进中的甲状旁腺激素分泌]

[MiR-301a-5p modulates parathyroid hormone secretion in secondary hyperparathyroidism possibly by regulating calcium-sensing receptor].

作者信息

Liu L, Qian L, Li P, Li J, Huang S, Yi W, Liu S, Wu W

机构信息

Department of Breast and Thyroid Surgery, Third Xiangya Hospital, Central South University, Changsha 410013, China.

Department of Breast and Thyroid Surgery, Second Xiangya Hospital, Central South University, Changsha 410013, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2023 Aug 20;43(8):1363-1370. doi: 10.12122/j.issn.1673-4254.2023.08.13.

DOI:10.12122/j.issn.1673-4254.2023.08.13

PMID:37712273

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10505570/

Abstract

OBJECTIVE

To explore the miRNAs that down- regulate calcium-sensing receptor (CaSR) in secondary hyperparathyroidism (SHPT) and their effects on parathyroid hormone (PTH) secretion.

METHODS

Whole transcriptome sequencing was performed for 6 normal parathyroid tissue samples and 11 SHPT parathyroid tissue samples. Based on bioinformatic prediction, we screened out 7 candidate miRNAs that regulate CaSR, among which the most likely miRNA for CaSR regulation was identified by double luciferase test. We detected the differential expression of miR-301a-5p and CaSR mRNA in SHPT and normal parathyroid tissue using qRT-PCR, and analyzed the correlation between their expressions and serum PTH levels of the patients. Western blotting was used to detect the expression of CaSR protein in primary SHPT parathyroid cells transfected with miR-301a-5p mimics or inhibitors, and the level of PTH in the supernatant of the cell culture was determined.

RESULTS

Among the preliminarily selected 7 miRNAs that potentially regulate CaSR (miR-15a-5p, miR-15b-5p, miR- 16- 5p, miR- 221- 3p, miR- 222- 3p, miR- 301a- 5p and miR- 503- 5p), miR- 301a-5p was significantly upregulated in SHPT compared with normal parathyroid tissue ( < 0.05), and its expression appeared to be positively correlated with PTH level, but this correlation was not statistically significant ( > 0.05); The expression of CaSR mRNA was significantly downregulated in SHPT ( < 0.05), and its expression tended to inversely correlate with the patient's PTH level, but the correlation was not statistically significant ( > 0.05). In primary culture of SHPT parathyroid cells, miR-301a-5p overexpression caused a significant decrease of CaSR protein expression ( < 0.05), and conversely, inhibition of miR-301a-5p expression increased the expression of CaSR protein ( < 0.05). Although miR-301a-5p overexpression did not significantly affect PTH secretion of the cells ( > 0.05), inhibition of iR-301a-5p expression strongly increased the secretion of PTH ( < 0.05).

CONCLUSION

MiR-301a-5p affects PTH secretion in SHPT possibly by regulating the expression of CaSR.

摘要

目的

探讨继发性甲状旁腺功能亢进症（SHPT）中下调钙敏感受体（CaSR）的微小RNA（miRNA）及其对甲状旁腺激素（PTH）分泌的影响。

方法

对6例正常甲状旁腺组织样本和11例SHPT甲状旁腺组织样本进行全转录组测序。基于生物信息学预测，筛选出7个调控CaSR的候选miRNA，通过双荧光素酶试验确定其中最有可能调控CaSR的miRNA。采用qRT-PCR检测SHPT和正常甲状旁腺组织中miR-301a-5p和CaSR mRNA的差异表达，并分析它们的表达与患者血清PTH水平的相关性。用蛋白质免疫印迹法检测转染miR-301a-5p模拟物或抑制剂的原发性SHPT甲状旁腺细胞中CaSR蛋白的表达，并测定细胞培养上清液中PTH的水平。

结果

在初步筛选出的7个可能调控CaSR的miRNA（miR-15a-5p、miR-15b-5p、miR-16-5p、miR-221-3p、miR-222-3p、miR-301a-5p和miR-503-5p）中，与正常甲状旁腺组织相比，miR-301a-5p在SHPT中显著上调（<0.05），其表达似乎与PTH水平呈正相关，但这种相关性无统计学意义（>0.05）；CaSR mRNA的表达在SHPT中显著下调（<0.05），其表达倾向于与患者的PTH水平呈负相关，但相关性无统计学意义（>0.05）。在SHPT甲状旁腺细胞原代培养中，miR-301a-5p过表达导致CaSR蛋白表达显著降低（<0.05），相反，抑制miR-301a-5p表达可增加CaSR蛋白的表达（<0.05）。虽然miR-301a-5p过表达对细胞的PTH分泌无显著影响（>0.05），但抑制miR-301a-5p表达可显著增加PTH的分泌（<0.05）。