Association of LncRNA-PAX8-AS1 and LAIR-2 polymorphisms along with their expression with clinical and subclinical hypothyroidism

Abstract

The genetic and epigenetic architecture of clinical and subclinical hypothyroidism remains unclear. We investigated the impact of long noncoding RNA (LncRNA)-PAX8-AS1 and LAIR-2 genetic variants on the susceptibility to clinical and subclinical hypothyroidism, their influence on LncRNA-PAX8-AS1 and LAIR-2 expression and their potential as hypothyroid biomarkers. Hundred clinical hypothyroid patients, 110 subclinical hypothyroid patients, and 95 healthy controls were enrolled. Gene expression analysis and genotyping were performed by qPCR. LAIR-2 protein, a proinflammatory mediator, was tested by ELISA. Serum LncRNA-PAX8-AS1 was downregulated, whereas LAIR-2 mRNA and protein levels were upregulated in clinical and subclinical hypothyroid patients compared to healthy controls. LncRNA-PAX8-AS1 rs4848320 and rs1110839 were associated with increased risk of clinical hypothyroidism. Interestingly, both SNPs were associated with differential expression of serum LncRNA-PAX8-AS1 among clinical hypothyroid patients. LAIR-2 rs2287828 was associated with elevated risk of both clinical and subclinical hypothyroidism. Harboring the rs2287828 T allele augmented the LAIR-2 mRNA expression among clinical hypothyroid patients, while elevated both LAIR-2 mRNA and protein levels in subclinical hypothyroid patients. The rs4848320-rs1110839-rs2287828 TTT, CTT, and CGT haplotypes were associated with increased hypothyroid risk. Surprisingly, serum LncRNA-PAX8-AS1 and LAIR-2 mRNA expression demonstrated superior diagnostic accuracy for clinical hypothyroidism and turned out as independent predictors in the multivariate analysis. Conclusively, LncRNA-PAX8-AS1 and LAIR-2 genetic variants are novel genetic biomarkers of hypothyroidism that could alter the LncRNA-PAX8-AS1 and LAIR-2 expression. LncRNA-PAX8-AS1 and LAIR-2 expression profiles have the potential as effective diagnostic and prognostic indicators of hypothyroidism.

Association of lncRNA MEG3 rs941576 polymorphism, expression profile, and its related targets with the risk of obesity-related colorectal cancer: potential clinical insights

“…The cDNA samples were appropriately diluted and amplified using the miScript SYBR Green PCR kit (Qiagen) and the supplied miScript Universal Primer (reverse primer), along with ready-made specific miScript Primer (forward primer) Assays for hsa-miR-27a-3p, hsa-miR-181a-5p, and the housekeeping miScript PCR control miRNA SNORD68 (Qiagen). SNORD68 was validated as an internal control for normalization of miRNAs in several studies which supported its use as a reference for miRNA relative quantification based on its stable and equivalent expression between the sera of diseased patients and controls 20 , 44 , 46 . Briefly, real-time PCR was performed in 20 μL reaction mixtures prepared as previously described 20 , 46 using the Qiagen Rotor-Gene Q system.…”

“…SNP genotyping was attempted using the real-time PCR TaqMan allelic discrimination assay. Amplification of DNA was conducted as previously described 20 , 44 using the Qiagen Rotor-Gene Q Real-time PCR System by utilizing a TaqMan Master Mix and pre-designed primer/probe sets for rs941576 (A/G) [Catalog number: 4351379] (Thermo Scientific, USA). The PCR thermal cycler conditions (95 °C for ten minutes, then 40 cycles of 92 °C for fifteen s and 60 °C for ninety seconds) were implemented.…”

“…Primer sequences are mentioned in Table 1 . In short, qPCR was carried out on the Qiagen Rotor-Gene Q System using twenty μL reaction mixtures prepared using the PCR Maxima SYBR Green kit (Thermo Scientific, USA) along with forward and reverse primers as previously described 20 , 44 . The thermal cycler runs were ten minutes at 95 °C, followed by 45 cycles of fifteen seconds at 95 °C and sixty seconds at 60 °C.…”

Serum lncRNAs TUG1, H19, and NEAT1 and their target miR-29b/SLC3A1 axis as possible biomarkers of preeclampsia: Potential clinical insights

“…Using GAPDH as an internal control gene, the expression profiles of TUG1, H19, NEAT1, and SLC3A1 were determined by qPCR. The GAPDH gene has been validated previously for use as an excellent internal control for normalizing lncRNAs [ 43 , 44 ]. The qPCR assay was conducted using the Maxima SYBR Green PCR kit (ThermoFischer, USA) and customized primers (Metabion, Germany) following the manufacturer's recommendations.…”