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Differential Expression of miR-21-5p, miR-20a-5p, TGF-β1, and TGF-β Receptor 2 in Skin, Serum, and Lung Samples
Mohadeseh Valizadeh1, Shiva Irani2, Mahmood Tavallaei3
1Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. mohadeseh_vali@yahoo.com.
Abstract:
Sulfur mustard (SM) or mustard gas is a blister chemical agent that causes pulmonary damage by triggering inflammation and oxidative injury. Alterations in microRNA (miR) transcript levels are found in pulmonary diseases and even inflammation. Therefore, we evaluated the expression levels of miR-20a-5p, miR-21-5p, and two target transcripts (transforming growth factor-beta [TGF-β1] and TGF-β receptor 2 [TGFR2]) in lung, serum, and skin samples from patients exposed to SM. Total RNA was extracted from lung, serum, and skin samples of patients with moderate (n=10) and high (n=10) SM exposure, as well as 10 healthy subjects. Following the synthesis of complementary deoxyribonucleic acid using real-time polymerase chain reaction, we determined the expression levels of miR-20a-5p, miR-21-5p, TGF-β1, and TGFR2 transcripts. Furthermore, we evaluated the sensitivity and specificity of the chosen miRs by employing receiver operating characteristic (ROC) curves and calculating the area under the ROC curve. The results showed that miR-20a-5p and miR-21-5p expressions in the groups with moderate and high SM exposure were significantly lower than the normal controls. The expression analysis demonstrated that TGFR2 was significantly less expressed in skin samples exposed to SM in both groups of patients compared with healthy controls. Furthermore, the TGF-β1 expression in the skin samples of the group with moderate SM exposure was lower than that of the normal control group. Our findings suggest that miR-20a-5p, miR-21-5p, TGF-β1, and TGFR2 expressions could be used as potential biomarkers for discriminating SM-exposed patients from healthy individuals.
Insights
Sulfur mustard (SM) exposure significantly alters microRNA (miR) and transforming growth factor-beta (TGF-β) transcript levels in patients. These specific miR and TGF-β expressions show potential as biomarkers for identifying SM-exposed individuals.
Area of Science:
- Toxicology
- Molecular Biology
- Biochemistry
Background:
- Sulfur mustard (SM) is a chemical agent causing severe pulmonary damage via inflammation and oxidative stress.
- MicroRNA (miR) transcript level alterations are linked to pulmonary diseases and inflammation.
- Investigating SM's impact on specific miRs and their targets is crucial for understanding its pathogenesis.
Purpose of the Study:
- To evaluate the expression levels of miR-20a-5p, miR-21-5p, and their target transcripts TGF-β1 and TGFR2 in patients exposed to SM.
- To determine if these molecular markers can differentiate SM-exposed individuals from healthy controls.
- To assess the diagnostic potential of these miRs using receiver operating characteristic (ROC) curve analysis.
Main Methods:
- Extraction of total RNA from lung, serum, and skin samples of moderate SM exposure (n=10), high SM exposure (n=10), and healthy (n=10) subjects.
- Complementary DNA synthesis followed by real-time polymerase chain reaction to quantify miR-20a-5p, miR-21-5p, TGF-β1, and TGFR2 transcript levels.
- Receiver operating characteristic (ROC) curve analysis to evaluate the sensitivity and specificity of the selected miRs.
Main Results:
- miR-20a-5p and miR-21-5p expression levels were significantly lower in moderate and high SM-exposed groups compared to controls.
- TGFR2 expression was significantly reduced in the skin of SM-exposed patients (both groups) versus healthy controls.
- TGF-β1 expression was lower in the skin of the moderate SM exposure group compared to controls.
Conclusions:
- Downregulation of miR-20a-5p, miR-21-5p, TGF-β1, and TGFR2 in specific tissues suggests their involvement in SM-induced pathology.
- These molecular markers hold promise as potential biomarkers for the early detection and diagnosis of SM exposure.
- Further research is warranted to validate these findings and explore their clinical utility in managing SM casualties.
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