Abstract
Objectives:To determine differential expression of microRNAs (miRNAs) in plasma of 2, 4, 6-Trinitrotoluene (TNT) exposed ordnance factory workers.Methods:A case control study was conducted at the Department of Toxicology, Armed Forces Institute of Pathology, Rawalpindi from July to December 2020. A total 30 subjects were recruited from an ordnance factory that were directly exposed to TNT and 120 non-exposed individuals from non-factory healthy population. Plasma levels of five miRNAs including miRNA-let-7a-2, miRNA-34a-1, miRNA-21-2, miRNA-106b-1, miRNA-122a-1 were measured by quantitative real-time polymerase chain reaction (RT-PCR).Results:Micro RNAs showed a wide range of Ct (cycle threshold) values ranging from 23.48 to 41.94. Among the five miRNAs let-7a-2 and miRNA-122a-1 displayed relatively high expression with Ct values ranging from 26.58 ± 2.25 to 27.18 ± 0.80 respectively. Relative fold change expression for all five miRNAs of exposed individuals were found high (p <0.0001) vs non-exposed. Dividing fold change expression of exposed individuals into two groups as ≤ 10 and > 10, the individuals having ≤ 10-fold change expression were19 (63.3%) in miRNA-let-7a-2, 30 (100%) in miRNA-34a-1 and 23 (76.7%) in miRNA-122a-1 while in miRNA-21-2 and miR-106b-1, 23 (76.7%) and 18(60%) individuals had > 10-fold change expression respectively. Among the five miRNAs in exposed individuals, miRNA-let-7a-2, miR-21-2, miR-106b-1 and miR-122a-1 were found highly expressed with fold change expression > 10 (p <0.0001). No significant association was found between miRNAs expression levels with age and working duration.Conclusion:The study shows upregulation of all five miRNAs in TNT exposed subjects with no significant association of expression levels with age and working duration.
Highlights
Two, 4, 6 Trinitrotoluene (TNT) is a yellow, odorless solid that has been commonly used as an explosive for military and industrial purposes since the beginning of the twentieth century[1] and considered a serious environmental pollutant posing a grave risk to occupationally exposed population
A total of 30 cases working in Ordnance factory with ages ranging from 25–56 years and working duration ranging from 4 – 34 were included in the study
Five miRNAs were evaluated by assay specific real-time PCR assay after normalization with equivalence unit mass of total RNA extracted from equal volumes of plasma for each sample
Summary
4, 6 Trinitrotoluene (TNT) is a yellow, odorless solid that has been commonly used as an explosive for military and industrial purposes since the beginning of the twentieth century[1] and considered a serious environmental pollutant posing a grave risk to occupationally exposed population. Considering association of TNT and its metabolites with different types of cancers, there is a need to identify markers for early detection of cancer in occupationally exposed persons to TNT. MicroRNAs (miRNAs) have come up as a promising diagnostic modality that discriminates different malignancies from their normal counterparts in early stages of cancer.[4] They are short (19 to 25 nucleotides base-pairs), single stranded non-coding RNAs that regulate gene expression and cellular processes – differentiation, apoptosis and proliferation.[5] Defective expression of some miRNAs that are associated with cancers, like hepatocellular carcinoma, leukemia and bladder cancer include miRNA-122, miRNA 21, miRNA let-7, miRNA-34a and miRNA -106.6-10
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