Circulating differential miRNAs profiling and expression in hexavalent chromium exposed electroplating workers.

Hexavalent chromium [Cr (Ⅵ)] has extensive applications in industries, and long-term occupational exposure to Cr (Ⅵ) may lead to lung carcinoma and other cancers. While microRNA (miRNA) can take part in carcinogenesis, little is known about its expression profile in the population with Cr (Ⅵ) exposure. Thus, this study aimed to explore miRNA expression profiles in Cr (Ⅵ) exposed workers and to identify the potential biological function of differentially expressed miRNAs. A total of 45 significant differentially expressed miRNAs were identified by the miRNA array. The results of validation showed that miR-19a-3p, miR-19b-3p, and miR-142-3p were downregulated and miR-590-3p and miR-941 were upregulated in the exposure group. Multivariate analysis demonstrated that age, exposure duration and urinary chromium level were associated with one or more miRNAs expression. Target gene analysis indicated that these miRNAs might participate in the regulation of DNA damage-related signaling pathways. Taken together, Cr (Ⅵ) exposure can result in differential expression of miRNAs in occupational workers, and the expression of these miRNAs is correlated with the level and duration of Cr (Ⅵ) exposure, and the differentially expressed miRNAs may participate in DNA damage response.

Ribosomal DNA copy number is associated with P53 status and levels of heavy metals in gastrectomy specimens from gastric cancer patients.

The ribosomal DNA (rDNA) can act as a sensor and responder of cancer-associated stress. Here we investigated rDNA copy number in gastric cancers and its association with existing biomarkers and metals exposure. This study was performed on paired tumor and adjacent normal tissues obtained from 65 gastric cancer patients who underwent gastrectomy. Immunohistochemistry was used to assess HER-2, E-cadherin, EGFR, CK (pan), CK20, CK7, TopoⅡ, CAM5.2, P53, and Ki-67 expression. Inductively coupled plasma mass spectrometry (ICP-MS) was used to detect the concentrations of 17 metals in gastric tissues. rDNA copy number was detected by qPCR in genomic DNA isolated from tissue samples. Associations between the expression of existing markers, metal concentrations, and rDNA copy number were evaluated. Within patients with gastric cancer, the copy number of the 45S rDNA components (18S, 5.8S, 28S) and the 5S rDNA in tumor tissues were significantly higher than those in adjacent normal tissues, whereas mitochondrial DNA (mtDNA) copy number was significantly lower in tumor tissues than that in adjacent normal tissues. Further analysis revealed that the increase in 18S, 5.8S, and 28S rDNA copy number in tumor tissues was diminished in the context of EGFR and P53 loss. Moreover, analysis of metals revealed particularly high concentrations of As, Cd, Cr, Cu and Fe in the gastric tissues of these patients. Intriguingly, rDNA copy number variation across individuals was correlated with the concentrations of some metals. The rDNA was amplified in tumor tissues of gastric cancer patients, and its amplification may be associated with metals exposure. The expression of EGFR and P53 may influence rDNA copy number, with diminished amplification of the rDNA in cancers that were negative for these biomarkers. Our observation further our understanding of rDNA copy number in gastric cancer and its potential as a simple and useful marker in gastric cancer monitoring.

Novel DNA methylation biomarkers for hexavalent chromium exposure: an epigenome-wide analysis.

Aim: We aimed to identify differential methylation of genes that could illuminate the biological mechanisms of chromium (VI) toxicity in this exposure-control study. Materials & methods: DNA methylation was measured in blood samples collected from electroplating workers and controls using a combination of Infinium Methylation450K Chip and targeted-bisulfite sequencing. QuantiGene assay was used to detect the mRNA expression of differentially methylated genes. Inductively coupled plasma-mass spectrometry was used to quantify metals in blood and urine samples. The cytosine-phosphate-guanine sites methylation and gene expression were confirmed in a human lymphoblastoid cell line. Results & conclusion: A total of 131 differentially methylated cytosine-phosphate-guanine sites were found between exposures and controls. DNA methylation of SEMA4B may serve as a potential biomarker for chromium (VI) exposure.

Decreased 8-oxoguanine DNA glycosylase 1 (hOGG1) expression and DNA oxidation damage induced by Cr (VI).

Occupational exposure to Cr (VI) can cause DNA damage, genetic instability and elevate the risk of cancer. Here we investigated Cr (VI)-induced DNA damage and 8-oxoguanine DNA glycosylase 1 (hOGG1) gene expression in electroplating workers. The hOGG1 gene encodes a DNA repair enzyme that is crucial in DNA oxidation damage repair. Deficiency in hOGG1 DNA repair capacity contributes to the accumulation of DNA damage and genetic instability. To address the issues, we collected peripheral blood samples and urine samples from 162 electroplating workers and 84 control subjects. We measured blood chromium levels, urine chromium levels, DNA damage, and hOGG1 mRNA expression. We found significantly higher levels of blood chromium, urine chromium, and DNA damage in electroplating workers compared with controls, whereas mRNA levels of the hOGG1 gene were significantly lower in the exposed workers. Furthermore, in electroplating workers we found that blood Cr had a positive association with DNA damage as measured with the tail DNA%. Meanwhile, tail DNA% was positively associated with hOGG1 mRNA expression. Finally, the effect of potassium dichromate treatment was investigated in a human B lymphoblastoid cell line (LCL). We observed that potassium dichromate induced a concentration-dependent decrease in hOGG1 mRNA. After removing the K2Cr2O7-containing medium for 3 days and 7 days, the abundance of hOGG1 mRNA expression recovered to a similar level as the controls. Collectively, our findings suggest that decreased hOGG1 mRNA expression in occupationally exposed populations partially contribute to Cr (VI) induced DNA damage.

Elevated tissue Cr levels, increased plasma oxidative markers, and global hypomethylation of blood DNA in male Sprague-Dawley rats exposed to potassium dichromate in drinking water.

Hexavalent chromium [Cr (VI)] is prevalent in ground water in some areas, but evidence on the toxic effects of Cr (VI) via ingestion through drinking water remains insufficient. The aims of our study were to investigate the toxic effects of Cr (VI) through oral water ingestion on oxidative stress and DNA methylation. Thirty-two Sprague-Dawley rats were randomly divided into four groups, and exposed to porassium dichromate (K2 Cr2 O7 ; 0, 30, 100, and 300 mg/L) in drinking water for 4 weeks. Mean body weight gain, mean water consumption, clinical chemistry determinations, and oxidative stress levels in plasma were measured. Global DNA methylation changes and DNA methylation status at the promoter of p16 gene were also detected. After 4 weeks, mild anemic effects and increased plasma malondialdehyde (MDA) levels occurred in rats exposed to 100 mg/L or 300 mg/L of Cr (VI). Plasma glutathione peroxidase (GSH-Px) activity decreased in all exposed groups. Global DNA methylation levels were reduced in 100 mg/L and 300 mg/L exposure groups. However, DNA methylation status at the promoter of P16 gene remained unchanged in all K2 Cr2 O7- treated groups. The correlation analysis indicated that increased MDA levels were closely correlated to global DNA hypomethylation. Our results indicated that oral ingestion of Cr (VI) through drinking water caused not only oxidative stress in plasma, but also global DNA hypomethylation in blood cells from male rats, and a good correlation was found between increased MDA levels and reduced global DNA methylation. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1080-1090, 2016.

Role of DNA methylation in cell cycle arrest induced by Cr (VI) in two cell lines.

Hexavalent chromium [Cr(IV)], a well-known industrial waste product and an environmental pollutant, is recognized as a human carcinogen. But its mechanisms of carcinogenicity remain unclear, and recent studies suggest that DNA methylation may play an important role in the carcinogenesis of Cr(IV). The aim of our study was to investigate the effects of Cr(IV) on cell cycle progress, global DNA methylation, and DNA methylation of p16 gene. A human B lymphoblastoid cell line and a human lung cell line A549 were exposed to 5-15 µM potassium dichromate or 1.25-5 µg/cm² lead chromate for 2-24 hours. Cell cycle was arrested at G1 phase by both compounds in 24 hours exposure group, but global hypomethylation occurred earlier than cell cycle arrest, and the hypomethylation status maintained for more than 20 hours. The mRNA expression of p16 was significantly up-regulated by Cr(IV), especially by potassium dichromate, and the mRNA expression of cyclin-dependent kinases (CDK4 and CDK6) was significantly down-regulated. But protein expression analysis showed very little change of p16 gene. Both qualitative and quantitative results showed that DNA methylation status of p16 remained unchanged. Collectively, our data suggested that global hypomethylation was possibly responsible for Cr(IV)-induced G1 phase arrest, but DNA methylation might not be related to up-regulation of p16 gene by Cr(IV).