Identification of Differentially Expressed Genes in Lung Tissues of Nickel-Exposed Rats Using Suppression Subtractive Hybridization

Abstract

Occupational exposure to nickel compound, such as nickel refining, electroplating, and in conjunction with other metals, is harmful to the health, causing respiratory distress, and lung and nasal cancer. In this work, the different gene expression patterns of lung tissues from nickel-exposed rats and controls were investigated. The suppression subtractive hybridization (SSH) method was used to generate two subtracted cDNA libraries with gene transcripts differentially expressed after nickel inducing. Dot-blot hybridizations were used to confirm differential ratios of expression of obtained SSH clones. Out of 768 unique SSH clones, which were chosen randomly from the two subtraction libraries (384 of each), 319 could be verified as differentially expressed. According to blast screening and functional annotation, 28% genes in nickel-induced cDNA library were related to cell differentiation, whereas 21% in driver library were related to oxygen transport. Two novel expressed sequence tags (ESTs; NCBI Accession No. FC809414 and No. FC809411) in nickel-induced cDNA library were obtained. The genes detected in the present study are probably important genes associated with nickel-induced lung cancer.

Time- and concentration-dependent genomic responses of the rat airway to inhaled nickel subsulfide

“…It is difficult to compare the gene expression responses in the present study (from lung target tissue of rats after repeated inhalation exposure) with responses from submerged transformed human lung cells at brief, orders-of-magnitude higher in vitro exposures, or with chronic responses from in vivo rat studies where a non-physiological route of exposure was employed in a different strain. However, the transition from down-regulation of proliferation in the shorter-term in vitro study (Andrew et al, 2003) to upregulation in the longer-term in vivo study (Zhang et al, 2011) is consistent with the pattern of gene changes between 1 and 4 weeks of exposure in our study. The present results support a nonlinear mode of action for the carcinogenicity of Ni 3 S 2 , and the utilization of a nonlinear approach in risk assessment for this compound rather than the default linear doseresponse option for carcinogens.…”

“…The highdose in vitro exposure resulted in down-regulation of a number of genes associated with proliferation (c-myc, Cyclin K, ERF1, hEGR1, MAPKAPK-2, TAFAP-2). Zhang et al (2011) evaluated the gene expression changes in the lungs of Wistar rats given a single intramuscular injection of a suspension of 10 mg Ni 3 S 2 . After 7 months, the animals were sacrificed and the lungs were evaluated.…”

Analysis of differentially expressed genes in two immunologically distinct strains of Eimeria maxima using suppression subtractive hybridization and dot-blot hybridization

“…Briefly, the cDNA was digested with Rsa I to remove the adaptor sequence, as reported previously [24,25], and then purified. A 1-μg aliquot of cDNA was labeled with DIG-11-2′-deoxyuridine 5″-triphosphate (Roche Applied Science) in a randomly primed DNA-labeling reaction as described by the manufacturer.…”