Abstract:Objective To study the role of latent transforming growth factor-β binding protein 2(LTBP2) gene in oxidative damage of the human bronchial epithelial cells (16HBE) induced by cadmium chloride (CdCl2). Methods 16HBE cells were treated with different doses of CdCl2 (10 μmol/L, 20 μmol/L, 30 μmol/L and 40 μmol/L) for 24 hours, and with 30 μmol/L CdCl2 for 12, 24 and 48 hours, respectively. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) assay was used to detect the changes in the expression of LTBP2. Specific shRNA sequences were designed according to the target gene. Then it was annealed and assembled on lentivirus vector pCDH. It was successfully constructed for the stable LTBP2 under-expression in 16HBE cell lines. The contents of malondialdehyde (MDA) and the activity of total superoxide dismutase (SOD) were measured by hydroxylamine method and thiobarbituric acid colorimetry, respectively. Results Compared with the control group (0 μmol/L CdCl2), it was significantly different for the relative expression of LTBP2 mRNA in 30 μmol/L and 40 μmol/L groups (both P<0.05), and the expression of LTBP2 mRNA increased gradually with the increment of CdCl2 dosage (P<0.01). Compared with the control group (0 μmol/L CdCl2), it was significantly different for the relative expression of LTBP2 mRNA in 16HBE cells which were treated with 30 μmol/L CdCl2 for 24 and 48 hours, respectively (both P<0.01). Compared with 16HBE, SOD content in 16HBE-shLTBP2 increased significantly (P<0.05), but MDA content in 16HBE-shLTBP2 decreased significantly (P<0.05) when treated with different concentration of CdCl2 for different time points. Conclusions The under-expression of LTBP2 decreased cadmium-induced oxidative damage of 16HBE cells. It is suggested that LTBP2 gene may play a promoting role in oxidative damage progress induced by CdCl2.
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