| Li Qiang,Bao Yizhong,Zhang Xuxia,Gao Yun,Ding Defang,Ren Xiangyi,Chen Honghong.Role of GSK-3β/β-catenin signaling pathway in human renal proximal tubular epithelial cell injury induced by depleted uranium[J].Chinese Journal of Radiological Medicine and Protection,2017,37(3):171-176 |
| Role of GSK-3β/β-catenin signaling pathway in human renal proximal tubular epithelial cell injury induced by depleted uranium |
| Received:November 07, 2016 |
| DOI：10.3760/cma.j.issn.0254-5098.2017.03.002 |
| KeyWords:Depleted uranium GSK-3β β-catenin KIM-1 HK-2 cell |
| FundProject:国家科技重大专项子课题（2013ZX09J13101-05B） |
| Author Name | Affiliation | E-mail | | Li Qiang | Department of Radiation Biology, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China | | | Bao Yizhong | Department of Radiation Biology, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China | | | Zhang Xuxia | Department of Radiation Biology, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China | | | Gao Yun | Department of Radiation Biology, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China | | | Ding Defang | Department of Radiation Biology, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China | | | Ren Xiangyi | Department of Radiation Biology, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China | | | Chen Honghong | Department of Radiation Biology, Institute of Radiation Medicine, Fudan University, Shanghai 200032, China | hhchen@shmu.edu.cn |
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| Abstract:: |
| Objective To investigate the effects of glycogen synthase kinase-3β (GSK-3β) and β-catenin signaling on the human renal proximal tubular epithelial HK-2 cell injury induced by depleted uranium(DU), and provide a new enlightenment for the development of DU antidotes. Methods HK-2 cells were exposed to different concentrations of DU for 3-24 h, then the protein expressions of kidney injury molecule 1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL) and nuclear β-catenin were detected by immunofluorescence staining. The protein expressions of p-GSK-3β(S9), GSK-3β and c-myc were detected by Western blot assay. HK-2 cells were transiently transfected by GSK-3β (KD) plasmid or treated by TDZD-8 to inhibit the activity of GSK-3β specifically. Other HK-2 cells were transiently transfected by β-catenin plasmid to overexpress the β-catenin protein. Results The percentages of KIM-1 and NGAL-positive cells increased with DU exposure time and concentrations from 300 and 600 μmol/L, and they were significantly higher than those of the blank control at 6-24 h of DU exposure (KIM-1-positive cells:t=11.06, 18.97, 30.49, P<0.05; t=6.79, 16.02, 85.45, P<0.05; NGAL-positive cells:t=11.78, 11.37, 34.29, P<0.05; t=7.34, 21.63, 36.84, P<0.05). In contrast, the ratio of p-GSK-3β(S9) to GSK-3β and percentage of nuclear β-catenin-positive cells were significantly higher than that of the blank control at 3-24 h of DU exposure (p-GSK-3β(S9)/GSK-3β:t=3.95, 4.69, 5.40, 3.34, P<0.05; nuclear β-catenin-positive cells:t=4.61, 6.52, 36.64, 14.93, P<0.05) with a maximum response at 9 h of DU exposure accompanied with corresponding increase of protein level of c-myc, a downstream target gene of β-catenin. Transient transfection of HK-2 cells with GSK-3β(KD) plasmid significantly inhibited the activity of GSK-3β (t=8.07, P<0.05) and reduced the DU-increased percentage of KIM-1-positive cells (t=24.77, P<0.05). Treatment cells with TDZD-8 inhibited the activity of GSK-3β and enhanced the percentage of nuclear β-catenin-positive cells, and it also significantly reduced the percentage of KIM-1-positive cells in HK-2 cells exposed to DU (t=6.25, 6.73, P<0.05). Moreover, overexpression of β-catenin significantly reduced DU-induced cell injury (t=7.48, P<0.05). Conclusions GSK-3β/β-catenin signaling plays a key role in regulating the DU-induced cytotoxicity of HK-2 cells. Inhibition of GSK-3β activity and overexpression of β-catenin can protect the HK-2 cells from DU-induced damage. |
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