Tribbles homolog 3 is induced by high glucose and associated with apoptosis in human endothelial cells
| Autor: | Zhong‑Xiu Guo, Yun Zhang, Ming Zhong, Ling Guo, Wei Zhang, Hui‑Ping Gong, Yuan‑Yuan Shang |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Cancer Research
Apoptosis Cell Cycle Proteins Protein Serine-Threonine Kinases Biology Biochemistry Umbilical vein Downregulation and upregulation Western blot Human Umbilical Vein Endothelial Cells Genetics medicine Humans RNA Messenger RNA Small Interfering Molecular Biology Gene knockdown medicine.diagnostic_test Transfection Molecular biology Repressor Proteins Glucose Real-time polymerase chain reaction Gene Expression Regulation Oncology TRIB3 Molecular Medicine RNA Interference |
| Zdroj: | Molecular Medicine Reports. 12:1963-1970 |
| ISSN: | 1791-3004 1791-2997 |
| DOI: | 10.3892/mmr.2015.3576 |
| Popis: | Tribbles homolog 3 (TRIB3) is an intracellular kinase-like molecule that modifies cellular survival and metabolism. The present study aimed to investigate the function of TRIB3 regulation in the process of high glucose-induced apoptosis in endothelial cells, with the aim of identifying a novel intervention target for the prevention and treatment of diabetes mellitus. Human umbilical vein endothelial cells (HUVECs) grown in medium with various concentrations of glucose (5.5, 10, 20, 30 and 40 mmol/l) were assessed for mRNA expression of TRIB1, TRIB2 and TRIB3 using reverse transcription quantitative polymerase chain reaction. In addition, protein expression of TRIB3 was examined using western blot analysis. Immunofluorescence staining was performed in order to determine the distribution and localization of TRIB3 in HUVECs. Furthermore, cells grown in normal (5.5 mmol/l) or high glucose (HG; 30 mmol/l) medium were subjected to TRIB3 inhibition through small interfering (si)RNA knockdown. These cells were then examined in order to determine whether TRIB3 upregulation was associated with endothelial cell apoptosis. HUVECs treated with 30 and 40 mmol/l glucose for 48 h and 72 h showed significantly lower survival rates compared with those treated with normal glucose levels. In addition, slight but not significant increases in TRIB1 and TRIB2 mRNA expression were observed in HUVECs incubated with various concentrations of glucose for different durations. By contrast, TRIB3 mRNA expression was increased 7.2-fold following incubation with HG. Western blot analysis revealed a 5.44-fold increase in TRIB3 protein levels in cells grown in HG medium for 24 h compared with those grown in normal medium. Immunostaining assays revealed a markedly higher and well-defined nucleolar fluorescence intensity for TRIB3 expression at 24 h in HG medium compared with that of the control group. Furthermore, the apoptotic rate of HG-treated TRIB3 siRNA-transfected HUVECs was significantly increased compared with that of those transfected with control siRNA In conclusion, the results of the present study suggested that TRIB3 was associated with high glucose-induced HUVECs apoptosis, which was attenuated following transfection with TRIB3 siRNA. |
| Databáze: | OpenAIRE |
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