ABSTRACT
AIM:To investigate the effect of vitamin D3 up-regulated protein 1 (VDUP1) gene over-expression/knockdown on the proliferation and migration of human breast cancer MCF-7 cells and its related mechanisms.METHODS:Gene over-expression/interference techniques were used to up-regulate/down-regulate the expression of VDUP1 in the MCF-7 cells.The mRNA expression of VDUP1 was detected by qPCR.CCK-8, BrdU and Transwell assays were used to measure the cell viability, proliferation and migration, respectively.The protein levels of Akt, p-Akt, GSK3β and p-GSK3β were determined by Western blot.RESULTS:The mRNA expression of VDUP1 was up-regulated after transfection with VDUP1 over-expression plasmid (P<0.05), and down-regulated after transfection with VDUP1 siRNA (P<0.05).Over-expression of VDUP1 significantly inhibited MCF-7 cell proliferation and migration (P<0.05), while knockdown of VDUP1 enhanced cell proliferation and migration (P<0.05).Furthermore, over-expression of VDUP1 up-regulated the protein levels of p-Akt and p-GSK3β (P<0.05).Inverse results were obtained after knockdown of VDUP1.CONCLUSION:The viability and migration ability of MCF-7 cells are inhibited by over-expression of VDUP1 but enhanced by VDUP1 knockdown, which may be related with Akt/GSK3β pathway.
ABSTRACT
Vitamin D3 up-regulated protein 1 (VDUP1) is a potent growth suppressor that inhibits tumor cell proliferation and cell cycle progression when overexpressed. In a previous study, we showed that VDUP1 knockout (KO) mice exhibited accelerated liver regeneration because such animals could effectively control the expression of cell cycle regulators that drive the G1-to-S phase progression. In the present study, we further investigated the role played by VDUP1 in initial priming of liver regeneration. To accomplish this, VDUP1 KO and wild-type (WT) mice were subjected to 70% partial hepatectomy (PH) and sacrificed at different times after surgery. The hepatic levels of TNF-alpha and IL-6 increased after PH, but there were no significant differences between VDUP1 KO and WT mice. Nuclear factor-kappaB (NF-kappaB), c-Jun-N-terminal kinase (JNK), and signal transducer and activator of transcription 3 (STAT-3) were activated much earlier and to a greater extent in VDUP1 KO mice after PH. A single injection of TNF-alpha or IL-6 caused rapid activation of JNK and STAT-3 expression in both mice, but the responses were stronger and more sustained in VDUP1 KO mice. In conclusion, our findings provide evidence that VDUP1 plays a role in initiation of liver regeneration.
Subject(s)
Animals , Male , Blotting, Western , Carrier Proteins/genetics , Cell Proliferation , Gene Expression Regulation , Hepatectomy , Hepatocytes/cytology , JNK Mitogen-Activated Protein Kinases/genetics , Liver/physiology , Mice, Knockout , NF-kappa B/genetics , Polymerase Chain Reaction , Regeneration , STAT3 Transcription Factor/genetics , Thioredoxins/geneticsABSTRACT
Preadipocyte factor-1 (Pref-1) is expressed in the neuroendocrine organs such as the pituitary gland, the adrenal gland, the pancreas, the testis, etc. Vitamin D3 up-regulated protein 1(VDUP1) gene is known to be a novel member of early response genes as an oxidative stress mediator. The aim of the present study was to investigate whether Pref-1 and VDUP1 is involved in stress response in the adrenal gland following chronic immobilization stress. In situ hybridization for Pref-1 and VDUP1 genes (Pref-1 and VDUP1) was performed in the rat adrenal glands following immobilization stress, 2 hr once daily for 7 days. In situ hybridization analysis revealed that Pref-1 expression was up-regulated in rat adrenal medulla following chronic immobilization stress. However, Pref-1 was down-regulated in the zona glomerulosa of the adrenal cortex following chronic immobilization stress. VDUP1 expression was up-regulated in the zona glomerulosa and the adrenal medulla following chronic immobilization stress. These results show that Pref-1 and VDUP1 may be novel genes responding to chronic immobilization stress in adrenal gland.
Subject(s)
Animals , Rats , Adrenal Cortex , Adrenal Glands , Adrenal Medulla , Cholecalciferol , Immobilization , In Situ Hybridization , Oxidative Stress , Pancreas , Pituitary Gland , Testis , Vitamins , Zona GlomerulosaABSTRACT
AIM: To investigate the expression of vitamin D3 up-regulated protein 1 (VDUP1) in peripheral eosinophils of asthma patients and its relation with eosinophil activation.METHODS: 10 normal volunteers and 31 mild to moderate asthma patients were selected. Symptom severity, pulmonary function index, induced sputum eosinophil counts were recorded. Then, gene and protein expressions of VDUP1 and ?-actin were evaluated by RT-PCR and Western blotting, respectively. In addition, eosinophils were incubated with IL-5, both VDUP1 and ?-actin were amplified by RT-PCR. The eosinophil cationic protein (ECP) of supernatant and serum were also detected by ELISA assay. RESULTS: There was a significant decrease in expression of VDUP1 in asthma attack patients without treatment compared with normal volunteers and patients in remission. In contrast, no significant difference between the patients in remission and normal volunteers was observed. In patients with asthma attacks, a negative relationship between expression intensity of VDUP1 and EOS% in induced sputum and serum ECP concentration was also observed. The expression of VDUP1 in eosinophils was decreased by IL-5 stimulation, simultaneously, the ECP in supernatant was increased. CONCLUSION: The expression of VDUP1 in eosinophils decreases in asthma patients, and is negatively associated with serum ECP and induced sputum EOS%. EOS activation by IL-5 may be related to VDUP1 pathway.