| Objective:Urotensin II (UII) was originaly isolated from the urophysis, a neurosecretory organ on the caudal portion of spinal cord of fish called teleost. It's a cyclic peptide and G-protein coupled receptor 14 is the specific receptor. Up to now, UII is the most potent vasoconstrictor in the mammals. To investigate the role of UII and its receptor, GPR14, in the pathogenesis of human pheochromocytoma, we studied the effects of UII on the proliferation of rat pheochromocytoma cell line (PC 12) and primary culture of human pheochromocytoma cells in vitro; we also measured the expression of UII and GPR14 mRNA in normal adrenal cortex and medulla, and human pheochromocytoma tissues.Methods:1. We observed the effects of UII at different concentrations (10-10, 10-9, 10-8, 10-7, 10-6M) on the proliferation of rat PC12 cells with MTT method, and then, PC12 cells were stimulated in the presence of 10-7M UII, whose effects on the proliferation of PC12 cells was observed at 12, 24, 36,48, 60, 72 hr.2. The human pheochromocytoma tissue was digested consecutively for 2-3 times with collagenase type IA, and freed tumor cells were separated for primary culture. MTT was applied to observe the effects of UII at different concentrations (10-10, 10-9, 10-8, 10-7, 10-6M) on the proliferation of human pheochromocytoma cells.3. Total RNA from normal adrenal and tumoral tissues was extracted. After reverse transcription, UII and its internal control GAPDH, and GPR14 and its internal control B-actin, were co-amplified with PCR. Semi-quantitative analysis of the mRNA expression level of UII and GPR14 was carried out based on the ratios of... |
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