具有血小板反应蛋白I型基序的ADAM金属肽酶,具有抗血管生成和聚集蛋白聚糖酶活性的1(ADAMTS1)在许多病理生理情况下失调。然而,关于ADAMTS1基因转录调控的信息有限。因此,本研究的目的是确定在常氧和低氧条件下对人肝癌细胞(HEP3B)ADAMTS1基因调控的重要调控区。培养的HEP3B细胞暴露于正常的氧气条件下,和氯化钴(CoCl2)诱导的缺氧条件,是HIF-1诱导物。cocl2诱导的缺氧条件导致肝癌细胞诱导ADAMTS1mRNA和蛋白表达。通过转录活性测定SP1和USF转录因子对ADAMTS1基因表达的差异调控,ADAMTS1基因的mRNA和蛋白水平。SP1和USF转录因子的异位表达导致所有启动子构建体的ADAMTS1转录活性降低,与常氧条件下的mRNA和蛋白质水平一致。然而,在低氧条件下,SP1和USF的过表达导致ADAMTS1基因在mRNA和蛋白水平的表达增加。另一方面,C/EBPα转录因子对ADAMTS1基因mRNA表达无统计学意义,常氧和低氧条件下的蛋白质和转录水平。
ADAM metallopeptidase with thrombospondin type I motif, 1 (ADAMTS1) that has both antiangiogenic and aggrecanase activity was dysregulated in many pathophysiologic circumstances. However, there is limited information available on the transcriptional regulation of ADAMTS1 gene. Therefore, this study mainly aimed to identify regulatory regions important for the regulation of ADAMTS1 gene under normoxic and hypoxic conditions in human hepatoma cells (HEP3B). Cultured HEP3B cells were exposed to normal oxygen condition, and Cobalt chloride (CoCl2) induced the hypoxic condition, which is an HIF-1 inducer. The cocl2-induced hypoxic condition led to the induced ADAMTS1 mRNA and protein expression in Hepatoma cells. Differential regulation of SP1 and
USF transcription factors on ADAMTS1 gene expression was determined by transcriptional activity, mRNA and protein level of ADAMTS1 gene. Ectopic expression of SP1 and
USF transcription factors resulted in the decrease in ADAMTS1 transcriptional activity of all promoter constructs consistent with mRNA and protein level in normoxic condition. However, overexpression of SP1 and
USF led to the increase of ADAMTS1 gene expressions at mRNA and protein level in hypoxic condition. On the other hand, C/EBPα transcription factor didn\'t show any statistically significant effect on ADAMTS1 gene expression at mRNA, protein and transcriptional level under normoxic and hypoxic condition.