【摘要】 目的:探讨缺氧环境下,卵巢癌细胞株SKOV3中HIF-1α、HIF-2α和Synde-can-1在转录及蛋白水平的表达及相互作用关系。方法:利用MatInspector在线分析软件,分析Syndecan-1的转录因子结合位点。在CoCl2诱导缺氧环境以及低氧诱导因子(HIF)抑制剂雷帕霉素(Rapamycin)的干预下,实时荧光定量RT-PCR检测HIF-1α、HIF-2α及Syndecan-1 mRNA水平的表达;Western blot检测HIF-1α、HIF-2α蛋白水平的表达。结果:Syndecan-1的转录因子结合位点中有3处HIFF(低氧诱导因子家族)/HIF保守序列。缺氧时,HIF-1αmRNA降低(P<0.05),HIF-1α蛋白、HIF-2αmRNA和蛋白、Syndecan-1mRNA水平均升高(P<0.05)。Rapamycin不能抑制CoCl2模拟缺氧条件下HIF-1α的蛋白表达,可抑制HIF-2α的表达。Syndecan-1 mRNA动态变化曲线和HIF-1α及HIF-2α蛋白水平的动态变化曲线一致。结论:低氧可诱导HIF-1α和HIF-2α的表达,并通过HIF-1α、HIF-2α途径增强Syndecan-1的表达。更多还原

【Abstract】 Objective:To investigate the expression of hypoxia inducible factor-1α(HIF-1α),hypoxia inducible factor-2α(HIF-2α),and Syndecans-1,and their relationship during hypoxia in human ovarian cancer cell line SKOV3.Methods:The transcription factor binding sites of Syndecan-1 were analyzed with Matinspector tools.In addition,under hypoxic environments and an exposure to Rapamycin,a suppressor for hypoxia inducible factors,SKOV3 cells were subject to fluorescent quantitative RT-PCR for HIF-1α,HIF-2α and Syndecan-1 mRNA expressions,Western blot was made for HIF-1α and HIF-2α proteins expressions.Results:There were three HIFF(hypoxia-inducible factor family) /HIF conserved sequences in the Syndecan-1 transcription factor binding sites.Under the condition of hypoxia,the expressions of HIF-1α mRNA reduced,the expressions of HIF-1α protein,HIF-2α mRNA and protein increased(P<0.05).Rapamycin couldn’t inhibit the expression of HIF-1α protein,but could inhibit the expression of HIF-2α.The change of Syndecan-1 mRNA was consistent with that of the HIF-1α and HIF-2α proteins.Conclusion:Hypoxia can induce expression of HIF-1α and HIF-2α and promote the expression of Syndecan-1 through the HIF-1α and HIF-2α pathway.更多还原