Discordant mRNA and protein expression of CXCR4 under in vitro CoCl2-induced hypoxic conditions
Discordant mRNA and protein expression of CXCR4 under in vitro CoCl2-induced hypoxic conditions
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体外 CoCl2 诱导的缺氧条件下 CXCR4 mRNA 和蛋白表达不一致
DOI：
10.1016/j.bbrc.2017.01.102
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发表时间：
2017
影响因子：
3.1
通讯作者：
Zhuang Jing
中科院分区：
文献类型：
--
作者：
Tang Mingjun;Yang Ying;Yu Jingzhi;Wu N;an;Chen Pei;Xu Lijun;Wang Qiyun;Xu Zhuojun;Ge Jian;Yu Keming;Zhuang Jing
氯化钴 (CoCl2) 长期以来被认为是一种合适的体外缺氧模拟剂。编码趋化因子受体的基因CXCR4在缺氧性视网膜疾病中发挥着关键作用。在这里，我们通过实时PCR和蛋白质印迹研究了CoCl2诱导的低氧条件下WERI-Rb1视网膜母细胞瘤细胞和人脐静脉内皮细胞（HUVEC）中CXCR4的mRNA和蛋白表达。我们发现，在 12、24、48 和 72 小时时，CoCl2 诱导的缺氧在 mRNA 水平上显着增加了 CXCR4 的表达，但在蛋白质水平上没有增加。有趣的是，这个结果与 1% O2 缺氧条件下的观察结果不同。此外，荧光素酶测定表明，CoCl2 诱导的缺氧显着增加了 CXCR4 启动子的转录。为了比较我们的体外检查结果与体内缺氧的影响，我们构建了 OIR（氧诱导性视网膜病变）大鼠模型。然而，与对照组相比，OIR 大鼠的 CXCR4mRNA 和蛋白质水平均显着增加。综上所述，我们的研究结果表明，在体外 CoCl2 诱导的缺氧条件下，CXCR4mRNA 和蛋白质表达之间的关系并不是严格线性的。通过体外和体内的比较实验，这项研究表明 CoCl2 对视网膜疾病缺氧的模拟并不完美。
Cobalt chloride (CoCl2) has long been accepted as a suitablein vitrohypoxia-mimetic agent. The geneCXCR4, which encodes a chemokine receptor, plays a key role in hypoxic retinal disease. Here, we investigated the mRNA and protein expression ofCXCR4in WERI-Rb1 retinoblastoma cells and human umbilical vein endothelial cells (HUVECs) under CoCl2-induced hypoxic conditions, by means of real-time PCR and western blot. We found that CoCl2-induced hypoxia profoundly increasedCXCR4expression at the mRNA level, but not at the protein level, at 12, 24, 48 and 72 h in these cells. Interestingly, this result differed from observations of 1% O2hypoxic conditions. Additionally, luciferase assays demonstrated that CoCl2-induced hypoxia significantly increased transcription at theCXCR4promoter. In order to compare ourin vitrofindings with the effects of hypoxiain vivo, an OIR (Oxygen-induced retinopathy) rat model was constructed. However, bothCXCR4mRNA and protein levels in OIR rats were significantly increased compared to controls. Thus taken together, our findings suggest that the relationship betweenCXCR4mRNA and protein expression is not strictly linear underin vitroCoCl2-induced hypoxic conditions. through comparativein vitroandin vivoexperiments, this study implies that CoCl2is an imperfect simulation of hypoxia in retinal disease.