HIF-1α/MMP9信号通路介导低氧诱导的人视网膜母细胞瘤HXO-RB44细胞侵袭  

Hypoxia Enhances the Invasion of Retinoblastoma HXO-RB44 Cell Line in Vitro through HIF-1α/MMP9 Pathway

作　　者：荣媛 刘明华 邓朝霞 任小宝 向强 RONG Yuan;LIU Ming-hua;DENG Zhao-xia;REN Xiao-bao;XIANG Qiang(Department of emergency,Southwest Hospital in Third Military Medical University,Chongqing,400038,China)

机构地区：[1]解放军陆军军医大学西南医院急诊科

出　　处：《现代生物医学进展》2019年第11期2036-2040,2018,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81071537)

摘　　要：目的:探究低氧条件下人视网膜母细胞瘤细胞侵袭能力的变化及其调控机制。方法:将人视网膜母细胞瘤HXO-RB44细胞株于常氧(21%O2)和低氧(1%O2)条件下培养,采用Transwell侵袭实验检测细胞侵袭能力,real time-PCR和Western blot法检测细胞中HIF-1α及MMP9 m RNA和蛋白的表达水平,荧光素酶报告基因分别检测低氧对HIF-1α、HIF-1α对MMP9启动子区域的调控作用。并进一步使用HIF-1α及MMP9特异性si RNA阻断其表达,并检测低氧对细胞侵袭能力的影响。结果:低氧组Transwell下室HXO-RB44每高倍视野细胞数目、HXO-RB44HIF-1α和MMP9 m RNA和蛋白表达均较常氧组显著上调(P<0.05)。转染HIF-1α过表达质粒组的MMP9活性较空载组相比明显增加(P<0.05),低氧处理24小时的HXO-RB44细胞的HIF-1α启动子活性较常氧组明显增强(P<0.05)。HIF-1αsi RNA组的HIF-1α及MMP9的m RNA和蛋白表达水平较NC组明显降低(P<0.05),MMP9 si RNA组的HIF-1αm RNA和蛋白无明显改变,分别转染HIF-1αsi RNA组MMP9 si RNA组下室HXO-RB44每高倍视野细胞数目显著减少(P<0.05)。结论:低氧能够促进HXO-RB44细胞株的侵袭转移,而可HIF-1α/MMP9轴在这一过程起关键作用。Objective: To investigate the effect of hypoxia on the invasion ability of retinoblastoma cell and its underlying mechanism.Methods: HXO-RB44 cells were cultured under normoxia(21% O2) and hypoxia(1% O2) conditions for 24 hours. The effect of hypoxia on invasion ability of HXO-RB44 cells were monitored with transwell invasion assay;the m RNA and protein expression levels of HIF-1αand MMP9 were detected by RT-PCR and western blot;luciferase assay was performed to assess the HIF-1α regulation by hy-poxia, and MMP9 regulation by HIF-1α. Furthermore, HIF-1α and MMP9 si RNA was used to investigate the effect of HIF-1α/MMP9 signaling on hypoxia induced cell invasion. Results: The invaded cell number, m RNA and protein expression of HIF-1α and MMP9 in hypoxia group were higher than those in the normoxia group(P<0.05). The activity of MMP9 in HIF-1α overexpression group was higher than that of the vector group(P<0.05). Compared with the normoxia group, the activity of HIF-1α in hypoxia group was signifi-cantly elevated(P<0.05). Compared with negative control group, the m RNA and protein expression of HIF-1α and MMP9 in HIF-1α si RNA group were remarkedly decreased(P<0.05), whereas the m RNA and protein expression of HIF-1α in MMP9 si RNA group didn’t change. The invaded cells in HIF-1α si RNA group and MMP9 si RNA group were significantly decreased(P<0.05). Conclusion: Hypoxia promotes the invasion of retinoblastoma cell line HXO-RB44 and the activation of HIF-1α/MMP9 signaling pathway is in-volved in this process.

关 键 词：低氧 低氧诱导因子-1α 基质金属蛋白酶-9 侵袭 视网膜母细胞瘤 

分 类 号：R-33[医药卫生] R739.7