EGF对食管鳞癌细胞HIF-1α基因表达及功能的影响  

Effect of EGF on the Function and Expression of HIF-1α in Esophageal Squamous Cancer Cells

作　　者：潘琦[1] 施瑞华[1] 朱宏[1] 凌亭生[1] 郝波[1] 

机构地区：[1]南京医科大学第一附属医院消化科,南京210029

出　　处：《临床误诊误治》2013年第12期93-96,共4页Clinical Misdiagnosis & Mistherapy

基　　金：国家自然科学基金青年基金(30800511)

摘　　要：目的研究表皮生长因子(EGF)对人食管鳞癌细胞株HIF-1α基因表达及功能的影响及其对血管生成拟态相关基因表达的影响。方法将人食管鳞癌细胞株Eca-109、TE13分为试验组(加入含EGF 100μg/L的无血清DMEM培养液)和对照组(加入无血清DMEM培养液),分别于常氧及缺氧下培养。应用蛋白质印迹法(Western blot)检测细胞HIF-1α蛋白与血管内皮钙黏附素(sVE-cadherin)、层黏连蛋白(Laminin5γ2)、酪氨酸蛋白激酶受体(EphA2)和基质金属蛋白酶-2(MMP-2)基因蛋白的表达,应用逆转录聚合酶链反应法(RT-PCR)检测HIF-1α与sVE-cadherin、EphA2和MMP-2 RNA的表达,基质胶(Matrigel)三维培养显微镜下观察并计数两组细胞的管状结构。结果常氧和缺氧情况下,与对照组比较,试验组Eca-109、TE13细胞HIF-1α蛋白与sVE-cadherin、Laminin5γ2、EphA2基因蛋白的表达及sVE-cadherin、EphA2 RNA水平的表达均显著增加,差异均有统计学意义(P<0.05);而对MMP-2基因蛋白的表达及RNA水平的表达无明显影响,差异无统计学意义(P>0.05)。Eca-109、TE13细胞均可形成典型的管网状结构,试验组在EGF作用下数目增加,与对照组比较差异有统计学意义(P<0.05)。结论在常氧和缺氧情况下EGF均可促进Eca-109、TE13细胞HIF-1α蛋白与sVE-cadherin、EphA2等基因蛋白及RNA的表达,均可形成典型的管网状结构,且EGF能有效促进肿瘤细胞体外血管生成拟态形成。Objective To study the effect of epidermal growth factor （EGF） on the gene expression and function of HIF-1α in esophagus squamous carcinoma cells and on related gene expression of angiogenesis mimicry.Methods Eca-109 and TE13 cells were divided into EGF group （join serum-free DMEM culture solution with EGF 100 μg/L） and control group （join serum-free DMEM culture solution）,and the two groups were respectively cultured under normoxic and hypoxic conditions.The expressions of HIF-1α,vascular endothelium cadherin （sVE-cadherin）,Laminin5γ2,receptor of tyrosine protein kinase （EphA2） and matrix metalloproteinases-2 （MMP-2） protein were detected with Western blotting,and expressions of HIF-1α,sVE-cadherin,EphA2 and MMP-2 RNA were detected with reverse transcriptase polymerase chain reaction （RT-PCR） method.The capillary structures were observed,and the cell numbers were counted in the two groups under microscope after three dimensional cultivation.Results Compared with control group,the expressions of HIF-1α protein,sVE-cadherin,Laminin5γ2 and EphA2 protein and levels of sVE-cadherin and EphA2 RNA in Eca-109 and TE13 cells under normoxia or under hypoxia conditions in EGF group were significantly increased （P ＜0.05） ; but the differences in expression of MMP2 gene and RNA level showed no statistical significance （P ＞0.05）.Compared with the control group,Eca-109 and TE13 cells could form typical tubuloreticular structure,and the count in EGF group was significantly increased with EGF （P ＜ O.05）.Conclusion EGF could up-regulate the expressions of HIF-1α,sVE-cadherin,EphA2 and RNA in Eca-109 and TE13 cells under normoxia or under hypoxia conditions,and esophageal squamous cancer cell lines Eca-109 and TE13 are all capable of forming typical tubuloreticular structure.EGF can effectively promote their vascularization mimicry formation in vitro.

关 键 词：表皮生长因子 食管肿瘤 缺乏诱导因子 基因表达调控 

分 类 号：R735.1[医药卫生—肿瘤]