低氧应激对人肝癌细胞AFP、VEGF、TIMP-1、MMP-9表达的影响.pdfVIP

第 42卷第 1期 温 州 医 学 院 学 报 VO1．42NO．1 20l2年 1月 JournalofWenzhouMedicalCollege Jan．2012 蠹 低氧应激对人肝癌细胞AFP、VEGF、TIMP一1、 MMP一9表达的影响 吴之 浩 (义乌市中心医院 普外一科，浙江 金华 322000) [摘 要] 目的：探讨低氧应激对人肝癌细胞 (HepG一2细胞)甲胎蛋白(AFP)、血管内皮生长因子 (VEGF)、 金属蛋 白酶组织抑制剂 (TIMP)一1、基质金属蛋 白酶 (MMP)一9表达的影响。方法：低氧 (5％0、5％CO。、 90％N)和常氧培养HepG一2细胞。逆转录一多聚酶链式反应技术测定HepG一2细胞AFP、VEGF、TIMP—l、MMP一 9mRNA的表达。酶联免疫吸附法测定HepG一2细胞培养上清VEGF、TIMP—l、MMP一9蛋 白含量，放射免疫测定 法测定HepG一2细胞培养上清AFP蛋白含量。结果：低氧组AFP、VEGF、MMP一9mRNA和蛋白表达量显著高于 常氧组；TIMP—lmRNA和蛋白表达量显著低于常氧组 (尸0．O1)。结论：低氧应激上调人HepG一2细胞AFP、 VEGF、MMP一9基因表达，下调TIMP一1基因表达。 [关键词] 低氧应激；甲胎蛋白；血管内皮生长因子；金属蛋白酶组织抑制剂一l；基质金属蛋白酶一9 [中图分类号] R364．4；R735．7 [文献标志码] A [文章编号] 1000-2138(2012)01-0041—03 Effects of hypoxia stress on expressions of AFP，VEGF，TIMP-I and MMP-9 in HepG-2 cell ∥∥ Zhihao． TheF1，stDepartment of General Surgery． Y1wu Central Hospital。 Jinhua． 322000 ObJective：Toinvestigatewhetherexpressionsofalpha—fetoprotein (AFP)， vascularendothelialgrowthfactor (VEGF)，tissueinhibitorofmetalloproteinase (TIMP)-1 andmatrixmetalloproteinase(MMF)一9 inHepG一2cellareregulatedbyhypoxia stressornot． Methods：HepG一2cellswere culturedunderhypoxia(5％0，5％cO。and90％N)ornormoxia．The expressions of AFP，VEGF，TIMP—l and MMP-9 mRNA in HepG一2 cel1 were detected with Feverse transcription polymerase chain reaction (RT—PCR)．The expressionsofVEGF，TIMP-1，MMP-9 protein in supernatant of HepG一2 cell were studied with enzyme一1abeled immunosorbent assay (ELISA)．TheexpressionofAFPprotein insupernatantofHepG一2ce11wasdeterminedwith radioimmunityassay(RIA)．Results：TheexpressionsofAFP，VEGFandMMP一9inhypoxiagroup were dramatically higher than those in normoxia group， while the expression of TIMP-1 in hypoxia group was significantly lower than that in normoxia group (P0．01)．Conclusion： The expressions of AFP，VEGF and MMP一9 gene in HepG一2