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本文分析了人体肝癌细胞 (HepG2 )急性低氧处理以及低氧习服处理后基因表达谱的改变。急性低氧处理为细胞在 1%氧气中培养 48h ,低氧习服处理为细胞在 1%氧气中培养 2 4h ,常氧培养 2 4h ,以此作为一个周期 ,重复 6个周期。联合应用抑制消减杂交技术和cDNA芯片技术 ,筛选HepG2细胞经急性低氧处理与正常培养细胞相比差异表达的基因 ,以及经低氧习服处理细胞与正常培养细胞相比差异表达的基因。结果显示 ,HepG2细胞经急性低氧处理与在常氧条件下培养相比 ,差异表达的基因有 3 7个 ,表达水平全部表现为下调 ,其中包括参与细胞周期、细胞应激、细胞信号转导、细胞骨架形成、转录相关蛋白及细胞代谢相关蛋白的基因 ,1个未知基因序列、4个EST序列、5个线粒体蛋白基因 ,另外有功能不明的蛋白质基因 12个。低氧习服处理的细胞与常氧条件下培养的细胞相比 ,差异表达的基因有 6个 ,其中包括两个线粒体蛋白基因、金属蛋白酶 1基因、转铁蛋白基因、Thymosin .beta 4和TPT1基因。其中线粒体蛋白ND4、转铁蛋白、Thymosin .beta 4和TPT1基因的表达呈上调 ,线粒体ND1及金属蛋白酶1基因的表达水平呈下调。经低氧习服处理后 ,细胞低氧耐受力提高 ,低氧习服处理细胞基因的表达与急性低氧处理细胞和正常培养细胞的基因表达不
This article analyzes the changes of gene expression profiles after acute hypoxic treatment of human hepatocellular carcinoma cells (HepG2) and hypoxia ingestion. Acute hypoxia treatment of cells in 1% oxygen for 48h, hypoxia acclimation treatment of cells cultured in 1% oxygen for 24 hours, nocturnal 2 4h culture, as a cycle, repeat 6 cycles. Combined with suppression subtractive hybridization and cDNA microarray technology, we screened the differentially expressed genes of HepG2 cells compared with the normal cultured cells under acute hypoxia and differentially expressed genes compared with the normal cultured cells. The results showed that 37 differentially expressed genes were down-regulated in HepG2 cells after acute hypoxic treatment compared with that under normoxia. The expression level of HepG2 cells was down-regulated, including cell cycle, cell stress, cell signal transduction , Cytoskeleton formation, transcription related proteins and cell metabolism related genes, one unknown gene sequence, four EST sequences and five mitochondrial protein genes, and another twelve unknown protein genes. There were 6 differentially expressed genes in hypoxia-treated cells compared with those cultured in normoxic conditions, including two mitochondrial protein genes, metalloproteinase 1, transferrin, Thymosin .beta 4 and TPT1 gene. Among them, the expressions of mitochondrial ND4, transferrin, Thymosin .beta 4 and TPT1 were up-regulated while the expression of mitochondrial ND1 and metalloproteinase-1 were down-regulated. Hypobaric hypoxia tolerance increased after hypoxia treatment, and the gene expression of hypoxia-treated cells was not correlated with the gene expression of acute hypoxic-treated cells and normal cultured cells