低氧及HIF-1α基因转染对HepG2细胞基因表达谱的影响

Effects of hypoxia and HIF-1α gene transfection on gene expression profiles of HepG2 cells

目的:应用基因芯片技术,研究缺氧及缺氧诱导因子1α(HIF-1α)基因转染对HepG2细胞基因表达谱的影响。方法:以常氧对照腺病毒(Ad-GFP)转染组(CC组)为对照组,常氧HIF-1α腺病毒(Ad-HIF)转染组(CH组)、低氧对照腺病毒(Ad-GFP)转染组(HC组)、低氧(2%O2,24 h)HIF-1α腺病毒(Ad-HIF)转染组(HH组)为实验组,提取HepG2细胞的总RNA,反转录成Cy3和Cy5标记的cDNA探针,与定制的包含1 628个能量代谢相关基因的cDNA芯片进行杂交,筛选出实验组与对照组差异表达基因并进行分析。结果:与常氧对照腺病毒转染组相比较,常氧HIF-1α腺病毒转染组3个基因表达下调;低氧对照腺病毒转染组10个基因表达上调;低氧HIF-1α腺病毒转染组6个基因表达上调、1个基因表达下调。这些差异表达基因编码蛋白的功能涉及能量代谢、信号转导、细胞凋亡、转录和HIF-1调节、生物转化、酸碱平衡调节、细胞黏附分子等方面。结论:低氧可以上调HepG2细胞能量代谢相关基因表达,HIF-1的转录调节作用可能是其重要环节。

AIM： To investigate the effects of hypoxia and hypoxia - inducible factor -1α （ HIF-1α） gene transfection on gene expression profiles in human HepG2 cells using microarray technique. METHODS： HepG2 cells were divided into four groups ： normoxia with Ad - GFP infection group （ CC）, normoxia with Ad - HIF -1α infection group （CH）, hypoxia （2% O2, 24 h） with Ad- GFP infection group （HC）, hypoxia （2% O2, 24 h） with Ad - HIF- lot infection group （HH）. The mRNA was isolated, reversely transcribed into cDNAs and marked with Cy3 and Cy5. The cDNAs were subjected for microarray screening with 1628 cDNA probes. RESULTS ： Compared to normoxia with Ad - GFP infection group, 10 up -regulated genes were found in hypoxia with Ad -GFP infection group, 6 up -regulated genes and 1 down regulated genes were found in hypoxia with Ad - HIF - lot infection group, and 3 down -regulated genes were found in normoxia with Ad- HIF- lot infection group, respectively. These different expressions of genes were involved in energy metabolism, signal transduction, apoptosis, transcription factors, biological transformation, acid - base balance and adherence molecule. CONCLUSION： Hypoxia is able to up - regulate the expression of metabolic genes in HepG2 cells via transcriptional response controlled by HIF- 1.