Effects of acute hypoxia and intermittent hypoxic acclimatization on vascular endothelial growth factor gene expression in HepG2 cells / 中国应用生理学杂志

<p><b>AIM</b>To explore the influence of acute hypoxia and intermittent hypoxic acclimatization on vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1alpha (HIF-1alpha) gene expression in HepG2 cells underlying their possible biological significance.</p><p><b>METHODS</b>HepG2 was cultured in 1% O2 for 24 hours, then in 21% O2 for another 24 hours, which composed a hypoxic exposure cycle. After 6 cycles, HepG2 cells reached the status of hypoxic acclimatization. Gene transcription and translation of VEGF and HIF-1alpha were detected with Northern blot and Western blot methods.</p><p><b>RESULTS</b>Acute hypoxia could induce gene transcription and translation of VEGF and HIF-1alpha. After intermittent hypoxia acclimatization, the contents of VEGF and HIF-1alpha mRNA were 108.6% +/- 17.7% and 116.7% +/- 19.8% of those in normoxic control cells, while the protein contents were significantly increased to 1.4 and 2.7 times of those in control cells, respectively (P < 0.05). The protein expression levels of VEGF and HIF-1alpha were decreased in cells subjected to hypoxia acclimatization compared to cells treated with acute hypoxia.</p><p><b>CONCLUSION</b>When HepG2 cells reached the status of hypoxic acclimatization, the acute hypoxia-induced increment of VEGF gene transcription and translation in cells were inhibited, in which HIF-1alpha might play an important role.</p>

Identification of differentially expressed genes of acute hypoxia-treated HepG2 cells and hypoxia-acclimatized HepG2 cells / 生理学报

To provide necessary information for further understanding of molecular mechanism of hypoxia acclimatization, the differentially expressed genes of HepG2 cells exposed to normoxia, acute hypoxia-treated cells which were exposed to 1% oxygen for 48 h, and hypoxia-acclimatized HepG2 cells which were cultured for 6 circles of alternate low oxygen (1% oxygen for 24 h) and normal oxygen (21% oxygen for 24 h), were identified respectively by combining the suppression subtractive hybridization (SSH) and cDNA microarray. Thirty-seven genes were expressed differentially in cells exposed to 1% oxygen for 48 h compared with those in cells exposed to normoxia. The expression of all these 37 genes was down-regulated, including the genes participating in cell cycle, cell response to stimulus, and cell signal transduction, and cell cytoskeleton formation, the genes associated with transcription and cell metabolism, 4 expressed sequence tags (ESTs), and 12 genes of which the functions are not known. There is a novel gene sequence, which has not been found in existing databases. There were only 6 genes differentially expressed in the hypoxia-acclimatized cells compared with cells exposed to normoxia, including two mitochondrion genes, metalloprotease-1 gene, ferritin gene, thymosin beta-4 and TPT1 genes. The expressions of mitochondrion ND4, ferritin, thymosin beta-4 and TPT1 were up-regulated, while the expressions of mitochondrion ND1 gene and metalloproease-1 gene were down-regulated. Cell tolerance to hypoxia increased after the cells were hypoxia-acclimatized. The different gene expression patterns of the acute hypoxia-treated cells and the hypoxia-acclimatized cells may be related to the increased tolerance of the cells to hypoxia.