Effect of hypoxia on the proliferation and expressions of hypoxia-inducible factor-1α, vascular endothelial growth factor and matrix metalloproteinase-9 in keratinocytes obtained from oral lichen planus lesions
10.3760/cma.j.issn.1002-0098.2015.02.006
- VernacularTitle:缺氧对口腔扁平苔藓角质形成细胞增殖及缺氧诱导因子1α、血管内皮生长因子、基质金属蛋白酶9表达的影响
- Author:
Xiaxia WANG
1
;
Guoyao TANG
;
Hongying SUN
Author Information
1. 复旦大学附属华山医院口腔科
- Keywords:
Lichen planus,oral;
Hypoxia-inducible factor 1,alpha subunit;
Matrix metalloproteinase 9;
Keratinocytes
- From:
Chinese Journal of Stomatology
2015;50(2):89-94
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of hypoxia on the proliferation and expressions of hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor(VEGF) and matrix metalloproteinase-9(MMP-9) in keratinocytes obtained from oral lichen planus(OLP) lesions.Methods Hypoxia environment was induced by a airtight incubator.Five groups were included,normoxia control group,hypoxia control group(12,24,36,48 h).The effect of different treatment time of hypoxia on cellular proliferation was determined with cell counting kit-8(CCK-8).The mRNA and protein expressions of HIF-lα,VEGF and MMP-9 were analyzed respectively by quantitative real-time polymerase chain reaction with SYBR Green Ⅰ and Western blotting.Results The growth activity of keratinocytes obtained from OLP lesions in the hypoxia groups(0.340±0.002,0.415±0.006,0.546±0.006) was reduced than that in control (0.431±0.001,0.620±0.004,1.022±0.005)(P＜0.01).The mRNA levels of VEGF(2.087±0.291,3.189±0.573,5.402±0.563) and MMP-9(2.936±0.500,4.083±0.300,6.374±0.858) were elevated by hypoxia(P＜0.05).The protein levels of HIF-lα(0.414±0.093,0.751 ±0.056,0.875±0.040),VEGF(0.393 ±0.046,0.557±0.078,0.767±0.045) and MMP-9(0.250±0.053,0.384±0.038,0.611±0.092) were all remarkably elevated by hypoxia (P＜O.05).However,hypoxia had no effect on HIF-1α mRNA expression.The mRNA expression of HIF-1α after hypoxia exposure for 36 h(1.412±0.094) and 48 h(1.417±0.446) was higher than that of control group,however,there was no significant difference.A positive correlation was noted between HIF-1α and VEGF in protein level(r=0.905,P=O.O00),and the same correlation found between HIF-1α and MMP-9(r=0.881,P=0.000).Conclusions Hypoxia conditions may inhibit the proliferation of keratinocytes obtained from OLP lesions.Hypoxia conditions can promote the protein expressions of HIF-1α and both the mRNA and protein expression of VEGF and MMP-9 in keratinocytes obtained from OLP lesions exposed to hypoxia.The relative high expression of HIF-lα may be involved in multiple aspects of OLP progression through the regulation of its downstream target genes.
