LncRNA HOTAIR involves in upregulation of HIF-1α in HaCat cells under low oxygen condition / 中华医学美学美容杂志

ABSTRACT

Objective:To observe the effect of LncRNA HOTAIR on the expression of HIF-1α in HaCat cell under low oxygen condition, and to explore the role of LncRNA HOTAIR in the pathogenesis and development of keloid.Methods:From Jan. 2018 to Dec. 2018 in Chinese Academy of Medical Science, recombinant plasmids were designed and constructed by specific shRNA-HOTAIR. After transfected HaCat cells with sh-LncRNA HOTAIR, RT-PCR was used to detect the expression level of HOTAIR. HaCat cells were cultured in different conditions and divided into four groups: group A cultured under normoxia condition, the other three groups cultured under hypoxia condition. Group C was transfected with sh-control before hypoxic culture, while group D were transfected with sh-LncRNA HOTAIR. After 24 hours′ culture, dual luciferase reporter gene assay was used to verify the relationship of LncRNA HOTAIR and HIF-1α. Expression of HIF-1α in four groups of HaCat cells were determined by Western blot analysis.Results:Recombinant plasmids of shRNA-HOTAIR were successfully constructed, and the HOTAIR expression was significantly decreased. The relative luciferase activity was 0.94±0.30 in group A, 20.39±1.15 in group B, 18.09±0.80 in group C and 3.04±1.15 in group D. The relative luciferase activity of group B was higher than those of group A ( t=29.03, P<0.05), and the difference was statistically significant ( P<0.05). According to the results of Western blotting, group B (1.19±0.07) and group D (1.15±0.06) had higher expression of HIF-1α than group A (0.56±0.29) and group C (0.37±0.38). Down-regulation of HOTAIR significantly inhibited the protein level of HIF-1α. Conclusions:LncRNA HOTAIR plays a positive role in upregulation of HIF-1α in HaCat cells under hypoxia condition. Thus LncRNA HOTAIR may take part in the pathogenesis and development of keloid through HIF-1α pathway.