摘要
目的:初步探讨p53在低氧抑制人牙周膜成纤维细胞(periodontal ligament cells,PDLCs)增殖、促进其凋亡中的作用。方法:体外正常氧(20%O_2)和低氧(1%O_2)培养PDLCs细胞48 h,Wester n免疫印迹和实时定量PCR检测p53与HIF-1α表达水平;通过小干扰RNA(Si-HIF1α)转染PDLCs,验证敲低HIF-1α表达后p53水平变化;进一步通过小干扰R NA(Si-p53)转染PDLCs,采用四甲基偶氮唑盐(MTT)法检测敲低p53后PDLCs细胞增殖能力差异;流式细胞术比较敲低p53后PDLCs凋亡变化。结果:PDLCs低氧培养48 h后p53与HIF-1α均显著升高,Si-HIF1α转染PDLCs并在低氧培养后HIF-1α表达明显降低,同时伴随p53下降;进一步Si-p53转染PDLCs并于低氧培养后p53水平降低,但HIF-1α表达变化不大,细胞活性显著抑制,凋亡水平增高。结论:低氧微环境中升高的HIF-1α可进一步激活p53表达,进而抑制PDLCs增殖并促进其凋亡。
Objective: To investigate the role of p53 in inhibiting proliferation and promoting apoptosis of human periodontal ligament fibroblasts (periodontal ligament cells, PDLCs) under hypoxia. Methods: PDLCs were cultured under hypoxia (1% O2) or normoxia (20% O2) for 48 h. Western blot and qRT-PCR were used to detect expression of p53 and HIF-1α at protein and mRNA level respectively. Under transfected HIF-1α with siRNA, HIF-1α and p53 expression were furthered detected. After further transfected p53 with siRNA, p53 and HIF-1α expression, cell viability, as well as apoptosis, were furthered detected in PDLCs under hypoxia. Results: The expression of p53 and HIF-1α was significantly increased after PDLCs hypoxia for 48 h, and the expression of HIF-1α was decreased after transfection of Si-HIF1α into PDLCs, which was accompanied by the decrease of p53. Further transfection of PDLCs with Si-p53 decreased the level of p53, but the expression of HIF-1α did not change significantly, the cell activity was significantly inhibited, and the level of apoptosis was increased. Conclusion: Hypoxia increases HIF-1α and p53 expression, and hypoxia inhibits proliferation and promotes apoptosis of PDLCs via upregulated p53.
作者
王勤
庄秀妹
何杰玉
张叶
彭雪珍
WANG Qin;ZHUANG Xiu-mei;HE Jie-yu;ZHANG Ye;PENG Xue-zhen(Department of Stomatology, Shenzhen Children’s Hospital, Shenzhen 518026, China)
出处
《中华老年口腔医学杂志》
2019年第2期70-74,共5页
Chinese Journal of Geriatric Dentistry
基金
国家自然科学基金青年基金项目(项目编号:81600899)
