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缺氧诱导因子-1α的抑制影响口腔鳞状细胞癌细胞中自噬介导的糖基化作用。

Inhibition of HIF-1α Affects Autophagy Mediated Glycosylation in Oral Squamous Cell Carcinoma Cells.

作者信息

Li Yi-Ning, Hu Ji-An, Wang Hui-Ming

机构信息

Department of Pathology, Stomatology Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310006, China.

Department of Oral and Maxillofacial Surgery, Stomatology Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310006, China.

出版信息

Dis Markers. 2015;2015:239479. doi: 10.1155/2015/239479. Epub 2015 Nov 11.

Abstract

Purpose. To validate the function of autophagy with the regulation of hypoxia inhibitor-induced glycosylation in oral squamous cell carcinoma cell. Methods. Human Tca8113 cell line was used to detect autophagy and glycosylation related protein expression by western blotting and immunofluorescence with HIF-1α inhibitor. Short interfering RNA (siRNA) transfection blocked human ATG12 and ATG1. Results. HIF-1α inhibitor PX-478 reduced the amount of LC3-II and LC3-I in Tca8113 cells. PX-478 decreased the expression of O-GlcNAc and OGT and increased OGA expression. The tendency of O-GlcNAc showed a similar pattern to OGT. PX-478 gradually decreased OGT expression in Tca8113 cells. Protein level of O-GlcNAc and OGT increased in ATG12 and ATG1 depletion. The expression of OGT decreased at first and then rose slowly with the treatment of Atg12 and Atg1 siRNA and PX-478 fluctuant. Autophagy affected the stability of OGT when HIF-1α signaling was blocked. Conclusions. Autophagy reduced by hypoxic stress inhibited. HIF-1α inhibitor decreased glycosylation. OGT became unstable in the absence of autophagy when HIF-1α signaling was blocked.

摘要

目的。验证自噬在口腔鳞状细胞癌细胞中对缺氧抑制剂诱导的糖基化的调节作用。方法。使用人Tca8113细胞系,通过蛋白质免疫印迹法和免疫荧光法,利用缺氧诱导因子-1α(HIF-1α)抑制剂检测自噬和糖基化相关蛋白的表达。采用小干扰RNA(siRNA)转染阻断人自噬相关基因12(ATG12)和自噬相关基因1(ATG1)。结果。HIF-1α抑制剂PX-478降低了Tca8113细胞中微管相关蛋白1轻链3-II(LC3-II)和微管相关蛋白1轻链3-I(LC3-I)的含量。PX-478降低了O-连接的N-乙酰葡糖胺(O-GlcNAc)和O-连接的N-乙酰葡糖胺转移酶(OGT)的表达,并增加了O-GlcNAc酶(OGA)的表达。O-GlcNAc的变化趋势与OGT相似。PX-478使Tca8113细胞中OGT的表达逐渐降低。在ATG12和ATG1缺失时,O-GlcNAc和OGT的蛋白水平升高。在使用自噬相关基因12(Atg12)和自噬相关基因1(Atg1)的小干扰RNA(siRNA)处理时,OGT的表达先降低,然后缓慢上升,且PX-478的作用呈波动变化。当HIF-1α信号通路被阻断时,自噬影响了OGT的稳定性。结论。缺氧应激导致自噬减少受到抑制。HIF-1α抑制剂降低了糖基化水平。当HIF-1α信号通路被阻断时,在缺乏自噬的情况下OGT变得不稳定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/4658405/c45ba9fb37f8/DM2015-239479.001.jpg

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