Beijing Institute of Dental Research, Beijing Key Laboratory, Beijing Stomatological Hospital and School of Stomatology, Capital Medical University, Dongcheng District, Beijing, China.
Beijing Institute of Dental Research, Beijing Key Laboratory, Beijing Stomatological Hospital and School of Stomatology, Capital Medical University, Dongcheng District, Beijing, China.
Arch Oral Biol. 2022 Sep;141:105491. doi: 10.1016/j.archoralbio.2022.105491. Epub 2022 Jun 18.
As a major risk factor for oral leukoplakia (OLK), oxidative stress can induce intracellular reactive oxygen species (ROS), which is closely related to autophagy. Ras-related protein 7 (Rab7) is an important molecule involved in autophagy. Peroxiredoxin 1 (Prx1) is a key antioxidant protein, overexpressed in a variety of malignant tumors. We found that the expression of Prx1 in OLK was up-regulated, and Prx1 is associated with autophagy. This study aims to identify mechanisms of Prx1 in oxidative stress associated autophagy in human dysplastic oral keratinocyte (DOK) cells.
Hydrogen peroxide (HO) was used to induce autophagy in DOK cells. CCK8 assay and flow cytometry were conducted to examine cell viability, cell apoptosis and intracellular ROS level. Autophagy-associated proteins were detected by western blot and immunofluorescence. MHY1485 was applied to investigate the role of Prx1 in autophagy. On the basis of online docking tool Zdock, Prx1 interacting with Rab7 was verified by immunofluorescence double-staining, Duolink PLA, and Co-immunoprecipitation (Co-IP).
HO induced autophagy and ROS increase in DOK cells, and the expression of Prx1 increased gradually according to HO concentration Prx1 knockdown attenuated the inhibition of MHY1485 on the expressions of autophagy-related proteins and the ratio of LC3 II/LC3 I induced by HO. Moreover, Prx1 interacted with Rab7 in DOK cells.
Prx1 was involved in the regulation of HO-induced autophagy in DOK cells partly by physically interacting with Rab7. Prx1 knockdown promoted autophagy maybe by releasing more Rab7 into the autophagy process.
氧化应激作为口腔白斑（OLK）的主要危险因素，可诱导细胞内活性氧（ROS）产生，这与自噬密切相关。Ras 相关蛋白 7（Rab7）是自噬过程中的重要分子。过氧化物还原酶 1（Prx1）是一种关键的抗氧化蛋白，在多种恶性肿瘤中过表达。我们发现 Prx1 在 OLK 中的表达上调，并且 Prx1 与自噬有关。本研究旨在鉴定 Prx1 在人异型口腔角质形成细胞（DOK）细胞氧化应激相关自噬中的作用机制。
过氧化氢（HO）用于诱导 DOK 细胞自噬。CCK8 检测和流式细胞术用于检测细胞活力、细胞凋亡和细胞内 ROS 水平。通过 Western blot 和免疫荧光检测自噬相关蛋白。MHY1485 用于研究 Prx1 在自噬中的作用。基于在线对接工具 Zdock，通过免疫荧光双染、Duolink PLA 和 Co-IP 验证 Prx1 与 Rab7 的相互作用。
HO 诱导 DOK 细胞自噬和 ROS 增加，且随着 HO 浓度的增加，Prx1 的表达逐渐增加。Prx1 敲低减弱了 MHY1485 对 HO 诱导的自噬相关蛋白表达和 LC3 II/LC3 I 比值的抑制作用。此外，Prx1 在 DOK 细胞中与 Rab7 相互作用。
Prx1 部分通过与 Rab7 物理相互作用参与调节 DOK 细胞中 HO 诱导的自噬。Prx1 敲低可能通过将更多的 Rab7 释放到自噬过程中促进自噬。
