白藜芦醇通过 p53/SLC7A11 对口腔鳞状细胞癌中铁死亡的作用及机制。

Effect and mechanism of resveratrol on ferroptosis mediated by p53/SLC7A11 in oral squamous cell carcinoma.

机构信息

Department of Stomatology, Loudi Central Hospital of Hunan Province, 51 Changqing Middle Street, Loudi, 417000, Hunan, China.

出版信息

BMC Oral Health. 2024 Jul 10;24(1):773. doi: 10.1186/s12903-024-04395-3.

DOI:10.1186/s12903-024-04395-3

PMID:38987730

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11238462/

Abstract

OBJECTIVE

Resveratrol (Res) is a natural phytoestrogen with antitumor activity. This study sought to investigate the role of Res in ferroptosis in oral squamous cell carcinoma (OSCC).

METHODS

Normal human oral keratinocyte (HOK)/oral OSCC (CAL-27/SCC-9) cell lines were treated with different doses of Res. Res toxicity was determined by MTT assay, with half maximal inhibitory concentration values of Res on CAL-27 and SCC-9 cells calculated. Cell viability/colony formation efficiency/migration/invasion/cycle were assessed by CCK-8/colony formation assay/transwell assay/flow cytometry. The expression of p53 protein in the nucleus and cytoplasm, glutathione peroxidase 4 (GPX4) expression, and SLC7A11 messenger RNA (mRNA) and protein expression levels were determined by Western blot and RT-qPCR. Fe content, reactive oxygen species (ROS) level, reduced glutathione (GSH), and lactate dehydrogenase (LDH) release were assessed.

RESULTS

Medium- to low-dose Res had no toxic effect on HOK cells, while high-dose Res markedly reduced HOK cell viability. Res significantly suppressed the viability of OSCC cells (CAL-27 and SCC-9). Res inhibited OSCC cell colony formation/migration/invasion, and induced G1 phase arrest. Res caused the translocation of p53 protein to the nucleus, obviously increased Fe content, ROS level and LDH release, decreased GSH content and GPX4 protein expression, and induced ferroptosis. Down-regulation of p53 partially reversed the inhibitory effects of Res on CAL-27 cell malignant behaviors. Res inhibited SLC7A11 transcription by promoting p53 entry into the nucleus. SLC7A11 overexpression negated the the regulatory effects of p53 knockout on the role of Res in OSCC cell malignant behaviors and ferroptosis.

CONCLUSION

Res accelerated ferroptosis and inhibited malignant behaviors in OSCC cells by regulating p53/SLC7A11.

摘要

目的

白藜芦醇（Res）是一种具有抗肿瘤活性的天然植物雌激素。本研究旨在探讨 Res 在口腔鳞状细胞癌（OSCC）中的铁死亡作用。

方法

用不同剂量的 Res 处理正常人口腔角质细胞（HOK）/口腔 OSCC（CAL-27/SCC-9）细胞系。用 MTT 测定法测定 Res 的毒性，计算 Res 对 CAL-27 和 SCC-9 细胞的半抑制浓度值。用 CCK-8 法/集落形成试验/Transwell 试验/流式细胞术评估细胞活力/集落形成效率/迁移/侵袭/细胞周期。用 Western blot 和 RT-qPCR 检测核和胞质中 p53 蛋白、谷胱甘肽过氧化物酶 4（GPX4）表达以及溶质载体家族 7 成员 11（SLC7A11）信使 RNA（mRNA）和蛋白表达水平。评估铁含量、活性氧（ROS）水平、还原型谷胱甘肽（GSH）和乳酸脱氢酶（LDH）释放。

结果

中低剂量的 Res 对 HOK 细胞没有毒性作用，而高剂量的 Res 则明显降低了 HOK 细胞的活力。Res 显著抑制 OSCC 细胞（CAL-27 和 SCC-9）的活力。Res 抑制 OSCC 细胞集落形成/迁移/侵袭，并诱导 G1 期阻滞。Res 导致 p53 蛋白向核内易位，明显增加铁含量、ROS 水平和 LDH 释放，降低 GSH 含量和 GPX4 蛋白表达，诱导铁死亡。下调 p53 部分逆转了 Res 对 CAL-27 细胞恶性行为的抑制作用。Res 通过促进 p53 进入细胞核来抑制 SLC7A11 的转录。SLC7A11 的过表达否定了 p53 敲除对 Res 在 OSCC 细胞恶性行为和铁死亡中作用的调节作用。