褪黑素通过抑制间充质样 CD44 OSCC 细胞中的 DERL1 增强顺铂诱导的细胞死亡。

Melatonin enhances cisplatin-induced cell death through inhibition of DERL1 in mesenchymal-like CD44 OSCC cells.

机构信息

Department of Public Oral Health, Program of Oral Health Sciences, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

Department of Oral and Maxillofacial Surgery, Program of Dentistry, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

出版信息

J Oral Pathol Med. 2022 Mar;51(3):281-289. doi: 10.1111/jop.13242. Epub 2021 Oct 17.

DOI:10.1111/jop.13242

PMID:34551150

Abstract

BACKGROUND

Melatonin is a hormone that is primarily produced in the pineal gland and is involved in wide range of biological functions. However, the impact of melatonin on chemotherapy-induced cell death remains to be elucidated in oral squamous cell carcinoma (OSCC) cells. The objective of this study was to clarify the role of melatonin in cisplatin-induced cytotoxicity in CD44 OSCC cells.

METHODS

CD44 OSCC cells were cultured on fibronectin-coated hydrogel. A lactate dehydrogenase cytotoxicity assay was performed to evaluate cisplatin-induced cell death. The effect of melatonin on cisplatin-induced cell death and Derlin-1 (DERL1) endoplasmic reticulum membrane protein expression was investigated.

RESULTS

CD44 OSCC cells exhibited mesenchymal-like features when cultured on fibronectin-coated hydrogel. Mesenchymal-like CD44 OSCC cells demonstrated strong resistance to cisplatin-induced cell death compared with epithelial-like CD44 OSCC cells. DERL1 mRNA and DERL1 protein expression levels were significantly higher in mesenchymal-like CD44 cells compared with epithelial-like CD44 cells. Cisplatin-induced cell death was significantly enhanced after DERL1 siRNA knockdown, suggesting that DERL1 is involved in resistance to cisplatin-induced cell death. Melatonin significantly inhibited DERL1 expression and enhanced cisplatin-induced cell death in mesenchymal-like CD44 cells. miR-181c-5p expression was significantly upregulated in the presence of melatonin. Furthermore, melatonin-inhibited DERL1 expression was significantly recovered by miR-181c-5p inhibitor. In addition, melatoninenhanced cisplatin-induced cell death was attenuated by miR-181c-5p inhibitor. These results suggest that melatonin-induced miR-181c-5p enhances cisplatin-induced cell death through inhibition of DERL1 in mesenchymal-like CD44 cells.

CONCLUSIONS

Melatonin plays a vital role in promoting cisplatin-induced cytotoxicity in mesenchymal-like CD44 OSCC cells.

摘要

背景

褪黑素是一种主要在松果体中产生的激素，参与广泛的生物学功能。然而，褪黑素对口腔鳞状细胞癌（OSCC）细胞中化疗诱导的细胞死亡的影响仍需阐明。本研究旨在阐明褪黑素在 CD44 OSCC 细胞中顺铂诱导细胞毒性中的作用。

方法

将 CD44 OSCC 细胞在纤维连接蛋白包被的水凝胶上培养。通过乳酸脱氢酶细胞毒性测定法评估顺铂诱导的细胞死亡。研究了褪黑素对顺铂诱导的细胞死亡和 Derlin-1（DERL1）内质网膜蛋白表达的影响。

结果

CD44 OSCC 细胞在纤维连接蛋白包被的水凝胶上培养时表现出间充质样特征。与上皮样 CD44 OSCC 细胞相比，间充质样 CD44 OSCC 细胞对顺铂诱导的细胞死亡具有更强的抗性。与上皮样 CD44 细胞相比，间充质样 CD44 细胞中 DERL1 mRNA 和 DERL1 蛋白表达水平显著升高。DERL1 siRNA 敲低后，顺铂诱导的细胞死亡明显增强，表明 DERL1 参与了顺铂诱导的细胞死亡抵抗。褪黑素显著抑制间充质样 CD44 细胞中 DERL1 的表达，并增强顺铂诱导的细胞死亡。存在褪黑素时，miR-181c-5p 的表达显著上调。此外，褪黑素抑制的 DERL1 表达通过 miR-181c-5p 抑制剂显著恢复。此外，miR-181c-5p 抑制剂减弱了褪黑素增强的顺铂诱导的细胞死亡。这些结果表明，褪黑素诱导的 miR-181c-5p 通过抑制间充质样 CD44 细胞中的 DERL1 增强顺铂诱导的细胞死亡。