Department of Radiation Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangdong, China.
Jiangmen Central Hospital, Affiliated Jiangmen Hospital of Sun Yat-sen University, Guangdong, China.
J Dermatol Sci. 2020 Oct;100(1):67-74. doi: 10.1016/j.jdermsci.2020.08.012. Epub 2020 Sep 1.
Human Apurinic/Apyrimidinic Endonuclease 1 (APE1/REF-1/HAP1) is a multifunction protein involved in the progression of cancer. But the role of APE1 in cutaneous squamous cell carcinoma (cSCC) is unclear.
This study is aimed to investigate the basic modulatory mechanism of APE1 in cSCC development and offer a novel potential target for clinical treatment.
The expression of APE1 in cSCC tissues was detected by western blot and immunohistochemistry (IHC) staining. The function of APE1 and miR-27a in cSCC cells was investigated by cell counting kit-8 (CCK-8) assays, colony formation assays and transwell migration assays. Western blot was used to determine the expression of APE1 in cSCC and epithelial-mesenchymal transition (EMT) markers in HSC-1 and HSC-5 cells with APE1 knockdown or overexpression. Double luciferase reporter assays were performed to confirm the interaction of miR-27a and APE1.
We identified that APE1 was significantly upregulated in human cSCC tissues and cSCC cells and its overexpression promoted cell proliferation, migration and the expression of EMT markers in cSCC cells. Mechanistically, miR-27a was predicted and confirmed as the upstream of APE1. Its downregulation also enhanced the proliferation and migration of cSCC cells. Rescue experiments demonstrated that restoration of APE1 expression significantly abolished the inhibition of cell proliferation and migration mediated by miR-27a.
As a direct gene of miR-27a, APE1 improved cell proliferation and migration to promote the progression of cSCC, which could be considered as a potential therapeutic target for cSCC treatment.
人类脱嘌呤/脱嘧啶核酸内切酶 1（APE1/REF-1/HAP1）是一种多功能蛋白，参与癌症的进展。但是，APE1 在皮肤鳞状细胞癌（cSCC）中的作用尚不清楚。
本研究旨在探讨 APE1 在 cSCC 发生发展中的基本调节机制，并为临床治疗提供新的潜在靶点。
通过 Western blot 和免疫组织化学（IHC）染色检测 cSCC 组织中 APE1 的表达。通过细胞计数试剂盒-8（CCK-8）检测、集落形成实验和 Transwell 迁移实验研究 APE1 和 miR-27a 在 cSCC 细胞中的功能。Western blot 用于检测 APE1 在 APE1 敲低或过表达的 HSC-1 和 HSC-5 细胞中 cSCC 和上皮-间充质转化（EMT）标志物的表达。双荧光素酶报告基因实验证实 miR-27a 与 APE1 的相互作用。
我们发现 APE1 在人 cSCC 组织和 cSCC 细胞中显著上调，其过表达促进了 cSCC 细胞的增殖、迁移和 EMT 标志物的表达。机制上，miR-27a 被预测并确认为 APE1 的上游分子。其下调也增强了 cSCC 细胞的增殖和迁移。挽救实验表明，APE1 表达的恢复显著消除了 miR-27a 介导的细胞增殖和迁移抑制。
作为 miR-27a 的直接靶基因，APE1 促进了 cSCC 细胞的增殖和迁移，从而促进了 cSCC 的进展，可被视为治疗 cSCC 的潜在靶点。
