Liu Yujia, Zhang Yiwen, Tan Zhuo, Wang Jiafeng, Hu Ying, Sun Jiao, Bao Meihua, Huang Ping, Ge Minghua, Chai Young Jun, Zheng Chuanming
Clinical Pharmacy Center, Department of Pharmacy, Zhejiang Provincial People's Hospital; Affiliated People's Hospital, Hangzhou Medical College, Hangzhou, China.
Key Laboratory of Endocrine Gland Diseases of Zhejiang Province, Hangzhou, China.
Gland Surg. 2022 Jan;11(1):245-257. doi: 10.21037/gs-21-908.
Anaplastic thyroid carcinoma (ATC) is an extremely aggressive solid tumor with no effective treatment at present. Because of the rapid growth and aggressiveness, nearly all patients die within six months after developing ATC. Hence, more research regarding novel therapeutic targets for ATC is urgently needed.
Single-cell RNA sequencing data and microarray data of ATC were retrieved from the Gene Expression Omnibus (GEO) database. Cell clustering was performed using the Seurat package. Then, differential expression and functional enrichment analyses were performed. Gene set enrichment analysis (GSEA) was further used to investigate the functional enrichment of lysyl oxidase (LOX) and bone morphogenetic protein-1 (BMP1). The expression levels of LOX and BMP1 were measured using quantitative real-time PCR and Western blot. LOX and BMP1 were knocked down using si-RNAs. Cell proliferation was evaluated by the CCK-8 and clone formation assays. Cell migration and invasion were assessed by the wound healing assay and Transwell assay, respectively.
LOX was upregulated at the single-cell level, as well as in ATC tissues and cell lines. LOX knockdown significantly inhibited ATC cell proliferation. Furthermore, the migration and invasion of ATC cells were remarkably inhibited after LOX inhibition. In addition, BMP1 regulated LOX expression in 8505C cells, while BMP1 overexpression restored the LOX activity blocked by the LOX inhibitor BAPN. BMP1 could also induce the cell proliferation and metastasis of ATC.
LOX/BMP1 mediates the malignant progression of ATC, highlighting the potential application of LOX/BMP1 in the treatment of ATC. This study provides new insights for efficient therapeutic agents based on the LOX/BMP1 axis.
间变性甲状腺癌（ATC）是一种极具侵袭性的实体瘤，目前尚无有效治疗方法。由于其生长迅速且具有侵袭性，几乎所有患者在发生ATC后六个月内死亡。因此，迫切需要对ATC的新型治疗靶点进行更多研究。
从基因表达综合数据库（GEO）中检索ATC的单细胞RNA测序数据和微阵列数据。使用Seurat软件包进行细胞聚类。然后，进行差异表达和功能富集分析。进一步使用基因集富集分析（GSEA）研究赖氨酰氧化酶（LOX）和骨形态发生蛋白-1（BMP1）的功能富集情况。使用定量实时PCR和蛋白质免疫印迹法检测LOX和BMP1的表达水平。使用小干扰RNA（si-RNA）敲低LOX和BMP1。通过CCK-8和克隆形成试验评估细胞增殖。分别通过伤口愈合试验和Transwell试验评估细胞迁移和侵袭能力。
LOX在单细胞水平以及ATC组织和细胞系中均上调。敲低LOX可显著抑制ATC细胞增殖。此外，抑制LOX后，ATC细胞的迁移和侵袭能力明显受到抑制。此外，BMP1在8505C细胞中调节LOX表达，而BMP1过表达可恢复被LOX抑制剂β-氨基丙腈（BAPN）阻断的LOX活性。BMP1还可诱导ATC细胞增殖和转移。
LOX/BMP1介导ATC的恶性进展，突出了LOX/BMP1在ATC治疗中的潜在应用价值。本研究为基于LOX/BMP1轴的高效治疗药物提供了新的见解。
