Yin Dong, Wei Guo, Sun Xiaoyan, Yin Yan
Department of Dermatology, Shanxi Provincial People's Hospital, Xi'an City, Shaanxi Province, China.
Department of Dermatology, The Second Hospital of Shandong University, Jinan City, Shandong Province, China.
Histol Histopathol. 2022 Oct;37(10):985-997. doi: 10.14670/HH-18-468. Epub 2022 May 11.
Cutaneous squamous cell carcinoma (CSCC) is one of the causes of cancer-related death worldwide. Circular RNAs (circRNAs) play a vital role in the pathological process of many malignant tumors. This study aimed to explore the specific role and potential mechanism of circRNA EBF transcription factor 1 (circEBF1) in CSCC.
The levels of circEBF1, microRNA-1247-5p (miR-1247-5p) and colony stimulating factor 3 (CSF3) were determined by quantitative real-time PCR and Western blot. Cell proliferation was assessed by colony formation assay. Cell migration, invasion and apoptosis were evaluated by Transwell, wound healing, flow cytometry and Western blot assays. Cellular glycolysis, including glucose consumption, lactate production, and ATP level, was detected by commercial kits. The binding relationship between miR-1247-5p and circEBF1 or CSF3 was verified by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay. Xenograft experiment was conducted to analyze tumor growth in vivo.
circEBF1 and CSF3 were up-regulated, while miR-1247-5p was down-regulated in CSCC tissues and cells. Interference of circEBF1 hindered the proliferation, migration, invasion, and glycolysis of CSCC cells and promoted apoptosis. In addition, circEBF1 sponged miR-1247-5p to regulate CSCC cell development. Also, miR-1247-5p suppressed CSCC cell progression by inhibiting CSF3. Moreover, depletion of circEBF1 blocked CSCC tumor growth in vivo.
Down-regulation of circEBF1 impeded CSCC progression by regulating the miR-1247-5p/CSF3 pathway, suggesting that circEBF1 might be a promising therapeutic target for CSCC.
皮肤鳞状细胞癌(CSCC)是全球癌症相关死亡的原因之一。环状RNA(circRNAs)在许多恶性肿瘤的病理过程中起着至关重要的作用。本研究旨在探讨环状RNA EBF转录因子1(circEBF1)在CSCC中的具体作用及潜在机制。
采用定量实时聚合酶链反应和蛋白质免疫印迹法检测circEBF1、微小RNA-1247-5p(miR-1247-5p)和集落刺激因子3(CSF3)的水平。通过集落形成试验评估细胞增殖。采用Transwell、伤口愈合、流式细胞术和蛋白质免疫印迹试验评估细胞迁移、侵袭和凋亡。使用商业试剂盒检测细胞糖酵解,包括葡萄糖消耗、乳酸生成和三磷酸腺苷水平。通过双荧光素酶报告基因试验、RNA免疫沉淀(RIP)试验和RNA下拉试验验证miR-1247-5p与circEBF1或CSF3之间的结合关系。进行异种移植实验以分析体内肿瘤生长情况。
circEBF1和CSF3在CSCC组织和细胞中上调,而miR-1247-5p下调。circEBF1的干扰阻碍了CSCC细胞的增殖、迁移、侵袭和糖酵解,并促进了细胞凋亡。此外,circEBF1通过海绵吸附miR-1247-5p来调节CSCC细胞的发育。而且,miR-1247-5p通过抑制CSF3来抑制CSCC细胞进展。此外,circEBF1的缺失在体内阻断了CSCC肿瘤的生长。
circEBF1的下调通过调节miR-1247-5p/CSF3途径阻碍了CSCC的进展,表明circEBF1可能是CSCC的一个有前景的治疗靶点。