Lin Feng, Yao Xianming, Zhang ShuoShuo, Yang Huajun
Department of Urology, Hangzhou Children's Hospital, Hangzhou City, Zhejiang Province, PR China.
Department of Urology, Heji Hospital of Changzhi Medical College, Changzhi City, Shanxi Province, PR China.
Clinics (Sao Paulo). 2025 May 13;80:100629. doi: 10.1016/j.clinsp.2025.100629. eCollection 2025.
The study explored the molecular mechanism of circNOLC1 influencing Testicular Germ Cell Tumor (TGCT) progression.
The study used TGCT tissue samples and cell lines for investigations. The circNOLC1 and miR-140-5p expression in TGCT tissues were done through RT-qPCR. Analysis of the association of circNOLC1 with TGCT clinic-pathological features was also done. Transfections of circNOLC1, miR-140-5p or Insulin-like GrowthFfactor 1 Receptor (IGF1R)-related sequences or plasmids into TCAM-2 and NCCIT TGCT cells were done. The circNOLC1, miR-140-5p, and IGF1R mRNA expression in cells were done through RT-qPCR. Interaction between miR-140-5p, circNOLC1, and IGF1R was examined using a dual-luciferase reporter assay. MTT assay and colony-forming assay were used to investigate cell proliferation. Apoptosis was determined by flow cytometry. Transwell assay was used to investigate cell invasion. IGF1R protein expression was determined through a western blot.
Increased circNOLC1 in TGCT tissues was correlated with lymph node metastasis, clinical stage, and pathological grade of TGCT patients. CircNOLC1 knock-down inhibited TGCT cell proliferation, colony formation, and invasion, and promotedapoptosis. MiR-140-5p was reduced while IGF1R was upregulated in TGCT tissues and cell lines. Moreover, miR-140-5p mimic could reverse the effect of circNOLC1 knock-down on malignant behaviors of TGCT cells. The authors demonstrated that elevated IGF1R reversed the negative effect of miR-140-5p mimic on TGCT cell proliferation, colony formation, and invasion. CircNOLC1 can act as a sponge of miR-140-5p to up-regulate the IGF1R expression level.
The study highlights that circNOLC1 promotes the progression of TGCT by regulating the miR-140-5p/IGF1R axis.
本研究探讨环状核糖核酸NOLC1（circNOLC1）影响睾丸生殖细胞肿瘤（TGCT）进展的分子机制。
本研究采用TGCT组织样本和细胞系进行研究。通过逆转录定量聚合酶链反应（RT-qPCR）检测TGCT组织中circNOLC1和微小核糖核酸-140-5p（miR-140-5p）的表达。还分析了circNOLC1与TGCT临床病理特征的相关性。将circNOLC1、miR-140-5p或胰岛素样生长因子1受体（IGF1R）相关序列或质粒转染到TCAM-2和NCCIT TGCT细胞中。通过RT-qPCR检测细胞中circNOLC1、miR-140-5p和IGF1R信使核糖核酸（mRNA）的表达。使用双荧光素酶报告基因检测法检测miR-140-5p、circNOLC1和IGF1R之间的相互作用。采用甲基噻唑基四唑（MTT）法和集落形成试验研究细胞增殖。通过流式细胞术测定细胞凋亡。采用Transwell试验研究细胞侵袭。通过蛋白质免疫印迹法测定IGF1R蛋白表达。
TGCT组织中circNOLC1表达增加与TGCT患者的淋巴结转移、临床分期和病理分级相关。敲低circNOLC1可抑制TGCT细胞增殖、集落形成和侵袭，并促进细胞凋亡。在TGCT组织和细胞系中，miR-140-5p表达降低而IGF1R表达上调。此外，miR-140-5p模拟物可逆转敲低circNOLC1对TGCT细胞恶性行为的影响。作者证明，上调IGF1R可逆转miR-140-5p模拟物对TGCT细胞增殖、集落形成和侵袭的负面影响。circNOLC1可作为miR-140-5p的海绵，上调IGF1R表达水平。
本研究强调circNOLC1通过调节miR-140-5p/IGF1R轴促进TGCT进展。
