Galang Joelle Noriko, Shen Yefeng, Koitzsch Ulrike, Yu Xiaojie, Eischeid-Scholz Hannah, Bachurski Daniel, Rau Tilman T, Neppl Christina, Herling Marco, Bulimaga Bianca, Vasyutina Elena, Schweiger Michal R, Büttner Reinhard, Odenthal Margarete, Anokhina Maria M
Institute of Pathology, University Hospital of Cologne, 50937 Cologne, Germany.
Center for Molecular Medicine Cologne, University of Cologne, 50937 Cologne, Germany.
Int J Mol Sci. 2023 Sep 12;24(18):13981. doi: 10.3390/ijms241813981.
Lysine-specific demethylase 1 (LSD1) is highly expressed in many cancer types and strongly associated with cancer progression and metastasis. Circular RNAs (circRNAs) are produced by back-splicing and influence the interactive RNA network by microRNA and protein sponging. In the present study, we aimedto identify circRNAs that derive from the LSD1-encoding gene, and to investigate their potential to be released and uptaken by lung cancer versus non-cancer epithelial cells. We identified four circLSD1-RNAs by RT-PCR with divergent primers, followed by sequencing. The expression level of circLSD1-RNAs was then studied by quantitative PCR on cellular and extracellular fractions of lung cancer (PC9) and non-cancer primary small airway epithelial (PSAE) cells. Moreover, we established the transgenic overexpression of circLSD1-RNAs. We show that circLSD1-RNAs are primarily located in the cytoplasm, but are packaged and released from lung cancer and non-cancer cells by extracellular vesicles (EVs) and ribonucleoprotein (RNP) complexes, respectively. Proteomics demonstrated a different protein pattern of EV fractions released from PC9 versus PSAE cells. Importantly, released circLSD1-RNAs were differently taken up by PSAE and PC9 cells. In conclusion, our findings provide primary evidence that circLSD1-RNAs participate in the intercellular communication of lung cancer cells with the tumor environment.
赖氨酸特异性去甲基化酶1（LSD1）在多种癌症类型中高表达，且与癌症进展和转移密切相关。环状RNA（circRNA）通过反向剪接产生，并通过微小RNA和蛋白质海绵作用影响相互作用的RNA网络。在本研究中，我们旨在鉴定源自LSD1编码基因的circRNA，并研究其在肺癌细胞与非癌上皮细胞中的释放和摄取潜力。我们通过使用发散引物的RT-PCR鉴定了四种circLSD1-RNA，随后进行测序。然后通过定量PCR研究circLSD1-RNA在肺癌（PC9）细胞和非癌原代小气道上皮（PSAE）细胞的细胞内和细胞外部分的表达水平。此外，我们建立了circLSD1-RNA的转基因过表达。我们发现circLSD1-RNA主要位于细胞质中，但分别通过细胞外囊泡（EV）和核糖核蛋白（RNP）复合物从肺癌细胞和非癌细胞中包装和释放。蛋白质组学显示，PC9细胞和PSAE细胞释放的EV部分具有不同的蛋白质模式。重要的是，释放的circLSD1-RNA被PSAE细胞和PC9细胞以不同方式摄取。总之，我们的研究结果提供了初步证据，表明circLSD1-RNA参与肺癌细胞与肿瘤环境之间的细胞间通讯。
