Deng Jie, Wang Yiqun, Zhang Sheng, Chen Lin
Department of Stomatology, the Second Xiangya Hospital, Central South University, Changsha, China.
Department of Stomatology, the Second Xiangya Hospital, Central South University, Changsha, China.
Arch Oral Biol. 2023 Nov;155:105790. doi: 10.1016/j.archoralbio.2023.105790. Epub 2023 Aug 16.
This study aims to investigate the role of the long non-coding RNA-AC103563.8 (lncRNA) in promoting oral squamous cell carcinoma (OSCC) development and to conduct preliminary research on its mechanism.
Microarray technology were used to screen out a lncRNA significantly upregulated in OSCC. Fluorescence in situ hybridization was used to analyze the position of lncRNA-AC103563.8 in cells. A Cal-27 cell line with knockout of the lncRNA-AC103563.8 gene was constructed. Transwell assay and tumor xenograft experiment was used to determine the metastasis and invasion of the cell. Detection of mutations in genes encoding myelin and lymphocyte proteins (MAL) by pyrosequencing. Identification of RNA-Binding Proteins by Mass Spectrometry (ChIRP-MS) experiments were carried out to enrich the proteins that directly bind to lncRNA-AC103563.8. Bioinformatics was used to analyze the target proteins. Some of the selected proteins were verified by parallel reaction monitoring (PRM) to confirm their binding to lncRNA-AC103563.8.
lncRNA-AC103563.8 is upregulated in OSCC tissue and the presence of lncRNA-AC103563.8 in both the nucleus and the cytoplasm. lncRNA-AC103563.8 promoted OSCC cell invasion and metastasis. Methylation occurs in MAL gene promoter. ChIRP-MS identified 330 proteins binding to lncRNA-AC103563.8, and bioinformatics analysis showed that they were involved in a variety of biological processes. PRM experiments confirmed some protein directly bound to lncRNA-AC103563.8.
lncRNA-AC103563.8 is a functional lncRNA that promotes OSCC development by acting on MAL or interacting with other tumor-related proteins. This study also indicates that this lncRNA may exert regulatory functions in OSCC and is a potential target for OSCC therapy.
本研究旨在探讨长链非编码RNA-AC103563.8(lncRNA)在促进口腔鳞状细胞癌(OSCC)发展中的作用,并对其机制进行初步研究。
采用微阵列技术筛选出在OSCC中显著上调的lncRNA。利用荧光原位杂交分析lncRNA-AC103563.8在细胞中的位置。构建lncRNA-AC103563.8基因敲除的Cal-27细胞系。采用Transwell实验和肿瘤异种移植实验测定细胞的转移和侵袭能力。通过焦磷酸测序检测髓磷脂和淋巴细胞蛋白(MAL)编码基因的突变。进行质谱(ChIRP-MS)实验鉴定RNA结合蛋白,以富集直接与lncRNA-AC103563.8结合的蛋白质。利用生物信息学分析靶蛋白。通过平行反应监测(PRM)验证部分选定蛋白与lncRNA-AC103563.8的结合。
lncRNA-AC103563.8在OSCC组织中上调,且在细胞核和细胞质中均有存在。lncRNA-AC103563.8促进OSCC细胞侵袭和转移。MAL基因启动子发生甲基化。ChIRP-MS鉴定出330种与lncRNA-AC103563.8结合的蛋白质,生物信息学分析表明它们参与多种生物学过程。PRM实验证实部分蛋白直接与lncRNA-AC103563.8结合。
lncRNA-AC103563.8是一种功能性lncRNA,通过作用于MAL或与其他肿瘤相关蛋白相互作用促进OSCC发展。本研究还表明该lncRNA可能在OSCC中发挥调节功能,是OSCC治疗的潜在靶点。