Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Nanjing Medical University, Nanjing 211166, Jiangsu, PR China.
Department of Pharmacy, the First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, Anhui, PR China.
Life Sci. 2023 Feb 15;315:121387. doi: 10.1016/j.lfs.2023.121387. Epub 2023 Jan 11.
Exosomes are a subpopulation of extracellular vesicles (EV) derived from multivesicular body (MVB) that transmit various cellular molecular constituents, including long noncoding RNAs (lncRNAs), to promote intercellular communication. Our aim was to investigate the function and mechanism of exosomal LINC00355 in gastric cancer cells.
Exosomal levels of LINC00355 in GC patients and healthy controls were measured by RT-qPCR. The effects of exosomal LINC00355 on GC cell viability, proliferation, migration and invasion were evaluated by CCK8, colony formation, Transwell and wound healing assays. The expression levels of Ki67 in xenograft tumor tissues were confirmed by immunohistochemistry assay, and apoptosis was analyzed by TUNEL apoptosis assay. Western blotting was used to monitor protein expression. RNA immunoprecipitation and RNA pulldown were performed to detect the interaction between LINC00355 and HDAC3. Chromatin immunoprecipitation was used to assess the interaction of HDAC3 with the TP53INP1 promoter.
Exosomal LINC00355 levels were higher in plasma from gastric cancer patients than in plasma from healthy volunteers. Exosomal LINC00355 promoted the proliferation, migration and invasion of gastric cancer cell lines. RNA sequence analysis demonstrated that LINC00355 knockdown downregulated histone deacetylase HDAC3 and upregulated TP53INP1. Mechanistic investigation indicated that exosomal LINC00355 interacted with HDAC3 to suppress TP53INP1 transcription, which promoted epithelial-mesenchymal transition (EMT).
Exosomal LINC00355 plays a pivotal role in regulating EMT to induce the malignant progression of GC. Exosomal LINC00355 could be a promising biomarker in the early diagnosis and prognosis of GC.
外泌体是源自多泡体 (MVB) 的细胞外囊泡 (EV) 的一个亚群，可传递包括长链非编码 RNA (lncRNA) 在内的各种细胞分子成分，从而促进细胞间通讯。我们的目的是研究外泌体 LINC00355 在胃癌细胞中的功能和机制。
通过 RT-qPCR 测量 GC 患者和健康对照者血浆中外泌体 LINC00355 的水平。通过 CCK8、集落形成、Transwell 和划痕愈合实验评估外泌体 LINC00355 对 GC 细胞活力、增殖、迁移和侵袭的影响。免疫组化法验证异种移植肿瘤组织中 Ki67 的表达水平，TUNEL 凋亡法分析细胞凋亡。通过 Western blot 监测蛋白表达。RNA 免疫沉淀和 RNA 下拉实验检测 LINC00355 与 HDAC3 的相互作用。染色质免疫沉淀实验评估 HDAC3 与 TP53INP1 启动子的相互作用。
胃癌患者血浆中外泌体 LINC00355 的水平高于健康志愿者。外泌体 LINC00355 促进了胃癌细胞系的增殖、迁移和侵袭。RNA 序列分析表明，LINC00355 敲低下调了组蛋白去乙酰化酶 HDAC3，上调了 TP53INP1。机制研究表明，外泌体 LINC00355 与 HDAC3 相互作用，抑制 TP53INP1 转录，从而促进上皮-间充质转化 (EMT)。
外泌体 LINC00355 在调节 EMT 以诱导 GC 的恶性进展中发挥关键作用。外泌体 LINC00355 可能成为 GC 早期诊断和预后的有前途的生物标志物。
