Zhu Xuqing, Xiang Yanli, Mo Feifei, Jin Lingling
Department of Gastroenterology, Taizhou Municipal Hospital, Zhejiang, China.
Turk J Biol. 2023 Jan 19;47(2):130-140. doi: 10.55730/1300-0152.2648. eCollection 2023.
Compelling evidence has manifested a strong association between aberrant expression of long noncoding RNAs (lncRNAs) and gastric carcinoma (GC) development. Nonetheless, biological impacts of differentially expressed lncRNAs (DElncRNAs) on GC are not scrutinized.
Bioinformatics methods were employed for differential expression analysis and target gene prediction. MTT, colony formation, and Transwell methods were implemented for GC cell proliferation, migration, and invasion assessment. Western blot was implemented to test the protein level. The binding of genes was tested with dual-luciferase and RNA binding protein immunoprecipitation (RIP) approaches.
Noticeably high level of LINC00460 was observed in GC tissues and cells. LINC00460 silencing constrained proliferation, migration, and invasion of GC cells. FISH and nuclear-cytoplasmic separation assays confirmed the main presentation of LINC00460 in the cytoplasm. Bioinformatics predicted that LINC00460 had binding sites to miRNA-143-5p, which was upregulated in GC. Dual luciferase and RIP experiments also confirmed the binding relationship. Concurrent silencing of LINC00460s and miRNA-133-5p rescued the repressive influence of sh-LINC004600 on GC cell proliferation, migration, and invasion. HMGA2 was predicted to be a target gene downstream of miRNA-143-5p, their binding relationship was validated via dual luciferase assays. Silencing HMGA2 constrained GC cell proliferation, invasion, and migration. LINC00460 modulated HMGA2 expression via binding miRNA-143-5p, thereby affecting proliferation, invasion, and migration of GC cells.
These findings validated that LINC00460 could regulate HMGA2 via sponging miRNA-143-5p to facilitate GC proliferation, invasion, and migration, which provides a deeper understanding of lncRNAs in the development of GC.
确凿证据表明长链非编码RNA（lncRNAs）的异常表达与胃癌（GC）发生发展之间存在密切关联。然而，差异表达的lncRNAs（DElncRNAs）对GC的生物学影响尚未得到详细研究。
采用生物信息学方法进行差异表达分析和靶基因预测。运用MTT、集落形成和Transwell方法评估GC细胞的增殖、迁移和侵袭能力。通过蛋白质免疫印迹法检测蛋白水平。采用双荧光素酶和RNA结合蛋白免疫沉淀（RIP）方法检测基因的结合情况。
在GC组织和细胞中观察到LINC00460水平显著升高。LINC00460沉默抑制了GC细胞的增殖、迁移和侵袭。荧光原位杂交（FISH）和核质分离实验证实LINC00460主要存在于细胞质中。生物信息学预测LINC00460与miRNA-143-5p存在结合位点，且miRNA-143-5p在GC中上调。双荧光素酶和RIP实验也证实了这种结合关系。同时沉默LINC00460和miRNA-133-5p可挽救sh-LINC004600对GC细胞增殖、迁移和侵袭的抑制作用。预测HMGA2是miRNA-143-5p下游的靶基因，通过双荧光素酶实验验证了它们的结合关系。沉默HMGA2可抑制GC细胞的增殖、侵袭和迁移。LINC00460通过结合miRNA-143-5p调节HMGA2表达，从而影响GC细胞的增殖、侵袭和迁移。
这些研究结果证实LINC00460可通过吸附miRNA-143-5p来调节HMGA2，促进GC的增殖、侵袭和迁移，这为深入了解lncRNAs在GC发生发展中的作用提供了依据。
