背景:越来越多的证据表明,长链非编码RNA(lncRNA)在癌症发展中起着重要作用。然而,大多数lncRNAs在人类胃癌中的功能仍未完全了解。这里,我们探索了一种新的c-Myc激活的lncRNA的作用,LINC01050,在胃癌进展中。
方法:使用癌症基因组图谱数据集评估LINC01050在胃癌中的表达。通过功能增益和功能丧失实验结合细胞计数试剂盒-8测定研究了其在胃癌中的功能,菌落形成试验,Transwell分析,流式细胞术,蛋白质印迹分析,和异种移植肿瘤和小鼠转移模型。通过生物信息学筛选潜在的LINC01050转录激活剂,并通过染色质免疫沉淀和荧光素酶测定进行验证。通过生物信息学分析预测LINC01050和miR-7161-3p与miR-7161-3p的靶标之间的相互作用,并通过荧光素酶测定法证实。RNA免疫沉淀,RNA下拉,和救援实验。
结果:LINC01050在胃癌中显著上调,高表达与不良预后呈正相关。发现转录因子c-Myc直接与LINC01050启动子区结合并激活其转录。此外,证实LINC01050的过表达可促进胃癌细胞增殖,迁移,入侵,以及体外上皮-间质转化和体内肿瘤生长。同时,其敲除抑制胃癌细胞增殖,迁移,入侵,以及体外上皮-间质转化以及体内肿瘤的生长和转移。此外,机制研究表明,LINC01050作为分子海绵吸收细胞溶质miR-7161-3p,这降低了miR-7161-3p介导的SPZ1的翻译抑制,从而有助于胃癌的进展。
结论:综合来看,我们的结果鉴定了一种新的胃癌相关lncRNA,LINC01050,由c-Myc激活。LINC01050可能被认为是胃癌的潜在治疗靶点。
BACKGROUND: Growing evidence shows that long non-coding RNAs (lncRNAs) play significant roles in cancer development. However, the functions of most lncRNAs in human gastric cancer are still not fully understood. Here, we explored the role of a novel c-Myc-activated lncRNA, LINC01050, in gastric cancer progression.
METHODS: The expression of LINC01050 in the context of gastric cancer was assessed using The Cancer Genome Atlas datasets. Its functions in gastric cancer were investigated through gain- and loss-of-function experiments combined with the Cell Counting Kit-8 assays, colony-forming assays, Transwell assays, flow cytometry, Western blot analyses, and xenograft tumor and mouse metastasis models. Potential LINC01050 transcription activators were screened via bioinformatics and validated by chromatin immunoprecipitation and luciferase assays. The interaction between LINC01050 and miR-7161-3p and the targets of miR-7161-3p were predicted by bioinformatics analysis and confirmed by a luciferase assay, RNA immunoprecipitation, RNA pull-down, and rescue experiments.
RESULTS: LINC01050 was significantly up-regulated in gastric cancer, and its high expression was positively correlated with a poor prognosis. The transcription factor c-Myc was found to directly bind to the LINC01050 promoter region and activate its transcription. Furthermore, overexpression of LINC01050 was confirmed to promote gastric cancer cell proliferation, migration, invasion, and epithelial-mesenchymal transition in vitro and tumor growth in vivo. At the same time, its knockdown inhibited gastric cancer cell proliferation, migration, invasion, and epithelial-mesenchymal transition in vitro along with tumor growth and metastasis in vivo. Moreover, mechanistic investigations revealed that LINC01050 functions as a molecular sponge to absorb cytosolic miR-7161-3p, which reduces the miR-7161-3p-mediated translational repression of
SPZ1, thus contributing to gastric cancer progression.
CONCLUSIONS: Taken together, our results identified a novel gastric cancer-associated lncRNA, LINC01050, which is activated by c-Myc. LINC01050 may be considered a potential therapeutic target for gastric cancer.