Department of Urology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
J Cell Biochem. 2020 Oct;121(10):4214-4225. doi: 10.1002/jcb.29626. Epub 2020 Jan 3.
The study aimed to investigate the expression and function of bladder cancer (BC) long noncoding RNAs (lncRNAs) using a high-throughput platform. High-throughput sequencing was used to compare the expression profiles of lncRNA in BC and adjacent normal tissues. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), in situ hybridization, gene ontology, and Kyoto Encyclopedia of Genes and Genomes analysis were performed to verify differential expression of lncRNA. The effect of lncRNA overexpression on cellular proliferation, apoptosis, migration, and invasion was analyzed following the transfection of lncRNA overexpressing lentivirus into 5637 and T24 cell lines. The overexpressing cells were subcutaneously injected into nude mice to evaluate their effects on tumor growth. Tumor-associated RNA-binding proteins of lncRNA were analyzed by RNA pull-down combined with mass spectrometry. A total of 93 lncRNA genes were upregulated and 352 lncRNA genes were downregulated in the tissues of patients with BC. Of which, we investigated the function of downregulated lnc-MUC20-9. Overexpression of lnc-MUC20-9 in 5637 and T24 cells resulted in decreased tumor cell viability and cell clones, decreased migration and invasion, and increased apoptosis. Similarly, nude mice bearing lnc-MUC20-9 overexpressing tumor cells exhibited smaller tumor size and volume than that of mice bearing control cells. Mass spectrometry analysis showed that lnc-MUC20-9 binds to ROCK1, an oncogene whose expression was decreased in lnc-MUC20-9 overexpressing cells. The study revealed that lnc-MUC20-9 has the function of inhibiting tumor growth, migration, and invasion. In BC cells, lnc-MUC20-9 binds to ROCK1 and may be involved in lnc-MUC20-9-mediated tumor suppressor function, which might be potential therapeutic targets for BC.
本研究旨在利用高通量平台研究膀胱癌(BC)长链非编码 RNA(lncRNA)的表达和功能。高通量测序用于比较 BC 和相邻正常组织中 lncRNA 的表达谱。采用定量逆转录-聚合酶链反应(qRT-PCR)、原位杂交、基因本体论和京都基因与基因组百科全书分析验证 lncRNA 的差异表达。通过转染 lncRNA 过表达慢病毒,分析 lncRNA 过表达对 5637 和 T24 细胞系细胞增殖、凋亡、迁移和侵袭的影响。将过表达细胞皮下注射到裸鼠中,评估其对肿瘤生长的影响。通过 RNA 下拉结合质谱分析 lncRNA 的肿瘤相关 RNA 结合蛋白。在 BC 患者组织中,93 个 lncRNA 基因上调,352 个 lncRNA 基因下调。其中,我们研究了下调的 lnc-MUC20-9 的功能。lnc-MUC20-9 在 5637 和 T24 细胞中的过表达导致肿瘤细胞活力和细胞克隆减少、迁移和侵袭减少以及凋亡增加。同样,携带 lnc-MUC20-9 过表达肿瘤细胞的裸鼠的肿瘤体积和体积小于携带对照细胞的裸鼠。质谱分析显示,lnc-MUC20-9 与 ROCK1 结合,ROCK1 是一种在 lnc-MUC20-9 过表达细胞中表达降低的癌基因。该研究表明,lnc-MUC20-9 具有抑制肿瘤生长、迁移和侵袭的功能。在 BC 细胞中,lnc-MUC20-9 与 ROCK1 结合,可能参与 lnc-MUC20-9 介导的肿瘤抑制功能,这可能是 BC 的潜在治疗靶点。