Supplementary Material for: Long Non-Coding RNA Expression Profiles for the Characterization of Different Bladder Cancer Grade

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Background/Aims: Bladder cancer (BC) is one of the most frequent urologic tumors worldwide. However, long non-coding RNA(lncRNA) expression profiles in BC progression remain unclear. This study aimed to explore lncRNA expression profiles in different grades of bladder cancer and normal urothelium tissues. Methods: We performed high-throughput sequencing in BC tissues of different grade and obtained the expression profiles of its lncRNAs. Then, aberrantly expressed lncRNAs were validated by quantitative reverse transcription polymerase chain reaction (RT-PCR). Gene Ontology (GO) and pathway analyses were used to investigate the potential function of these lncRNAs. Co-expresson network was constructed to explore the relationship between lncRNAs and target mRNAs. Results: We identified 252 aberrantly expressed lncRNAs in high-grade BC while compared to low-grade BC, and 269 lncRNAs in high-grade BC while compared to normal urothelium. Notably, we found 33 overlapped lncRNAs. Subsequently, 7 lncRNAs were selected from the overlapped part and confirmed by RT-PCR. GO and pathway analyses showed that these dysregulated lncRNAs participated in cell migration, cell adhesion, as well as Ras signaling pathway. Co-expression network and The Cancer Genome Atlas (TCGA) data showed LUCAT1 and CCNB1 had positive relationship in regulating the progress of bladder cancer. Conclusion: Our findings revealed the significant role of lncRNAs in the development process of bladder cancer.

背景与目的：膀胱癌（BC）是全球范围内最为常见的泌尿肿瘤之一。然而，膀胱癌进展过程中的长链非编码RNA（long non-coding RNA, lncRNA）表达谱仍未明确。本研究旨在探讨不同分级膀胱癌与正常尿路上皮组织中的lncRNA表达谱。方法：我们对不同分级的膀胱癌组织进行高通量测序，获取其lncRNA的表达谱。随后通过定量逆转录聚合酶链反应（quantitative reverse transcription polymerase chain reaction, RT-PCR）验证差异表达的lncRNA。采用基因本体论（Gene Ontology, GO）及通路分析，探究上述lncRNA的潜在功能，并构建共表达网络以解析lncRNA与靶mRNA之间的调控关系。结果：相较于低分级膀胱癌，我们在高分级膀胱癌中鉴定出252个差异表达的lncRNA；相较于正常尿路上皮组织，高分级膀胱癌中存在269个差异表达的lncRNA。值得注意的是，本研究发现其中存在33个重叠的lncRNA。随后从重叠的lncRNA中选取7个，并通过RT-PCR完成验证。GO与通路分析结果显示，这些异常表达的lncRNA参与细胞迁移、细胞黏附以及Ras信号通路过程。共表达网络与癌症基因组图谱（The Cancer Genome Atlas, TCGA）数据表明，LUCAT1与CCNB1在调控膀胱癌进展过程中呈正相关关系。结论：本研究结果揭示了lncRNA在膀胱癌发生发展进程中的关键作用。