Department of Respiratory and Critical Care Medicine, The First Hospital of Longyan Affiliated to Fujian Medical University, Longyan City, Fujian Province, 364000, China.
Department of Respiratory and Critical Care Medicine, The First Hospital of Longyan Affiliated to Fujian Medical University, Longyan City, Fujian Province, 364000, China.
Mol Cell Probes. 2022 Oct;65:101849. doi: 10.1016/j.mcp.2022.101849. Epub 2022 Aug 18.
As reported, long non-coding RNAs are a pivotal player in lung squamous cell carcinoma (LSCC) progression. We noticed the remarkably upregulated transmembrane-4-l-six-family-19 antisense RNA 1 (TM4SF19-AS1) in LSCC and further demonstrated the function it played in LSCC and the possible molecular mechanism.
Via bioinformatics approach, we evaluated TM4SF19-AS1 and TM4SF19 levels in LSCC tissue, and real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot revealed their mRNA and protein levels in LSCC cells. Cell Counting Kit-8 and colony formation assays analyzed the proliferation ability of LSCC cells, and cell adhesion ability was detected via cell adhesion assay. RNA immunoprecipitation and chromatin immunoprecipitation analyzed the underlying mechanism of TM4SF19-AS1 regulating its target, while methylation-specific PCR indicated the methylation level of TM4SF19-AS1.
TM4SF19-AS1 was markedly upregulated in LSCC. Functional assays revealed that TM4SF19-AS1 could facilitate the proliferation and adhesion of LSCC. Besides, we revealed the mechanism of TM4SF19-AS1 regulation that it directly bound to WD repeat-containing protein 5 (WDR5), and was then recruited to TM4SF19 promoter region, which activated DNA demethylation, thereby suppressing malignant LSCC progression.
Our research demonstrated that TM4SF19-AS1 affected LSCC cell proliferation by recruiting WDR5 to manipulate transmembrane-4-lsix-family-member-19 (TM4SF19), which offers a new observation on LSCC pathogenesis, indicating that TM4SF19-AS1 is able to be a promising target for LSCC treatment.
据报道,长链非编码 RNA 是肺鳞状细胞癌(LSCC)进展的关键因素。我们注意到 LSCC 中跨膜 4 超家族 19 反义 RNA 1(TM4SF19-AS1)显著上调,并进一步证实了其在 LSCC 中的功能及其可能的分子机制。
通过生物信息学方法,我们评估了 LSCC 组织中 TM4SF19-AS1 和 TM4SF19 的水平,并通过实时定量聚合酶链反应(qRT-PCR)和 Western blot 显示了它们在 LSCC 细胞中的 mRNA 和蛋白水平。细胞计数试剂盒-8 和集落形成实验分析了 LSCC 细胞的增殖能力,细胞黏附实验检测了细胞黏附能力。RNA 免疫沉淀和染色质免疫沉淀分析了 TM4SF19-AS1 调节其靶基因的潜在机制,而甲基化特异性 PCR 则显示了 TM4SF19-AS1 的甲基化水平。
TM4SF19-AS1 在 LSCC 中明显上调。功能实验表明,TM4SF19-AS1 可促进 LSCC 的增殖和黏附。此外,我们揭示了 TM4SF19-AS1 调节的机制,即它直接与 WD 重复蛋白 5(WDR5)结合,然后被招募到 TM4SF19 启动子区域,激活 DNA 去甲基化,从而抑制恶性 LSCC 的进展。
我们的研究表明,TM4SF19-AS1 通过招募 WDR5 来影响 LSCC 细胞的增殖,从而操纵跨膜 4 超家族 19 成员 1(TM4SF19),这为 LSCC 发病机制提供了新的观察角度,表明 TM4SF19-AS1 有望成为 LSCC 治疗的新靶点。