Environment-Omics-Disease Research Center, China Medical University Hospital, China Medical University, No. 2 Yude Road, Taichung, 40447, Taiwan.
Graduate Institute of Biomedical Sciences, China Medical University, Taichung, 40402, Taiwan.
J Mol Med (Berl). 2021 Sep;99(9):1323-1334. doi: 10.1007/s00109-021-02095-x. Epub 2021 Jun 7.
Studies suggested that long noncoding HAR1A RNA may be a tumor suppressor, but its association with oral cancer remains unclear. Here, we show the functional role and mechanisms of HAR1A in oral cancer progression. Microarray analysis was performed to screen the related candidates of long noncoding RNA (lncRNA) in human monocytes. Following lncRNA HAR1A, the regulation of HAR1A, ALPK1, myosin IIA, and BRD7 was tested using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) in oral cancer cells. The inflammatory and epithelial-to-mesenchymal transition marker expressions were analyzed using enzyme-linked immunosorbent assay and western blot. Phenotypic experiments were verified by colony formation assay, transwell migration assay, and Annexin V-apoptotic assay. In the nuclei of cancer cells, HAR1A functions upstream of signaling pathways and knockdown of HAR1A promoted ALPK1 expression and downregulated BRD7 resulting in inflammation and oral cancer progression. In monocytes, the expressions of TNF-α and CCL2 were increased following HAR1A knockdown and reduced following ALPK1 knockdown. HAR1A knockdown upregulated the expression of ALPK1, slug, vimentin, fibronectin, and N-cadherin but reduced the expression of E-cadherin in oral cancer cells. Myosin IIA was primarily located in the cytoplasm and that its decrease in the nuclei of oral cancer cells was likely to demonstrate suppressive ability in late-stage cancer. Our findings suggest that the HAR1A, BRD7, and myosin IIA are tumor suppressors while ALPK1 has oncogene-like property in the nucleus and is involved in inflammation and oral cancer progression. More research for HAR1A activators or ALPK1 inhibitors is required to develop potential therapeutic agents for advanced oral cancer. KEY MESSAGES: lncRNA HAR1A, BRD7, and myosin IIA are tumor suppressors whereas ALPK1 has an oncogenic-like property in the nucleus. lncRNA HAR1A/ALPK1/BRD7/myosin IIA axis plays a critical role in the progression of oral cancer. lncRNA HAR1A localizes upstream of signaling pathways to inhibit ALPK1 expression and then upregulated BRD7. lncRNA HAR1A and ALPK1 are involved in cancer progression via epithelial-to-mesenchymal transition regulations. ALPK1 inhibitors are potential kinase-targeted therapeutic agents for patients with advanced oral cancer.
研究表明，长链非编码 HAR1A RNA 可能是一种肿瘤抑制因子，但它与口腔癌的关系尚不清楚。在这里，我们展示了 HAR1A 在口腔癌进展中的功能作用和机制。通过人单核细胞的微阵列分析筛选长链非编码 RNA（lncRNA）的相关候选物。在口腔癌细胞中，通过逆转录定量聚合酶链反应（RT-qPCR）测试 HAR1A、ALPK1、肌球蛋白 IIA 和 BRD7 的调控。通过酶联免疫吸附试验和 Western blot 分析炎症和上皮-间充质转化标志物的表达。通过集落形成试验、Transwell 迁移试验和 Annexin V-凋亡试验验证表型实验。在癌细胞的核内，HAR1A 在上游信号通路中发挥作用，敲低 HAR1A 可促进 ALPK1 的表达，下调 BRD7，从而导致炎症和口腔癌的进展。在单核细胞中，HAR1A 敲低后 TNF-α 和 CCL2 的表达增加，而 ALPK1 敲低后表达减少。HAR1A 敲低可上调口腔癌细胞中 ALPK1、slug、波形蛋白、纤连蛋白和 N-钙黏蛋白的表达，降低 E-钙黏蛋白的表达。肌球蛋白 IIA 主要位于细胞质中，口腔癌细胞核内肌球蛋白 IIA 的减少可能表明其在晚期癌症中具有抑制能力。我们的研究结果表明，HAR1A、BRD7 和肌球蛋白 IIA 是肿瘤抑制因子，而 ALPK1 在核内具有癌基因样特性，参与炎症和口腔癌的进展。需要进一步研究 HAR1A 激活剂或 ALPK1 抑制剂，以开发用于晚期口腔癌的潜在治疗药物。关键信息：lncRNA HAR1A、BRD7 和肌球蛋白 IIA 是肿瘤抑制因子，而核内的 ALPK1 具有癌基因样特性。lncRNA HAR1A/ALPK1/BRD7/myosin IIA 轴在口腔癌的进展中起着关键作用。lncRNA HAR1A 定位于信号通路的上游，抑制 ALPK1 的表达，然后上调 BRD7。lncRNA HAR1A 和 ALPK1 通过上皮-间充质转化调节参与癌症进展。ALPK1 抑制剂是晚期口腔癌患者潜在的激酶靶向治疗药物。
