Department of Otolaryngology-Head and Neck Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
J Cell Biochem. 2019 Mar;120(3):3018-3026. doi: 10.1002/jcb.27323. Epub 2018 Dec 12.
Laryngeal cancer (LC) is an increasingly common malignant tumors of head and neck cancer. Aberrant expression of microRNA (miRNA) is closely related with LC development. In the current study, we investigated the biological function and underlying molecular mechanism of miR-384 in LC. The results showed that the miR-384 expression was markedly downregulated in LC tissue and cell lines (TU212 and TU686) as compared with that of adjacent nontumor tissues and a normal human bronchial epithelial cell line. Next, we performed gain-of-function and loss-of-function experiments in the TU212 and TU686 cells by transfecting the cells with miR-384 mimics, miR-384 inhibitor, or miRNA control. Moreover, results showed that miR-384 mimic remarkably inhibited LC cell proliferation, which was notably decreased by miR-384 inhibitor. Furthermore, miR-384 mimics notably increased the amounts of DNA fragmentation from the apoptotic cells (a hallmark of apoptosis) and the caspase-3 activity, whereas miR-384 inhibitor resulted in a decline of DNA fragmentation and the caspase-3 activity compared with its control. In addition, a dual-luciferase reporter assay confirmed that Wnt-induced secreted protein-1 (WISP1) gene was a direct target of miR-384. MiR-384 mimic remarkably inhibited the messenger RNA and protein expression of WISP1, which was upregulated by miR-384 inhibitor as compared to its control. WISP1 knockdown by small interfering RNA inhibited LC cell proliferation and promoted cell apoptosis. WISP1 overexpression partly abrogates the effect of miR-384 overexpression. Taken together, these data indicate that miR-384 regulates LC cell proliferation and apoptosis through targeting WISP1 signaling pathway, providing a novel insight into the LC treatment.
喉癌（LC）是一种日益常见的头颈部癌症恶性肿瘤。miRNA（miR）的异常表达与 LC 的发生发展密切相关。在本研究中，我们研究了 miR-384 在 LC 中的生物学功能及其潜在的分子机制。结果表明，与相邻非肿瘤组织和正常的人支气管上皮细胞系相比，LC 组织和细胞系（TU212 和 TU686）中 miR-384 的表达明显下调。接下来，我们通过转染 miR-384 模拟物、miR-384 抑制剂或 miRNA 对照，在 TU212 和 TU686 细胞中进行了功能获得和功能丧失实验。此外，结果表明 miR-384 模拟物可显著抑制 LC 细胞增殖，而 miR-384 抑制剂则明显降低了 miR-384 抑制剂的增殖。此外，miR-384 模拟物显著增加了来自凋亡细胞（凋亡的标志）的 DNA 片段的量和 caspase-3 的活性，而 miR-384 抑制剂导致与对照相比 DNA 片段和 caspase-3 活性下降。此外，双荧光素酶报告基因实验证实，Wnt 诱导的分泌蛋白-1（WISP1）基因是 miR-384 的直接靶基因。miR-384 模拟物显著抑制 WISP1 的信使 RNA 和蛋白表达，而 miR-384 抑制剂则上调了 WISP1 的表达。用小干扰 RNA 敲低 WISP1 抑制 LC 细胞增殖并促进细胞凋亡。WISP1 过表达部分逆转了 miR-384 过表达的作用。综上所述，这些数据表明，miR-384 通过靶向 WISP1 信号通路调节 LC 细胞增殖和凋亡，为 LC 的治疗提供了新的见解。
