Department of Oral Histopathology, School and Hospital of Stomatology, China Medical University, Liaoning Province Key Laboratory of Oral Disease, Shenyang, Liaoning, China (mainland).
Central Laboratory Department, School and Hospital of Stomatology, China Medical University, Liaoning Province Key Laboratory of Oral Disease, Shenyang, Liaoning, China (mainland).
Med Sci Monit. 2020 Apr 9;26:e923491. doi: 10.12659/MSM.923491.
BACKGROUND Neural cell adhesion molecule 1 (NCAM1; CD56) and E-cadherin are both involved in cell-cell adhesion and cell development processes, and their dysregulation is associated with various tumors. We hypothesized that dysregulated NCAM1 could suppress the invasive behavior of ameloblastoma (AB), and its expression was regulated by miR-141-3p. MATERIAL AND METHODS Real-time qPCR was performed to examine differences in miR-141-3p expression between AB tissues and normal oral tissues (NOMs). The potential target NCAM1 of miR-141-3p was predicted by bioinformatics analysis, which was validated through dual-luciferase assay. The mRNA and protein levels of NCAM1 were detected by real-time qPCR and Western blot, respectively. Furthermore, the expression and distribution of NCAM1 in AB were investigated through immunohistochemical staining, and immunohistochemical staining of E-cadherin was also performed. After overexpression of NCAM1, the migration of AM-1 cells was examined using wound-healing assay. RESULTS Real-time qPCR results confirmed that miR-141-3p was significantly downregulated in AB tissues. According to bioinformatics analysis, NCAM1 was a target of miR-141-3p, which was confirmed by dual luciferase assay. We found that NCAM1 was significantly upregulated in AB tissues at the mRNA and protein levels. Furthermore, NCAM1 and E-cadherin were mainly expressed on the cell membrane of AB. Downregulation of E-cadherin was found in AB tissues. As shown in wound-healing assay results, NCAM1 overexpression significantly inhibited the invasiveness of AM-1 cells. CONCLUSIONS In this study, highly expressed NCAM1 was found in AB, and it suppressed the migration of AB cells and was regulated by miR-141-3p, suggesting its potential value as a therapeutic target for AB.
神经细胞黏附分子 1（NCAM1；CD56）和 E-钙黏蛋白均参与细胞-细胞黏附和细胞发育过程，其失调与各种肿瘤有关。我们假设失调的 NCAM1 可以抑制成釉细胞瘤（AB）的侵袭行为，其表达受 miR-141-3p 调节。
通过实时 qPCR 检测 miR-141-3p 在 AB 组织和正常口腔组织（NOM）中的表达差异。通过生物信息学分析预测 miR-141-3p 的潜在靶标 NCAM1，并通过双荧光素酶报告基因实验进行验证。通过实时 qPCR 和 Western blot 分别检测 NCAM1 的 mRNA 和蛋白水平。此外，通过免疫组织化学染色研究 NCAM1 在 AB 中的表达和分布，并进行 E-钙黏蛋白的免疫组织化学染色。过表达 NCAM1 后，通过划痕愈合实验检测 AM-1 细胞的迁移。
实时 qPCR 结果证实 miR-141-3p 在 AB 组织中显著下调。根据生物信息学分析，NCAM1 是 miR-141-3p 的靶标，双荧光素酶报告基因实验证实了这一点。我们发现 NCAM1 在 AB 组织中的 mRNA 和蛋白水平均显著上调。此外，NCAM1 和 E-钙黏蛋白主要在 AB 的细胞膜上表达。在 AB 组织中发现 E-钙黏蛋白下调。如划痕愈合实验结果所示，NCAM1 过表达显著抑制 AM-1 细胞的侵袭。
在本研究中，我们发现 AB 中存在高表达的 NCAM1，它抑制 AB 细胞的迁移，并受 miR-141-3p 调节，提示其作为 AB 治疗靶点的潜在价值。
