The emerging role of microRNA-935 (miR-935) in modulating cancer progression has been recognized. However, its role in regulating choriocarcinoma (CCA) development and progression remains unknown. The present work aims to reveal the effect of miR-935 on CCA cell tumor properties and the related mechanism. The RNA expression of methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit (METTL3), miR-935, and gap junction protein alpha 1 (GJA1) was detected by quantitative real-time polymerase chain reaction. Protein expression of GJA1, Ki67, and METTL3 was measured by western blotting and immunohistochemistry assays. CCK-8 and colony formation were used to analyze cell proliferation. Transwell assays were performed to assess cell migration and invasion. Angiogenesis was investigated by tube formation assay. Xenograft mouse model assay was used to determine miR-935-mediated effect on tumor formation in vivo. The luciferase reporter assay and RNA pull-down assay were used to verify the relationship between miR-935 and GJA1. MeRIP assay was used to analyze the m6A methylation of pri-miR-935. MiR-935 expression was significantly upregulated in CCA tissues and cells when compared with control groups. MiR-935 overexpression promoted CCA cell proliferation, migration, invasion, and tube formation and tumor tumorigenesis in vitro and in vivo, but miR-935 knockdown showed the opposite effects. In addition, miR-935 targeted GJA1 and mediated CCA cell tumor properties by negatively regulating GJA1 expression. METTL3 promoted miR-935 maturation by inducing m6A methylation of pri-miR-935, and its overexpression contributed to CCA cell tumor properties through the regulation of miR-935. METTL3 promoted choriocarcinoma progression by m6A-dependently activating the miR-935/GJA1 pathway.

中文翻译：

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METTL3 通过以 m6A 依赖性方式激活 miR-935/GJA1 通路促进绒毛膜癌进展

microRNA-935 (miR-935) 在调节癌症进展中的新兴作用已得到认可。然而，其在调节绒毛膜癌（CCA）发生和进展中的作用仍然未知。本工作旨在揭示miR-935对CCA细胞肿瘤特性的影响及相关机制。通过实时定量聚合酶链式反应检测甲基转移酶3、N6-腺苷-甲基转移酶复合体催化亚基（METTL3）、miR-935和间隙连接蛋白α1（GJA1）的RNA表达。通过蛋白质印迹和免疫组织化学测定来测量 GJA1、Ki67 和 METTL3 的蛋白表达。CCK-8和集落形成用于分析细胞增殖。进行 Transwell 测定以评估细胞迁移和侵袭。通过管形成测定研究血管生成。异种移植小鼠模型测定用于确定 miR-935 介导的对体内肿瘤形成的影响。使用荧光素酶报告基因测定和RNA Pull-down测定来验证miR-935和GJA1之间的关系。MeRIP 测定用于分析 pri-miR-935 的 m6A 甲基化。与对照组相比，CCA 组织和细胞中 MiR-935 的表达显着上调。miR-935过表达在体外和体内促进CCA细胞增殖、迁移、侵袭、管形成和肿瘤发生，但miR-935敲低则表现出相反的作用。此外，miR-935靶向GJA1并通过负向调节GJA1表达来介导CCA细胞肿瘤特性。METTL3 通过诱导 pri-miR-935 的 m6A 甲基化来促进 miR-935 成熟，其过表达通过调节 miR-935 促进 CCA 细胞肿瘤特性。METTL3 通过 m6A 依赖性激活 miR-935/GJA1 通路促进绒毛膜癌进展。