ABSTRACT

Long non-coding RNA (lncRNA) X inactive specific transcript (XIST) has been identified as an oncogenic lncRNA in a series of human cancers, including esophageal squamous cell carcinoma (ESCC). In this study, we aimed to further explore the underlying mechanism of XIST on ESCC progression. qRT-PCR assay was used to determine the levels of XIST and miR-129-5p. Western blot analysis was performed to assess cyclin D1 (CCND1) expression. Bioinformatic analysis was performed using starBase v2.0 software. Dual-luciferase reporter and RNA immunoprecipitation assays were employed to confirm the interaction between XIST and miR-129-5p or miR-129-5p and CCND1. Cell cycle progression and apoptosis were measured by flow cytometric analysis, and cell migration and invasion were detected by transwell assay. Mouse studies were used to observe the effect of XIST silencing on tumor growth in vivo. Our results indicated that XIST was upregulated and miR-129-5p was downregulated in ESCC. XIST silencing or miR-129-5p overexpression repressed cell cycle progression, proliferation, migration, invasion, and promoted the apoptosis in ESCC cells. Moreover, XIST directly interacted with miR-129-5p and repressed miR-129-5p expression. MiR-129-5p mediated the regulatory effect of XIST on ESCC cell progression in vitro, and XIST promoted CCND1 expression by sponging miR-129-5p. Additionally, XIST silencing inhibited tumor growth in vivo. Our findings suggested that XIST silencing repressed the progression of ESCC at least partly through regulating the miR-129-5p/CCND1 axis. Targeting XIST might be a potential therapeutic strategy for ESCC treatment.

摘要

长链非编码 RNA (lncRNA) X 非活性特异性转录本 (XIST) 已被鉴定为一系列人类癌症中的致癌 lncRNA，包括食管鳞状细胞癌 (ESCC)。在本研究中，我们旨在进一步探索 XIST 对 ESCC 进展的潜在机制。qRT-PCR 测定用于确定 XIST 和 miR-129-5p 的水平。进行蛋白质印迹分析以评估细胞周期蛋白 D1 (CCND1) 表达。使用starBase v2.0软件进行生物信息学分析。采用双荧光素酶报告基因和 RNA 免疫沉淀分析来确认 XIST 与 miR-129-5p 或 miR-129-5p 与 CCND1 之间的相互作用。通过流式细胞术分析测量细胞周期进程和细胞凋亡，并通过Transwell测定检测细胞迁移和侵袭。体内。我们的结果表明 XIST 在 ESCC 中上调而 miR-129-5p 下调。XIST 沉默或 miR-129-5p 过表达抑制细胞周期进程、增殖、迁移、侵袭，并促进 ESCC 细胞的凋亡。此外，XIST 直接与 miR-129-5p 相互作用并抑制 miR-129-5p 表达。MiR-129-5p在体外介导 XIST 对 ESCC 细胞进展的调节作用，XIST通过海绵 miR-129-5p 促进 CCND1 表达。此外，XIST 沉默抑制了体内肿瘤的生长。我们的研究结果表明，XIST 沉默至少部分通过调节 miR-129-5p/CCND1 轴来抑制 ESCC 的进展。靶向 XIST 可能是 ESCC 治疗的潜在治疗策略。