Background Cervical cancer is a serious disease with complicated pathogenesis and thus there is an urgent need to find novel targets for the treatment. Recently, long non-coding RNAs (lncRNAs) have emerged as critical factors in tumorigenesis. In this study, we aimed to investigate the mechanism of HAND2 antisense RNA 1 (HAND2-AS1) on the invasion and metastasis of cervical cancer cells. Methods The expression patterns of HAND2-AS1, microRNA-330-5p (miR-330-5p) and leucine zipper down-regulated in cancer 1 (LDOC1) in cervical cancer were characterized by RT-qPCR and western blot analysis. Dual luciferase reporter assay and RIP were applied to verify relationship between HAND2-AS1, miR-330-5p and LDOC1. Fluorescence in situ hybridization (FISH) was used to detect the subcellular localization of HAND2-AS1. Besides, viability, invasion and migration ability of HeLa cells were investigated by cell counting kit-8 (CCK-8) and Transwell assays respectively. Hematoxylin-eosin staining was performed for lymph node metastasis detection. In addition, the tumor growth in nude mice was evaluated. Results Low expression of HAND2-AS1 and LDOC1, and high expression of miR-330-5p were detected in cervical cancer tissues and cells. It was found that binding of HAND2-AS1 to miR-330-5p results in upregulation of LDOC1 expression. Also, overexpressed HAND2-AS1 and LDOC1 or down-regulated miR-330-5p inhibited expression of proliferation-associated proteins Ki-67, PCNA, migration-associated proteins N-cad and invasion-related proteins MMP-2, MMP-9 as well as lymph node metastasis. Moreover, HAND2-AS1 inhibited tumor formation and lymph node metastasis by binding to miR-330-5p in vivo. Conclusion HAND2-AS1 promotes LDOC1 expression by competitively binding to miR-330-5p and consequently inhibiting cervical cancer cell invasion and metastasis. This could facilitate development of therapeutic strategies against cervical cancer.

中文翻译：

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HAND2-AS1 通过 microRNA-330-5p 介导的 LDOC1 抑制宫颈癌细胞的侵袭和转移。

背景 宫颈癌是一种发病机制复杂的严重疾病，迫切需要寻找新的治疗靶点。最近，长链非编码 RNA (lncRNA) 已成为肿瘤发生的关键因素。本研究旨在探讨 HAND2 反义 RNA 1 (HAND2-AS1) 对宫颈癌细胞侵袭和转移的作用机制。方法通过RT-qPCR和蛋白质印迹分析表征宫颈癌中HAND2-AS1、microRNA-330-5p（miR-330-5p）和亮氨酸拉链在癌症1（LDOC1）中下调的表达模式。应用双荧光素酶报告基因测定和 RIP 来验证 HAND2-AS1、miR-330-5p 和 LDOC1 之间的关系。荧光原位杂交 (FISH) 用于检测 HAND2-AS1 的亚细胞定位。此外，生存能力，分别通过细胞计数试剂盒8（CCK-8）和Transwell测定法研究HeLa细胞的侵袭和迁移能力。苏木精-伊红染色用于淋巴结转移检测。此外，评估了裸鼠中的肿瘤生长。结果宫颈癌组织和细胞中检测到HAND2-AS1和LDOC1低表达，miR-330-5p高表达。发现 HAND2-AS1 与 miR-330-5p 的结合导致 LDOC1 表达上调。此外，过表达的 HAND2-AS1 和 LDOC1 或下调的 miR-330-5p 抑制增殖相关蛋白 Ki-67、PCNA、迁移相关蛋白 N-cad 和侵袭相关蛋白 MMP-2、MMP-9 的表达，如以及淋巴结转移。此外，HAND2-AS1 通过在体内与 miR-330-5p 结合来抑制肿瘤形成和淋巴结转移。结论 HAND2-AS1通过竞争性结合miR-330-5p促进LDOC1表达，从而抑制宫颈癌细胞的侵袭和转移。这可以促进针对宫颈癌的治疗策略的发展。