黄晔 赖蔚菁 黄如 刘佳华

[摘要] 目的 研究lncRNA OCPAT1调控miR-320d/YOD1信号通路在卵巢癌发生发展中的作用及机制。 方法 2019年1月至2020年12月应用TCGA和GTEX数据库筛选，通过qRT-PCR检测确认lncRNA AC007405.3（OCPAT1）在卵巢癌组织和细胞中的表达;在ES-2细胞中敲减OCPAT1，通过实验验证，OCPAT1与miR-320d/YOD1的靶向调控关系。 结果 通过GTEX和TCGA数据库及qRT-PCR检测，确认OCPAT1高表达;敲低OCPAT1后，MTT增殖显示敲减后细胞增殖减低，EdU细胞增殖显示，敲减后处于复制期的细胞减少，流式细胞周期显示敲减后细胞停滞在G0/G1期，流式周期显示敲减后细胞凋亡增加，Transwell migration实验显示，敲减后抑制卵巢癌细胞ES-2的迁移能力，Matrigel invasion 实验显示敲减后抑制卵巢癌细胞ES-2的侵袭能力。双荧光素酶报告基因实验证实，OCPAT1可以靶向作用于miR-320d。 结论 lncRNA OCPAT1可能通过靶向调控miR-320d/YOD1信号通路调控卵巢癌细胞增殖、迁移及侵袭，进而促进卵巢癌的发生及发展。

[关键词] LncRNA OCPAT1;miR-320d/YOD1信号通路;卵巢癌;增殖;迁移;侵袭

[中图分类号] R737.31          [文献标识码] A          [文章编号] 1673-9701（2021）33-0041-04

[Abstract] Objective To study the role and mechanism of lncRNA OCPAT1 in regulating the miR-320d/YOD1 signaling pathway in the development and progression of ovarian cancer. Methods From January 2019 to December 2020， the expression of lncRNA AC007405.3 （OCPAT1） in ovarian cancer tissues and cells was confirmed by RT-qPCR using TCGA and GTEX databases screening. OCPAT1 was knocked out in ES-2 cells， and the targeted regulatory relationship between OCPAT1 and miR-320d/YOD1 was verified by experiments. Results High expression of OCPAT1 was confirmed by GTEX and TCGA databases and RT-qPCR detection. After OCPAT1 knockdown， MTT proliferation showed decreased cell proliferation after knockdown. the EdU cell proliferation showed decreased cells in the replication phase after knockdown. flow cytometry cycle showed that cells stagnated in the G0/G1 phase after knockdown; flow cytometry cycle showed increased cell apoptosis after knockdown. The Transwell migration experiment showed that knockout inhibited the migration ability of ovarian cancer cell ES-2， and the Matrigel invasion experiment showed that knockout inhibited the invasion ability of ovarian cancer cell ES-2. Dual luciferase reporter assay confirmed that OCPAT1 could target miR-320d. Conclusion LncRNA OCPAT1 may regulate the proliferation， metastasis and invasion of ovarian cancer cells through the targeted regulation of the mir-320s/YOD1 signaling pathway， thus promoting the occurrence and development of the ovarian cancer.

[Key words] LncRNA OCPAT1; miR-320d/YOD1 signaling pathway; Ovarian cancer; Proliferation; Metastasis; Invasion

卵巢癌是婦科三大恶性肿瘤之一[1]，发病率仅次于子宫颈癌和子宫内膜癌，居第3位，死亡率却居女性生殖细胞肿瘤的首位[2]。临床上急需卵巢癌早期的肿瘤标志物用于筛查，有利于为卵巢癌的治疗提供理论依据[3]。长链非编码RNA（lncRNA）对卵巢癌的发生和发展起重要的作用，有望为卵巢癌的治疗提供新思路[4]。研究表明，lncRNA AC007405.3在肿瘤细胞高表达，lncRNA AC007405.3可成为卵巢癌诊断的指标，及早期筛查。本研究将该lncRNA命名为卵巢癌进展相关转录本1（OCPAT1）。荧光原位杂交技术（FISH）实验显示，OCPAT1主要定位在细胞质，目前挑选miR-320d作为OCPAT1潜在的靶向微小RNA（miRNA）， 同时miR-320d靶向YOD1。……