南方医科大学学报

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (7): 1498-1505.doi: 10.12122/j.issn.1673-4254.2025.07.16

• • 上一篇    下一篇

LncRNA SNHG15通过miR-30b-3p调控COX6B1轴促进肺腺癌细胞增殖、迁移和侵袭的分子机制

龚秀莹1(), 侯顺福1, 赵苗苗1, 王晓娜1, 张致涵2, 刘清华1, 尹崇高3, 李洪利1()   

  1. 1.山东第二医科大学，基础医学院，山东 潍坊 261000
    2.山东第二医科大学，生命科学与技术学院，山东 潍坊 261000
    3.山东第二医科大学，护理学院，山东 潍坊 261000
  • 收稿日期:2024-12-17 出版日期:2025-07-20 发布日期:2025-07-17
  • 通讯作者: 李洪利 E-mail:20230105@stu.sdsmu.edu.cn;lihongli@sdsmu.edu.cn
  • 作者简介:龚秀莹，在读硕士研究生，E-mail: 20230105@stu.sdsmu.edu.cn
  • 基金资助:
    国家自然科学基金(82373043);山东省自然科学基金(ZR2022MH311);山东省中医药科技项目(Q-2023012)

LncRNA SNHG15 promotes proliferation, migration and invasion of lung adenocarcinoma cells by regulating COX6B1 through sponge adsorption of miR-30b-3p

Xiuying GONG1(), Shunfu HOU1, Miaomiao ZHAO1, Xiaona WANG1, Zhihan ZHANG2, Qinghua LIU1, Chonggao YIN3, Hongli LI1()   

  1. 1.School of Basic Medical Sciences, Shandong Second Medical University, Weifang 261000, China
    2.School of Life Sciences and Technology, Shandong Second Medical University, Weifang 261000, China
    3.School of Nursing, Shandong Second Medical University, Weifang 261000, China
  • Received:2024-12-17 Online:2025-07-20 Published:2025-07-17
  • Contact: Hongli LI E-mail:20230105@stu.sdsmu.edu.cn;lihongli@sdsmu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(82373043)

RichHTML

20

PDF

115

摘要：

目的 探索LncRNA SNHG15是否可以与COX6B1竞争性结合miR-30b-3p调控肺腺癌细胞增殖、迁移和侵袭。 方法 利用LncRNAs微阵列芯片数据集GSE196584和LncBase预测与miR-30b-3p互用的LncRNA。通过在线数据库查询其与患者预后的关系及其表达量，筛选出长链非编码RNA（LncRNA）核仁RNA宿主基因15（SNHG15）作为接下来的研究对象。FISH检测LncRNA SNHG15的亚细胞定位。使用qRT-PCR检测LncRNA SNHG15在人正常肺上皮细胞和肺腺癌细胞系中的表达水平；将A549细胞分为4组并共转染：sh-NC+miR-NC组（空载质粒+miR-30b-3p对照质粒共转染）、sh-SNHG15+miR-NC组（SNHG15敲低质粒+miR-30b-3p对照质粒共转染）、sh-NC+miR-30b-3p-inhibitor组（空载质粒+miR-30b-3p抑制质粒共转染）、sh-SNHG15+miR-30b-3p-inhibitor组（SNHG15敲低质粒+miR-30b-3p抑制质粒共转染），通过EdU、伤口愈合及Transwell实验检测细胞增殖、迁移与侵袭能力。使用生物信息学数据库预测LncRNA SNHG15与COX6B1的调控关系，Western Blot实验验证敲低LncRNA SNHG15后，COX6B1的表达水平。进一步开展SNHG15与COX6B1的挽救实验，将A549细胞分为4组共转染：sh-NC+CON组（SNHG15对照质粒+COX6B1空载质粒共转染）、sh-SNHG15+oe-CON组（SNHG15敲低质粒+COX6B1空载质粒共转染）、sh-NC+oe-COX6B1组（SNHG15对照质粒+COX6B1过表达质粒共转染）、sh-SNHG15+oe-COX6B1组（SNHG15敲低质粒+COX6B1过表达质粒共转染），进行EdU实验、伤口愈合实验、Transwell实验检测各组A549细胞增殖、迁移、侵袭能力的变化。 结果 LncRNA SNHG15与miR-30b-3p存在靶向关系。LncRNA SNHG15高表达与肺腺癌患者不良预后有关且在肺腺癌组织中高表达。LncRNA SNHG15主要定位在细胞质。LncRNA SNHG15在A549、NCI-H1299细胞中的表达高于在BEAS-2B中的表达（P<0.05）。下调miR-30b-3p可以逆转敲低LncRNA SNHG15导致的A549细胞增殖、迁移和侵袭能力的减弱（P<0.05）。LncRNA SNHG15和COX6B1存在调控关系且呈正相关（r=0.128，P=0.003）。Western blotting实验证实敲低LncRNA SNHG15可以使COX6B1的表达水平下降（P<0.05）。过表达COX6B1可以挽救下调LncRNA SNHG15导致的A549细胞迁移，侵袭和增殖能力的下降（P<0.05）。 结论 LncRNA SNHG15可能通过ceRNA机制与COX6B1竞争性结合miR-30b-3p，从而进一步影响肺腺癌的增殖、迁移、侵袭。

关键词: 肺腺癌, LncRNA SNHG15, miR-30b-3p, COX6B1, 迁移, 侵袭

Abstract:

Objective To explore the molecular mechanism by which lncRNA SNHG15 regulates proliferation, invasion and migration of lung adenocarcinoma cells. Methods The lncRNA microarray chip dataset GSE196584 and LncBase were used to predict the lncRNAs that interact with miR-30b-3p, and their association with patient prognosis were investigated using online databases, after which lncRNA nucleolar RNA host gene 15 (SNHG15) was selected for further analysis. The subcellular localization of lncRNA SNHG15 and its expression levels in normal human lung epithelial cells and lung adenocarcinoma cell lines were detected using fluorescence in situ hybridization and qRT-PCR. In cultured A549 cells, the changes in cell proliferation, migration, and invasion following transfection with a SNHG15 knockdown plasmid (sh-SNHG15), a miR-30b-3p inhibitor, or their co-transfection were assessed with EdU, wound healing, and Transwell assays. Bioinformatics analyses were used to predict the regulatory relationship between lncRNA SNHG15 and COX6B1, and the results were verified using Western blotting and rescue experiments in A549 cells transfected with sh-SNHG15, a COX6B1-overexpressing plasmid, or both. Results LncRNA SNHG15 was shown to target miR-30b-3p, and the former was highly expressed in lung adenocarcinoma, and associated with a poor patient prognosis. LncRNA SNHG15 was localized in the cytoplasm and expressed at higher levels in A549 and NCI-H1299 cells than in BEAS-2B cells. In A549 cells, lncRNA SNHG15 knockdown significantly inhibited cell migration, invasion and proliferation, and these changes were reversed by miR-30b-3p inhibitor. A regulatory relationship was found between lncRNA SNHG15 and COX6B1, and their expression levels were positively correlated (r=0.128, P=0.003). MiR-30b-3p knockdown obviously decreased COX6B1 expression in A549 cells, and COX6B1 overexpression rescued the cells from the inhibitory effects of lncRNA-SNHG15 knockdown. Conclusion LncRNA SNHG15 may compete with COX6B1 to bind miR-30b-3p through a ceRNA mechanism to affect proliferation, migration, and invasion of lung adenocarcinoma cells.

Key words: lung adenocarcinoma, long non-coding RNA SNHG15, miR-30b-3p, COX6B1, migration, invasion