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[长链非编码RNA SNHG15通过miR-30b-3p的海绵吸附作用调控COX6B1,促进肺腺癌细胞的增殖、迁移和侵袭]

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

作者信息

Gong Xiuying, Hou Shunfu, Zhao Miaomiao, Wang Xiaona, Zhang Zhihan, Liu Qinghua, Yin Chonggao, Li Hongli

机构信息

School of Basic Medical Sciences, Shandong Second Medical University, Weifang 261000, China.

School of Life Sciences and Technology, Shandong Second Medical University, Weifang 261000, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2025 Jul 20;45(7):1498-1505. doi: 10.12122/j.issn.1673-4254.2025.07.16.

Abstract

OBJECTIVES

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 (=0.128, =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.

CONCLUSIONS

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.

摘要

目的

探讨长链非编码RNA SNHG15调控肺腺癌细胞增殖、侵袭和迁移的分子机制。

方法

利用长链非编码RNA微阵列芯片数据集GSE196584和LncBase预测与miR-30b-3p相互作用的长链非编码RNA,并通过在线数据库研究它们与患者预后的关系,之后选择长链非编码核仁RNA宿主基因15(SNHG15)进行进一步分析。采用荧光原位杂交和qRT-PCR检测长链非编码RNA SNHG15的亚细胞定位及其在正常人肺上皮细胞和肺腺癌细胞系中的表达水平。在培养的A549细胞中,用EdU、伤口愈合和Transwell实验评估转染SNHG15敲低质粒(sh-SNHG15)、miR-30b-3p抑制剂或二者共转染后细胞增殖、迁移和侵袭的变化。利用生物信息学分析预测长链非编码RNA SNHG15与COX6B1之间的调控关系,并在转染sh-SNHG15、COX6B1过表达质粒或二者的A549细胞中通过蛋白质免疫印迹和拯救实验验证结果。

结果

长链非编码RNA SNHG15被证明靶向miR-30b-3p,前者在肺腺癌中高表达,且与患者预后不良相关。长链非编码RNA SNHG15定位于细胞质,在A549和NCI-H1299细胞中的表达水平高于BEAS-2B细胞。在A549细胞中,长链非编码RNA SNHG15敲低显著抑制细胞迁移、侵袭和增殖,而miR-30b-3p抑制剂可逆转这些变化。发现长链非编码RNA SNHG15与COX6B1之间存在调控关系,它们的表达水平呈正相关(=0.128,=0.003)。miR-30b-3p敲低明显降低A549细胞中COX6B1的表达,COX6B1过表达使细胞免受长链非编码RNA-SNHG15敲低的抑制作用。

结论

长链非编码RNA SNHG15可能通过竞争性内源RNA机制与COX6B1竞争结合miR-30b-3p,从而影响肺腺癌细胞的增殖、迁移和侵袭。

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