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

doi:10.12122/j.issn.1673-4254.2025.07.16

Abstract

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

Xiuying GONG, Shunfu HOU, Miaomiao ZHAO, Xiaona WANG, Zhihan ZHANG, Qinghua LIU, Chonggao YIN, Hongli LI. LncRNA SNHG15 promotes proliferation, migration and invasion of lung adenocarcinoma cells by regulating COX6B1 through sponge adsorption of miR-30b-3p[J]. Journal of Southern Medical University, 2025, 45(7): 1498-1505.

Figures/Tables 4

Fig.1 Screening for LncRNAs with targeted binding to miR-30b-3p. A: Venn plots showing the intersection of GSE196584 with LncBase database predictions. B-D: Survival curve analysis of LINC00963, THAP9-AS1 and LENG8-AS1. E: Survival curve analysis of MALAT1. F: LncRNA SNHG15 survival curve analysis. G: StarBase database analysis of differential expression of LncRNA SNHG15. H: Database LncLocator prediction of LncRNA SNHG15 cellular sublocilization. I: fluorescence in situ hybridization (FISH) assay for detecting localization of LncRNA SNHG15 in lung adenocarcinoma A549 and NCI-H1299 cells (Scale bar=50 μm). J: TargerScan Website Predicts Binding Sites. K: Dual-luciferase assay to validate the target binding ability of miR-30b-3p to LncRNA SNHG15. **P<0.01 vs miR-NC group.

Fig.2 LncRNA SNHG15 promotes A549 migration, invasion and proliferation ability by targeting miR-30b-3p. A: RT-qPCR to verify LncRNA SNHG15 expression in BEAS-2B, A549 and NCI-H1299 cell lines. B: qRT-PCR for assessing the knockdown efficiency of LncRNA SNHG15. C: EdU experiments for assessing the effect of co-transfection with LncRNA SNHG15 and miR-30b-3p inhibitor on proliferation of lung adenocarcinoma cells (Scale bar=100 μm). D: Wound healing assay for assessing the effect of co-transfection with LncRNA SNHG15 and miR-30b-3p inhibitor on migration of A549 cells (Scale bar=500 μm). E: Transwell assay for assessing the effect of co-transfection with LncRNA SNHG15 and miR-30b-3p inhibitor on invasion and migration ability of lung adenocarcinoma cells (Scale bar=100 μm). **P<0.01, ***P<0.001 vs sh-NC+miR-NC group.

Fig.3 Correlation analysis of LncRNA SNHG15, miR-30b-3p and COX6B1 in LUAD. A: Positive correlation between LncRNA SNHG15 and COX6B1 expressions (r=0.128, P=3.33e-03). B: StarBase database analysis of COX6B1 differential expression. C: Survival curve analysis of COX6B1. D: Western blotting for assessing the effect of knockdown of LncRNA SNHG15 on COX6B1 expression. ***P<0.001 vs NC/A549 group.

Fig.4 LncRNA SNHG15 regulates COX6B1 to promote lung adenocarcinoma cell proliferation, migration, and invasion via sponge adsorption of miR-30b-3p. A: EdU experiments for assessing the effect of co-transfection with COX6B1 and miR-30b-3p on proliferation of lung adenocarcinoma cells (Scale bar=100 μm). B: Wound healing assay for assessing the effect of co-transfection with COX6B1 and miR-30b-3p on migration of A549 cells (Scale bar=500 μm). C: Transwell assay for assessing the effect of co-transfection with COX6B1 and miR-30b -3p on migration and invasion ability of lung adenocarcinoma cells (Scale bar=100 μm). **P<0.01, ***P<0.001, ****P<0.0001 vs sh-NC+CON group.

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