Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Oct 10:11:e16242.
doi: 10.7717/peerj.16242. eCollection 2023.

Identification of a novel intermittent hypoxia-related prognostic lncRNA signature and the ceRNA of lncRNA GSEC/miR-873-3p/EGLN3 regulatory axis in lung adenocarcinoma

Peijun Liu #  1 , Long Zhou #  2 , Hao Chen  1 , Yang He  1 , Guangcai Li  2 , Ke Hu  1
Affiliations

Identification of a novel intermittent hypoxia-related prognostic lncRNA signature and the ceRNA of lncRNA GSEC/miR-873-3p/EGLN3 regulatory axis in lung adenocarcinoma

Peijun Liu et al. PeerJ. .

Abstract

Background: Lung adenocarcinoma (LUAD) is still the most prevalent type of respiratory cancer. Intermittent hypoxia can increase the mortality and morbidity associated with lung cancer. Long non-coding RNAs (lncRNAs) are crucial in lung adenocarcinoma. However, the effects of intermittent hypoxia-related long non-coding RNAs (IHRLs) on lung adenocarcinoma are still unknown.

Method: In the current research, eight IHRLs were selected to create a prognostic model. The risk score of the prognostic model was evaluated using multivariate and univariate analyses, and its accuracy and reliability were validated using a nomogram and ROC. Additionally, we investigated the relationships between IHRLs and the immune microenvironment.

Result: Our analysis identified GSEC, AC099850.3, and AL391001.1 as risk lncRNAs, while AC010615.2, AC010654.1, AL513550.1, LINC00996, and LINC01150 were categorized as protective lncRNAs. We observed variances in the expression of seven immune cells and 15 immune-correlated pathways between the two risk groups. Furthermore, our results confirmed the ceRNA network associated with the intermittent hypoxia-related lncRNA GSEC/miR-873-3p/EGLN3 regulatory pathway. GSEC showed pronounced expression in lung adenocarcinoma tissues and specific cell lines, and its inhibition resulted in reduced proliferation and migration in A549 and PC9 cells. Intriguingly, GSEC manifested oncogenic properties by sponging miR-873-3p and demonstrated a tendency to modulate EGLN3 expression favorably.

Conclusion: GSEC acts as an oncogenic lncRNA by interacting with miR-873-3p, modulating EGLN3 expression. This observation underscores the potential of GSEC as a diagnostic and therapeutic target for LUAD.

Keywords: EGLN3; GSEC; Immune microenvironment; Intermittent hypoxia; Lung adenocarcinoma; MiR-873-3p.

PubMed Disclaimer

Conflict of interest statement

The authors declare there are no competing interests.

Figures

Figure 1
Figure 1. The flowchart of the entire research.
Figure 2
Figure 2. Identification of the intermittent hypoxia-related genes (IHRGs) expression and biological functional enrichment research.
(A) Heatmap of the 32 different expression genes (DEG) in LUAD and normal tissues. (B) PPI network of 32 IHRGs. (C) HIF1A, VEGFA, and TP53 are identified as hub genes in the network. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
Figure 3
Figure 3. KEGG and GO analysis of 32 DEGs.
(A–B) GO enrichment of IHRGs in biological processes (BP), cellular component (CC), and molecular function (MF). (C–D) The KEGG analysis revealed the IHRGs.
Figure 4
Figure 4. Identification of intermittent hypoxia-related lncRNAs (IHRLs) and their subsistence analysis.
(A) PCA of all LncRNAs. (B–C) The LASSO Cox algorithm was used to establish a prognosis model. (D) Eight lncRNAs were selected to construct the risk model. (E) Co-expression structure between IHRLs and genes. (E) Sankey diagram of the co-expression network. (F) The PCA visualized the different distribution patterns of patients in eight intermittent hypoxia-related lncRNAs.
Figure 5
Figure 5. Construction of risk model in LUAD patients.
(A–B) The heatmap of the expression of 8 IHRLs. (C–D) The risk score of LUAD patients in the two cohorts. (E–F) The survival status of LUAD patients in the two cohorts. (G–H) The Kaplan–Meier analysis of the OS of LUAD patients.
Figure 6
Figure 6. Prognosis value of the risk model.
(A) Univariate analysis of clinical features. (B) Multivariate analysis of clinical features. (C) The ROC curves of the risk model (risk score: AUC =0.782).
Figure 7
Figure 7. Analysis of immune activity.
(A) Comparison in immune cells of risk groups. (B) GSEA enrichment of oncogenic gene sets. (C-J) The relationship between infiltration of immune cells and the risk model. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
Figure 8
Figure 8. Construction of a ceRNA of lncRNA-miRNA-mRNA.
(A) LncRNA GSEC bound to eight miRNAs. (B) The expression of lncRNA GSEC. (C) Kaplan–Meier survival of lncRNA GSEC. (D)Venn diagram identified the downstream targets in the TargetScan database and IHRLs. (E)The expression of EGLN3 in the GEPIA database. (F) The ceRNA of lncRNA GSEC/miR-873-3p/EGLN3 regulatory axis. ∗p < 0.05; ∗∗∗p < 0.001.
Figure 9
Figure 9. The expression and the effect of lncRNA GSEC, miR-873-3p, and EGLN3 in LUAD.
(A) Expression of GSEC in different LUAD cell lines compared to normal astrocytes. (B) Expression of miR-873-3p in different LUAD cell lines compared to normal astrocytes. (C) Expression of EGLN3 in different LUAD cell lines compared to normal astrocytes. (D) Expression of EGLN3 at the western blot level. (E) The efficiency of GSEC knockdown (si-GSEC). (F-G) The proliferation of cells by CCK-8 assays. (H-I) The migration of cells by wound healing assays. (J-K) The cloning capacity of cells. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
Figure 10
Figure 10. LncRNA GSEC acted as ceRNA for miR-873-3p.
(A) Expression of miR-873-3p following si-GSEC transfection. (B) Expression of GSEC following overexpression of miR-873-3p. (C) The predicted interacting sites. (D–E) The relationship between GSEC and miR-873-3p was performed by Dual-Luciferase reporter assay. (F–G) The immunoprecipitation of GSEC and miR-873-3p was examined by the RIP experiment. ∗∗p < 0.01; ∗∗∗p < 0.001.
Figure 11
Figure 11. MiR-873-3p reversed the lncRNA GSEC knockdown effects on LUAD cells.
(A) The expression of miR-873-3p. (B–C) The proliferation of cells detected by CCK-8 assays. (D–E) The migration of cells detected by wound healing assays. (F–G) The cloning capacity of cells was identified by colony formation assay. ∗∗p < 0.01; ∗∗∗p < 0.001; vs. ctrl inhibitor; #p < 0.05; ##p < 0.01 vs. si-GSEC+ctrl inhibitor.
Figure 12
Figure 12. EGLN3 was a downstream target of miR-873-3p and lncRNA GSEC sponged miR-873-3p to upregulate EGLN3.
(A) The predicted interacting sites. (B–C) The relationship between EGLN3 and miR-873-3p was performed by Dual-Luciferase reporter assay.∗∗p < 0.01; vs. ctrl mimics. (D–F) Expression of EGLN3 following miR-873-3p overexpression. ∗∗p < 0.01; ∗∗∗p < 0.001; vs. ctrl mimics. (G–I) Expression of EGLN3 following si-GSEC or si-GSEC+miR-873-3p inhibitor. ∗∗∗p < 0.001; vs. ctrl inhibitor; #p < 0.05; ##p < 0.01 vs. si-GSEC+ctrl inhibitor.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Alinejad V, Dolati S, Motallebnezhad M, Yousefi M. The role of IL17B-IL17RB signaling pathway in breast cancer. Biomedicine & Pharmacotherapy. 2017;88:795–803. doi: 10.1016/j.biopha.2017.01.120. - DOI - PubMed
    1. Almendros I, Gozal D. Intermittent hypoxia and cancer: undesirable bed partners? Respiratory Physiology & Neurobiology. 2018;256:79–86. doi: 10.1016/j.resp.2017.08.008. - DOI - PubMed
    1. Chen A, Sceneay J, Gödde N, Kinwel T, Ham S, Thompson EW, Humbert PO, A Möller. Intermittent hypoxia induces a metastatic phenotype in breast cancer. Oncogene. 2018;37(31):4214–4225. doi: 10.1038/s41388-018-0259-3. - DOI - PubMed
    1. Chen J, Zhu H, Chen Q, Yang Y, Chen M, Huang J, Chen M, Lian N. The role of ferroptosis in chronic intermittent hypoxia-induced lung injury. BMC Pulmonary Medicine. 2022;22(1):488. doi: 10.1186/s12890-022-02262-x. - DOI - PMC - PubMed
    1. Chen L, Qiu CH, Chen Y, Wang Y, Zhao JJ, Zhang M. LncRNA SNHG16 drives proliferation, migration, and invasion of lung cancer cell through modulation of miR-520/VEGF axis. European Review for Medical and Pharmacological Sciences. 2020;24(18):9522–9531. doi: 10.26355/eurrev_202009_23037. - DOI - PubMed

Publication types

LinkOut - more resources