doi: 10.1111/1759-7714.13374.
Epub 2020 Mar 9.
LncRNA SNHG14 contributes to the progression of NSCLC through miR-206/G6PD pathway
Affiliations
- PMID: 32153123
- PMCID: PMC7180566
- DOI: 10.1111/1759-7714.13374
Item in Clipboard
LncRNA SNHG14 contributes to the progression of NSCLC through miR-206/G6PD pathway
Thorac Cancer.
2020 May.
Abstract
Background: Previous studies have shown that the dysregulation of lncRNAs participates in non-small cell lung cancer (NSCLC) development. The purpose of this study was to research the biological function of lncRNA SNHG14 and its molecular mechanism in NSCLC progression.
Methods: RT-PCR was applied for investigating the expression of SNHG14, miR-206 and G6PD. The progression of NSCLC was detected by CCK-8, Transwell and western blot assays. The targets of SNHG14 and miR-206 were measured by dual-luciferase reporter assay.
Results: We found a higher expression of SNHG14 in NSCLC and upregulation of SNHG14 contributed to NSCLC cell proliferation, invasion and migration. However, knockdown of SNHG14 showed the opposite effect on the progression of NSCLC. Specifically, SNHG14 negatively regulated miR-206 expression by binding with it directly. Furthermore, G6PD served as the target of miR-206. Rescue experiments showed that SNHG14 promoted G6PD expression by inhibiting miR-206.
Conclusions: LncRNA SNHG14 contributed to NSCLC progression through miR-206/G6PD axis, providing novel clues for understanding the mechanism of NSCLC.
Keywords: G6PD; NSCLC; lncRNA SNHG14; miR-206.
© 2020 The Authors. Thoracic Cancer published by China Lung Oncology Group and John Wiley & Sons Australia, Ltd.
Figures
SNHG14 was increased in NSCLC. (a) There was a higher expression of SNHG14 in NSCLC tissues than in nontumor tissues. (b) Increased expression of SNHG14 in NSCLC cells (A549 and SK‐MES‐1 cells) in comparison with that in normal 16HBE cells. *P < 0.05.
SNHG14 modulated NSCLC progression. (a) Measurement of SNHG14 expression in two NSCLC cells after overexpression or silencing SNHG14. (
) NC siRNA, (
) SNHG14 siRNA, (
) NC vector and (
) SNHG14 vector. (b) Cell viability. A549 (
) SNHG14 vector, (
) NC vector, (
) SNHG14 siRNA and (
) NC siRNA. (c) Cell viability. SK‐MES‐1 (
) SNHG14 vector, (
) NC vector, (
) SNHG14 siRNA and (
) NC siRNA. (d) Cell migration. (
) NC siRNA, (
) SNHG14 siRNA, (
) NC vector and (
) SNHG14 vector. (e) Cell invasion. (
) NC siRNA, (
) SNHG14 siRNA, (
) NC vector and (
) SNHG14 vector. *P < 0.05, **P < 0.01.
) NC siRNA, () SNHG14 siRNA, () NC vector and () SNHG14 vector. (b) Cell viability. A549 () SNHG14 vector, () NC vector, () SNHG14 siRNA and () NC siRNA. (c) Cell viability. SK‐MES‐1 () SNHG14 vector, () NC vector, () SNHG14 siRNA and () NC siRNA. (d) Cell migration. () NC siRNA, () SNHG14 siRNA, () NC vector and () SNHG14 vector. (e) Cell invasion. () NC siRNA, () SNHG14 siRNA, () NC vector and () SNHG14 vector. *P < 0.05, **P < 0.01.
SNHG14 acted as a sponge for miR‐206 in NSCLC cells. (a) The prediction binding sites of SNHG14 and miR‐206 are shown. (b, c) The luciferase activity detected in A549 and SK‐MES‐1 cells with a different transfection of miR‐206 mimic, miR‐206 inhibitor, con mimic, con inhibitor. (
) con mimic, (
) miR‐206 mimic, (
) con inhibitor and (
) miR‐206 inhibitor. (
) con mimic, (
) miR‐206 mimic, (
) con inhibitor and (
) miR‐206 inhibitor. (d) The expression level of miR‐206 measured in NSCLC tissues. (e) The mRNA level of miR‐206 tested in the two NSCLC cells with different transfections of NC siRNA, SNHG14 siRNA, NC vector and SNHG14 vector. (
) NC siRNA, (
) SNHG14 siRNA, (
) NC vector and (
) SNHG14 vector. *P < 0.05, **P < 0.01.
) con mimic, () miR‐206 mimic, () con inhibitor and () miR‐206 inhibitor. () con mimic, () miR‐206 mimic, () con inhibitor and () miR‐206 inhibitor. (d) The expression level of miR‐206 measured in NSCLC tissues. (e) The mRNA level of miR‐206 tested in the two NSCLC cells with different transfections of NC siRNA, SNHG14 siRNA, NC vector and SNHG14 vector. () NC siRNA, () SNHG14 siRNA, () NC vector and () SNHG14 vector. *P < 0.05, **P < 0.01.
SNHG14 modulated NSCLC progression by targeting miR‐206. Two NSCLC cells were transfected with SNHG14 siRNA or vector, combined with miR‐206 mimic. (a and b) Cell viability in NSCLC cells after being treated with different transfections SNHG14 siRNA or vector, combined with miR‐206 mimic. (
) SNHG14 siRNA+miR‐206 mimic, (
) SNHG14 siRNA and (
) control. (
) SNHG14 siRNA+miR‐206 mimic, (
) SNHG14 siRNA and (
) control. (c and d) Cell invasion and (e and f) cell migration in the two NSCLC cells with different transfections of SNHG14 siRNA or vector, combined with miR‐206 mimic. *P < 0.05, **P < 0.01. 
) SNHG14 siRNA+miR‐206 mimic, () SNHG14 siRNA and () control. () SNHG14 siRNA+miR‐206 mimic, () SNHG14 siRNA and () control. (c and d) Cell invasion and (e and f) cell migration in the two NSCLC cells with different transfections of SNHG14 siRNA or vector, combined with miR‐206 mimic. *P < 0.05, **P < 0.01.
G6PD served as the target of miR‐101 in NSCLC cells. (a) MiR‐206 and G6PD are the binding sites. (b, c) The luciferase activity of G6PD 3′UTR‐WT or ‐MUT in A549 and SK‐MES‐1 cells after being treated with different transfections of miR‐206 mimic, miR‐206 inhibitor, con mimic, con inhibitor. A549 (
) con mimic, (
) miR‐206 mimic, (
) con inhibitor and (
) miR‐206 inhibitor. SK‐MES‐1 (
) con mimic, (
) miR‐206 mimic, (
) con inhibitor and (
) miR‐206 inhibitor. (d, e) The protein level and mRNA level of G6PD tested in the two NSCLC cells with different transfections. (
) con mimic, (
) miR‐206 mimic, (
) con inhibitor and (
) miR‐206 inhibitor. *P < 0.05, **P < 0.01.
) con mimic, () miR‐206 mimic, () con inhibitor and () miR‐206 inhibitor. SK‐MES‐1 () con mimic, () miR‐206 mimic, () con inhibitor and () miR‐206 inhibitor. (d, e) The protein level and mRNA level of G6PD tested in the two NSCLC cells with different transfections. () con mimic, () miR‐206 mimic, () con inhibitor and () miR‐206 inhibitor. *P < 0.05, **P < 0.01.
G6PD served as the downstream target of SNHG14 in NSCLC. (a) G6PD mRNA. (
) Control, (
) SNHG14 vector and (
) SNHG14 vector+miR‐206 mimic. (b) G6PD protein level detected in the two NSCLC cells with different transfections of SNHG14 vector and miR‐206 mimic. (
) Control, (
) SNHG14 siRNA and (
) SNHG14 siRNA+miR‐206 mimic. (c) The relationship between G6PD and SNHG14 expression. (d) The relationship between G6PD and miR‐206 expression. *P < 0.05, **P < 0.01.
) Control, () SNHG14 vector and () SNHG14 vector+miR‐206 mimic. (b) G6PD protein level detected in the two NSCLC cells with different transfections of SNHG14 vector and miR‐206 mimic. () Control, () SNHG14 siRNA and () SNHG14 siRNA+miR‐206 mimic. (c) The relationship between G6PD and SNHG14 expression. (d) The relationship between G6PD and miR‐206 expression. *P < 0.05, **P < 0.01.Similar articles
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