Journal: Oncology Letters

Article Title: Assessment of ALK gene fusions in lung cancer using the differential expression and exon integrity methods

doi: 10.3892/ol.2016.4157

Figure Lengend Snippet: Detection of anaplastic lymphoma kinase fusion in urine from non-small cell lung cancer patients using the differential expression method.

Article Snippet: NSCLC cells and clinical samples The NCI-H3122 human NSCLC cell line (ALK fusion-positive) was provided by AstraZeneca Innovation Center (Shanghai, China).

Techniques: Quantitative Proteomics, Fluorescence In Situ Hybridization, Control

Journal: CytoJournal

Article Title: Oncogene 5’-3’ exoribonuclease 2 enhances epidermal growth factor receptor signaling pathway to promote epithelial–mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.25259/Cytojournal_49_2024

Figure Lengend Snippet: XRN2 was upregulated in NSCLC. (a) The messenger RNA levels of XRN2 in the H460 NSCLC cell line and BEAS-2B human bronchial epithelial cells. (b and c) Protein expression levels of XRN2 in H460 and BEAS-2B cells. n = 6. (*** P < 0.001). (XRN2: 5’-3’ exoribonuclease 2, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, H460 cells: NSCLC cell line; BEAS-2B: Human bronchial epithelial cells, NSCLC: Non-small-cell lung cancer.)

Article Snippet: Human NSCLC cell line H460 (iCell-h160) and normal human bronchial epithelial cells BEAS-2B (iCell-h023) were obtained from iCell Biological Science Company (Shanghai, China).

Techniques: Expressing

Journal: CytoJournal

Article Title: Oncogene 5’-3’ exoribonuclease 2 enhances epidermal growth factor receptor signaling pathway to promote epithelial–mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.25259/Cytojournal_49_2024

Figure Lengend Snippet: XRN2 promoted migration and EMT progression in NSCLC cells. (a-c) Validation of XRN2 overexpression and knockdown efficiency in H460 cells. (d-g) Migration and invasion assay of H460 cells after XRN2 overexpression and knockdown. (h-k) Protein levels of E-cadherin, N-cadherin, and vimentin in H460 cells after XRN2 overexpression and knockdown. n = 6. (** P < 0.01 and *** P < 0.001). (Ov-NC: Overexpress negative control, XRN2: 5’-3’ exoribonuclease 2, Sh-NC: ShRNA negative control, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, H460 cells: NSCLC cell line, BEAS-2B: Human bronchial epithelial cells, NSCLC: Non-small-cell lung cancer.)

Article Snippet: Human NSCLC cell line H460 (iCell-h160) and normal human bronchial epithelial cells BEAS-2B (iCell-h023) were obtained from iCell Biological Science Company (Shanghai, China).

Techniques: Migration, Over Expression, Knockdown, Invasion Assay, Negative Control, shRNA

Journal: CytoJournal

Article Title: Oncogene 5’-3’ exoribonuclease 2 enhances epidermal growth factor receptor signaling pathway to promote epithelial–mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.25259/Cytojournal_49_2024

Figure Lengend Snippet: XRN2 promoted angiogenesis in NSCLC lung metastasis. (a and b) HE stained images of lung metastases. (c and d) IHC analysis of CD31 + cells in lung metastatic lesions. (e and f) Images of tube formation by HUVECs co-cultured with H460-Ov-XRN2 or H460-Sh-XRN2 cells. (g-i) mRNA and protein expression levels of VEGFA in HUVECs under co-culture conditions. n = 6. (** P < 0.01; *** P < 0.001). (Ov-NC: Overexpress negative control, XRN2: 5’-3’ exoribonuclease 2, Sh-NC: ShRNA negative control, CD31: Cluster of differentiation 31, FOV: Field of view, VEGFA: Vascular endothelial growth factor A, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, H460 cells: NSCLC cell line, BEAS-2B: Human bronchial epithelial cells, NSCLC: Non-small-cell lung cancer, Ov-XRN2: XRN2 overexpression, HUVECs: Human umbilical vein endothelial cells .)

Article Snippet: Human NSCLC cell line H460 (iCell-h160) and normal human bronchial epithelial cells BEAS-2B (iCell-h023) were obtained from iCell Biological Science Company (Shanghai, China).

Techniques: Staining, Cell Culture, Expressing, Co-Culture Assay, Negative Control, shRNA, Over Expression

Journal: CytoJournal

Article Title: Oncogene 5’-3’ exoribonuclease 2 enhances epidermal growth factor receptor signaling pathway to promote epithelial–mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.25259/Cytojournal_49_2024

Figure Lengend Snippet: XRN2 overexpression promoted the phosphorylation of EGFR in NSCLC cells. (a) The protein bands of p-EGFR and EGFR. (b) Relative expression levels of the p-EGFR/EGFR protein ratio. (c) The EGFR mRNA levels in H460 cells. n = 6. (*** P < 0.001). (Ov-NC: Overexpress negative control, XRN2: 5’-3’ exoribonuclease 2, Sh-NC: ShRNA negative control, EGFR: Epidermal growth factor receptor, p-EGFR: Phosphorylation epidermal growth factor receptor, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, NSCLC: Non-small-cell lung cancer.)

Article Snippet: Human NSCLC cell line H460 (iCell-h160) and normal human bronchial epithelial cells BEAS-2B (iCell-h023) were obtained from iCell Biological Science Company (Shanghai, China).

Techniques: Over Expression, Expressing, Negative Control, shRNA

Journal: CytoJournal

Article Title: Oncogene 5’-3’ exoribonuclease 2 enhances epidermal growth factor receptor signaling pathway to promote epithelial–mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.25259/Cytojournal_49_2024

Figure Lengend Snippet: EGFR mediated the biological functions of XRN2 in NSCLC metastasis. (a-d) Transwell assays of H460 cells after transfection with Sh-XRN2 and Ov-EGFR. (e-h) The protein expression levels of E-cadherin, N-cadherin, and vimentin in H460 cells post-transfected with Sh-XRN2 and Ov-EGFR. (i and j) Tube formation by HUVECs co-cultured with H460-Sh-XRN2 or H460-Sh-XRN2+Ov-EGFR. n = 6. (* P < 0.05, ** P < 0.01 and *** P < 0.001). (XRN2: 5’-3’ exoribonuclease 2, Sh-NC: ShRNA negative control, Ov-NC: overexpression negative control, EGFR: Epidermal growth factor receptor, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, NSCLC: Non-small cell lung cancer, Ov-EGFR: EGFR overexpression, HUVECs: Human umbilical vein endothelial cells.)

Article Snippet: Human NSCLC cell line H460 (iCell-h160) and normal human bronchial epithelial cells BEAS-2B (iCell-h023) were obtained from iCell Biological Science Company (Shanghai, China).

Techniques: Transfection, Expressing, Cell Culture, shRNA, Negative Control, Over Expression

Journal: Signal Transduction and Targeted Therapy

Article Title: GPR54 regulates non-small cell lung cancer development via dopa decarboxylase

doi: 10.1038/s41392-026-02591-x

Figure Lengend Snippet: GPR54 silencing results in the apoptosis of NSCLC cells. a Western blot analysis of GPR54 expression in normal lung epithelial cells and NSCLC cells. b Xenograft tumor growth curves comparing si Control and si GPR54 groups in NSCLC cell lines ( n = 7/group). Scrambled control siRNAs (si Control ), GPR54 siRNA (si GPR54 ). c Heatmap showing the western blot data. d Quantification of apoptotic cell numbers was measured via TUNEL assays in NSCLC (shown as fold change relative to si Control ). e A retardation of NSCLC cell proliferation by GPR54 gene silencing. f Colony formation by GPR54 silencing. g Annexin V apoptosis analysis after GPR54 silencing. Western blot analysis of phosphorylation levels of both Erk and Akt in mouse lung tissues at 5 weeks ( h ) or at 10 weeks ( i ) after Ad5- CMV-Cre infection. The bar graphs show the relative levels of p-Erk and p-Akt in the mouse lung tissues. j Western blot analysis of apoptotic effect in GPR54 -silenced H358 and HCC1588 cells with HA-Myr-AKT1 overexpression. k Western blot analysis of apoptotic effect in GPR54 -silenced H358 and HCC1588 cells with myc-MEK-ERK2 overexpression. All data present as the mean ± SD. *, p < 0.05; n.s, not significant

Article Snippet: NSCLC cell lines (A549, H460, H1299, H1975, HCC95, HCC1588, H358, and HCC827) were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Western Blot, Expressing, Control, TUNEL Assay, Phospho-proteomics, Infection, Over Expression

Journal: Signal Transduction and Targeted Therapy

Article Title: GPR54 regulates non-small cell lung cancer development via dopa decarboxylase

doi: 10.1038/s41392-026-02591-x

Figure Lengend Snippet: GPR54 is required for aerobic glycolysis in NSCLC cells. ( a ) Venn diagram showing the numbers of differentially expressed genes. ( b ) GSEA/KEGG gene sets were downregulated by oncogenic Kras gene mutation in Gpr54 -null lung tissues compared with wild-type lung tissues. ( c ) Gene enrichment plot of the KEGG_Glycolysis_Gluconeogenesis gene set from the GSEA/KEGG gene sets. (d-i) Seahorse glycolytic stress test in H358 cells after silencing of GPR54 , GNAQ ( Ga q ) , GNA12 ( Ga 12 ) or ARRB2 ( β-arrestin-2 ) followed by stimulation with 100 nM KP10. Oxygen consumption rate (OCR) ( d ), extracellular acidification rate (ECAR) ( e ), proton efflux rate (PER) ( f ), mOCR ( g ), basal GlycoPER ( h ), and compensatory GlycoPER ( i ). j – o Glycolysis stress test in H358 cells pre-treated for 10 minutes with 100 nM KP234 (GPR54 antagonist), 1 μM FR900359 (Gα q/11 inhibitor), 25μM LY294002 (PI3K inhibitor), 1 μM rapamycin (mTOR inhibitor), 10 μM barbadin (β-arrestin inhibitor), 25 mM LiCl (GSK-3β inhibitor), and 25 μM PD98059 (MEK inhibitor), followed by stimulation with 100 nM KP10. OCR ( j ), ECAR ( k ), PER ( l ), mOCR ( m ), basal GlycoPER ( n ), and compensatory GlycoPER ( o ). p Relative glucose consumption levels after GPR54 , GNAQ, GNA12 and ARRB2 silencing, followed by 100 nM KP10 in H358 cells. q Relative lactate production levels after GPR54 , GNAQ, GNA12 and ARRB2 silencing, followed by 100 nM KP10 in H358 cells. r Relative percentages of apoptotic H358 cells, as determined by Annexin V assays, following silencing of GPR54, GNAQ, GNA12 , or ARRB2 . s Western blots for cleaved Caspase-3 and PARP by GNAQ and ARRB2 silencing in H358 and HCC1588 cells. t Relative glucose consumption levels of H358 cells treated with 100 nM KP234, 1 μM FR900359 , 25 μM LY294002, 1 μM rapamycin, 10 μM barbadin, 25 mM LiCl and 25 μM PD98059, followed by 100 nM KP10 10 min later. u Relative lactate production levels in H358 cells treated with 100 nM KP234, 1 μM FR900359 , 25 μM LY294002, 1 μM rapamycin, 10 μM barbadin, 25 mM LiCl and 25 μM PD98059, followed by 100 nM KP10 10 min later. v Relative percentages of apoptotic H358 cells, as determined by Annexin V assays, following treatment with 100 nM KP234, 1 μM FR900359 , 25 μM LY294002, 1 μM rapamycin, 10 μM barbadin, 25 mM LiCl and 25 μM PD98059. w Western blots for cleaved Caspase-3 and PARP by 100 nM KP234, 1 μM FR900359 , 25 μM LY294002, 1 μM rapamycin and 25 μM PD98059 in H358 and HCC1588 cells. All data present the mean ± SD. *, p < 0.05

Article Snippet: NSCLC cell lines (A549, H460, H1299, H1975, HCC95, HCC1588, H358, and HCC827) were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Mutagenesis, Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: GPR54 regulates non-small cell lung cancer development via dopa decarboxylase

doi: 10.1038/s41392-026-02591-x

Figure Lengend Snippet: GPR54-DDC pathway is required for NSCLC cell proliferation. a Differences in protein phosphorylation levels between scrambled control siRNA (si Control ) and DDC siRNA (si DDC ) knockdown A549 cells. b GO analysis of genes significantly altered via DAVID. c Comparison of tumor volume between the si Control group and the si DDC group in a xenograft assay using NSCLC cell lines (each group n = 7). d Relative number of apoptotic cells in tumor tissues from nude mice xenografted with DDC knockdown NSCLC cells was determined via a TUNEL assay. e Growth curves for 96 hours after DDC knockdown in NSCLC cells. f Effects of DDC knockdown on colony formation in NSCLC cells. g Heatmap showing protein levels after DDC silencing in NSCLC cells. h Relative apoptotic cell numbers after DDC knockdown in NSCLC (H358, HCC827, H460 and HCC1588) cells from Annexin V assays. i Bar plot of the levels of phosphorylated proteins most significantly downregulated by DDC knockdown, with a cutoff of a fold change ≤ 0.5 (log₂ ≤ –1) and p < 0.05. Glycolysis stress test in H358 and HCC1588 cells after DDC silencing. OCR ( j ), ECAR ( k ), PER ( l ), mOCR ( m ), basal GlycoPER ( n ), and compensatory GlycoPER ( o ) in H358 cells after DDC silencing, followed by 100 nM KP10 in H358 cells. OCR ( p ), ECAR ( q ), PER ( r ), mOCR ( s ), basal GlycoPER ( t ), and compensatory GlycoPER ( u ) after DDC silencing, followed by 100 nM KP10 in HCC1588 cells. All data present the mean ± SD. *, p < 0.05

Article Snippet: NSCLC cell lines (A549, H460, H1299, H1975, HCC95, HCC1588, H358, and HCC827) were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Phospho-proteomics, Control, Knockdown, Comparison, Xenograft Assay, TUNEL Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: GPR54 regulates non-small cell lung cancer development via dopa decarboxylase

doi: 10.1038/s41392-026-02591-x

Figure Lengend Snippet: Scheme of GPR54-DDC pathway signaling. GPR54 is stimulated by Kisspeptin signals to G αq /G 11 and G β /G γ . The G αq /G 11 pathway activates the PI3K/AKT/mTOR/NF-κB pathway. GPR54-mediated regulation of HK2 expression is crucial for glycolytic function, thereby regulating glucose consumption and lactate production. The G β /G γ pathway activates KRAS/MEK/ERK signaling and/or regulates β-arrestin2-associated ERK signaling to control proliferation. GPR54-dependent NF-κB activation regulates DDC expression, resulting in a positive feedback loop that maintains proliferation and glycolysis. In summary, GPR54 signaling via DDC is crucial for maintaining NSCLC cell proliferation and metabolic reprogramming. BioRender ( www.biorender.com ) was used for the image production

Article Snippet: NSCLC cell lines (A549, H460, H1299, H1975, HCC95, HCC1588, H358, and HCC827) were purchased from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Control, Activation Assay