Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: EphA2 derived from EphA2 High cell exosomes can be internalized by EphA2 Low cells. ( A ) Expression levels of EphA2 in four GC cell lines, n = 3. ( B ) TEM characterization of exosome morphology (scale bar = 100 nm). ( C ) Particle size distribution of exosomes analyzed by NTA. ( D ) Western blot detection of exosomal markers (positive: Alix and CD9; negative: calnexin) in SGC7901 cells and exosomes. ( E ) Localization of exogenous exosomal EphA2 in MGC803 (scale bar = 25 μm). ( F ) Expression levels of EphA2 in MGC803 cells following treatment with exosomes derived from different cell sources, n = 3. (* p < 0.05, *** p < 0.001).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Exosomal EphA2 derived from EphA2 High cells transforms EphA2 Low cells to promote GC progression in vitro. ( A ) Proliferation capacity of SGC7901 cells following treatment with shCtrl- or shEphA2-derived exosomes, analyzed by CCK8 assay, n = 3. ( B ) Proliferation capacity of SGC7901 cells following treatment with shCtrl- or shEphA2-derived exosomes, detected by colony formation assay, n = 3. ( C ) Migration and invasion abilities of SGC7901 cells following treatment with shCtrl- or shEphA2-derived exosomes, detected by transwell assay, n = 3 (scale bar = 200 μm). ( D ) Proliferation capacity of MGC803 cells following treatment with shCtrl- or oeEphA2-derived exosomes, analyzed by CCK8 assay, n = 3. ( E ) Proliferation capacity of MGC803 cells following treatment with shCtrl- or oeEphA2-derived exosomes, detected by colony formation assay, n = 3. ( F ) Migration and invasion abilities of MGC803 cells following treatment with shCtrl- or oeEphA2-derived exosomes, detected by transwell assay, n = 3 (scale bar = 200 μm). (ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay, In Vitro, CCK-8 Assay, Colony Assay, Migration, Transwell Assay

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Exosomal EphA2 derived from EphA2 High cells transforms EphA2 Low cells to promote GC progression in vivo. ( A ) Subcutaneous tumor formation in the left and right axillae of nude mice after injection with oeCtrl- or oeEphA2-derived exosomes ( n = 3 mice). ( B ) Comparison of tumor volume between the two groups. ( C ) Comparison of tumor weight between the two groups. ( D ) Schematic diagram of the experimental design for tail-vein injection of oeCtrl- or oeEphA2-derived exosomes in nude mice ( n = 3 mice). ( E ) In vivo imaging to detect metastatic signals in nude mice. ( F ) In vivo imaging to detect visceral metastatic foci in nude mice. ( G ) H&E staining to detect the formation of lung and liver metastatic foci after injection with oeCtrl- or oeEphA2-derived exosomes (scale bar = 100 μm). (* p < 0.05).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay, In Vivo, Injection, Comparison, In Vivo Imaging, Staining

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Exosomal EphA2-mediated transformation of EphA2 Low to EphA2 High cells promotes GC progression through ligand-independent pathways. ( A ) Western blot analysis of EFNA1 silencing after siRNA transfection. ( B ) Transwell assay to detect changes in the migration ability of MGC803 cells after the addition of shCtrl or shEphA2 exosomes in the context of EFNA1 silencing, n = 3 (scale bar = 200 μm). ( C ) Western blot detection of the impact of exosome addition on the AKT signaling pathway. ( D ) Effect of exosome addition on the ERK signaling pathway, n = 3. ( E ) Expression of p-ERK after the addition of the ERK inhibitor U0126, n = 3. ( F ) CCK8 assay to assess the proliferation ability of MGC803 cells treated with U0126 in the presence of exosomes, n = 3. ( G ) Colony formation assay to evaluate the colony-forming ability of MGC803 cells treated with U0126 in the presence of exosomes, n = 3. ( H ) Transwell assay to measure the migration and invasion abilities of MGC803 cells treated with U0126 in the presence of exosomes, n = 3 (scale bar = 200 μm). ( I ) Expression levels of MMP2 and MMP9 in SGC7901 and MGC803 cells after EphA2 knockdown or overexpression, n = 3. ( J ) Expression levels of p-ERK and MMP2 in MGC803 cells after exosome treatment, n = 3. ( K ) Expression level of MMP2 in SGC7901 exosomes after EphA2 knockdown. (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Transformation Assay, Western Blot, Transfection, Transwell Assay, Migration, Expressing, CCK-8 Assay, Colony Assay, Knockdown, Over Expression

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Correlation between EphA2 expression in serum exosomes of patients with GC and tumor metastasis. ( A ) Overall survival of GC patients with high EphA2 expression ( n = 118). ( B ) Detection of EphA2 expression in human serum exosomes by ELISA ( n = 9). (* p < 0.05).

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Exosomal EphA2 Promotes Gastric Cancer Progression by Inducing Phenotypic Transformation of Tumor Cells in a Ligand-Independent Manner

doi: 10.3390/cells15141253

Figure Lengend Snippet: Schematic model illustrating how exosomal EphA2 derived from EphA2 High cells transforms EphA2 Low cells to promote gastric cancer progression in a ligand-independent manner. The figure was Created in BioRender. Zhan, L. (2026) https://BioRender.com/lsuww4f .

Article Snippet: The sources of antibodies were: EphA2 (Santa Cruz, sc-398832), GAPDH (ZSGB-BIO, Beijing, China, TA-08), Alix (Invitrogen, Waltham, MA, USA, MA1-83977), CD9 (Invitrogen, Bothell, WA, USA, 10626D), CD63 (Invitrogen, Bothell, WA, USA, 10628D), ERK (Bioss, Beijing, China, bsm-2637R), p-ERK (Zenbio, Chengdu, China, R24245 ), AKT (Immunoway, Suzhou, China, YT0185), p-AKT (Cell Signaling Technology, Waltham, MA, USA, 31957), Flag (ABclonal, Wuhan, China, AE005), EFNA1 (ABclonal, Wuhan, China, A9132), p-EphA2-S897 (ABclonal, Wuhan, China, AP1082), p-EphA2-Y772 (ABclonal, Wuhan, China, AP0817).

Techniques: Derivative Assay

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: MMP-9 , a factor that promotes Vasculogenic mimicry, is highly expressed in CC and is associated with poor prognosis. (A) CC database of TCGA was used to analyze key factors associated with VM. (B) Association of Sox2 expression with overall survival in CC (log-rank test). (C) Association of MMP-9 expression with overall survival in CC (log-rank test). (D) Panoramic scans after immunohistochemical detection of MMP-9 and H&E staining in samples from cancerous and paracancerous tissues from subjects with CC. Scale bar, 50 µm. Original magnification, ×20. (E) Protein levels of MMP-9 in 20 paired samples, with the MMP-9 level in CC tissue expressed compared with that in the paired normal tissue. (F) Expression levels of MMP-9 mRNA in 44 paired CC and paracancerous tissues, with MMP-9 expression in CC tissue expressed compared with that in the paired normal tissue. (G) Comparison of the average expression levels of MMP-9 mRNA in CC tissues compared with paracancerous tissues. (H) HeLa and SiHa cells were incubated under hypoxia (0.1% O 2 ) and proteins collected at 24, 48 and 72 h for western blotting of ALDH1, EPHA2, MMP-9 and GAPDH. ImageJ was used to semi-quantify western blotting signals from HeLa (I) and SiHa (J) cells. GAPDH served as an internal reference. *P<0.05, **P<0.01 and ***P<0.001. MMP-9, matrix metalloproteinase 9; VM, vasculogenic mimicry; ALDH1, aldehyde dehydrogenase 1; EPHA2, ephrin type-A receptor 2; TCGA, The Cancer Genome Atlas; Sox2, SRY-box transcription factor 2; CC, cervical cancer; CESC, cervical squamous cell carcinoma.

Article Snippet: The membranes were blocked with 5% milk at 20±5°C for 1 h. The blocked membranes were incubated at 4°C overnight with the following antibodies: A rabbit monoclonal anti-NSUN2 antibody (1:1,000; cat. no. AB259941; Abcam), a rabbit monoclonal anti-transfer RNA aspartic acid methyltransferase 1 (TRDMT1) antibody (1:1,000; cat. no. 19221-1-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology), a rabbit polyclonal anti-MMP-9 antibody (1:1,000; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology), a rabbit polyclonal anti-aldehyde dehydrogenase 1 (ALDH1) antibody (1:1,000; cat. no. 15910-1-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology), a rabbit polyclonal anti-ephrin type-A receptor 2 (EPHA2) antibody (1:1,000; cat. no. AF5 238; Affinity Biosciences) and a rabbit polyclonal anti-GAPDH antibody (1:1,000; TA309157 OriGene Technologies, Inc.).

Techniques: Expressing, Immunohistochemical staining, Staining, Comparison, Incubation, Western Blot