Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: NRG1 expression is elevated in RAW264.7 macrophages co-cultured with 4T1 cells. (A) The expression of NRG1 was examined using next-generation RNA sequencing analysis. (B) The mRNA expression of NRG1 in RAW264.7 macrophages co-cultured with 4T1 cells was detected using reverse transcription-quantitative PCR. (C) The protein expression of NRG1 in RAW264.7 macrophages co-cultured with 4T1 cells was detected using immunoblotting analysis. *** P<0.001. NRG1, neuregulin 1.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Cell Culture, RNA Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Hypoxic MDA-MB-231 cells induces the M2 polarization of THP-1 macrophages. (A) The protein expression of HIF-1α was assessed using immunoblotting analysis. (B) The protein expression of HIF-1α in hypoxic MDA-MB-231/THP-1 co-cultures was assessed using immunoblotting analysis. (C) The cell migration was detected using Transwell. (D and E) The level of CD163 was detected using flow cytometry. The levels of (F) IL-10 and (G) IL-12 were detected using ELISA-related assay kits. ** P<0.01 and *** P<0.001. HIF-1α, hypoxia-inducible factor-1α.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Western Blot, Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: NRG1 expression is associated with M2 polarization and poor prognosis. (A) The expression of NRG1 in BC samples using TCGA database. (B) Spearman correlation analysis between NRG1 and CD163 expression in breast cancer samples. (C) The association of NRG1 expression with the overall survival in BC patients. NRG1, neuregulin 1; BC, breast cancer; TCGA, The Cancer Genome Atlas; GEPIA, Gene Expression Profiling Interactive Analysis.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Gene Expression

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Hypoxic MDA-MB-231 cells induces the M2 polarization of THP-1 macrophages via NRG1. (A) The mRNA expression of NRG1 was detected using RT-qPCR. (B) The transfection efficacy of Ov-NRG1 was detected using RT-qPCR and immunoblotting analysis. (C) The transfection efficacy of si-NRG1 was detected using RT-qPCR and immunoblotting analysis. (D) The level of IL-10 was detected using ELISA-related IL-10 assay kits. (E and F) The level of CD163 was detected using flow cytometry. (G) Following the treatment of anti-NRG1 blocking antibody, the level of IL-10 was detected using ELISA-related IL-10 assay kits. (H-I) Following the treatment of anti-NRG1 blocking antibody, the level of CD163 was detected using flow cytometry. * P<0.05, ** P<0.01 and *** P<0.001. NRG1, neuregulin 1; RT-qPCR, reverse transcription-quantitative PCR; Ov, overexpression; si, small interfering.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Blocking Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Over Expression

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Hypoxic MDA-MB-231 cells mediate the M2 polarization of THP-1 macrophages via exosomes. (A) The cell migration was detected using Transwell. Scale: 100 μ m. (B) The level of CD163 was detected using flow cytometry analysis. (C) The level for IL-10 was detected using ELISA-related IL-10 assay kits. (D) Transmission electron microscope was used for the inspection of exosomes. Scale: 100 μ m. (E) The analysis of exosome particle size. (F) The expressions of exosomal marker proteins were detected using immunoblotting analysis. (G) PKH67 staining was used to trace the exosomes. (H) The level of CD163 was detected using flow cytometry analysis. ** P<0.01 and *** P<0.001.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Transmission Assay, Microscopy, Marker, Western Blot, Staining

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Exosomal CAMTA1 promotes the M2 polarization of THP-1 macrophages. (A) The expression of CAMTA1 in BC samples was predicted using TCGA database. (B) The mRNA expression of CAMTA1 in MDA-MB-231 cells was detected using RT-qPCR. (C) The mRNA expression of CAMTA1 in exosomes was detected using RT-qPCR. (D) The mRNA expression of CAMTA1 in THP-1 macrophages was detected using RT-qPCR. (E) The transfection efficacy of sh-CAMTA1 was detected using RT-qPCR and immunoblotting analysis. (F) The mRNA expression of CAMTA1 in transfected MDA-MB-231 cells was detected using RT-qPCR. (G) The mRNA expression of CAMTA1 in transfected exosomes was detected using RT-qPCR. (H) The cell migration was detected using Transwell. (I and J) The level of CD163 was detected using flow cytometry analysis. (K) The level of IL-10 was detected using ELISA-related IL-10 assay kits. * P<0.05, ** P<0.01 and *** P<0.001. CAMTA1, Calmodulin-binding Transcription Activator 1; BC, breast cancer; TCGA, The Cancer Genome Atlas; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Migration, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Binding Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Exosomal CAMTA1 promotes tumor growth in vivo . (A) The transfection efficacy of Ov-CAMTA1 was detected using RT-qPCR and immunoblotting analysis. (B) The appearance of tumor. The tumor (C) volume and (D) weight. (E) The mRNA expression of CAMTA1 was detected using RT-qPCR. (F) The level of IL-10 was detected using ELISA-related IL-10 assay kits. (G) The level of CD163 was detected using immunohistochemistry analysis. (H) H&E staining. (I) The expression of Caspase 3 was detected using immunohistochemistry analysis. (J) The Spearman correlation analysis of CAMTA1 and NRG1. (K) The expression of NRG1 was detected using immunohistochemistry analysis. * P<0.05, ** P<0.01 and *** P<0.001. CAMTA1, Calmodulin-binding Transcription Activator 1; Ov, overexpression; RT-qPCR, reverse transcription-quantitative PCR; H&E, hematoxylin and eosin; NRG1, neuregulin 1.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: In Vivo, Transfection, Quantitative RT-PCR, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Binding Assay, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction