rabbit antibody against bpgm  (Proteintech)

Journal: Neoplasia (New York, N.Y.)

Article Title: BPGM as an intrinsic brake to constrain metastasis through phospho-epigenetic-mediated carnitine biosynthesis suppression

doi: 10.1016/j.neo.2026.101299

Figure Lengend Snippet: Overexpression of BPGM inhibits tumor metastasis in vitro and in vivo . (A) Silencing BPGM promoted migration of tumor cells. Tumor cells stably expressing shBPGM and its control cells (shCtrl) were examined. Scale bar, 200 μm. (B) Overexpressing BPGM suppressed migration of tumor cells. Tumor cells stably expressing BPGM and its control cells (Ctrl) were examined. Scale bar, 200 μm. (C) The mRNA levels of MMP2 and MMP9 reduced in BPGM-overexpressing tumor cells. SK-HEP-1 cells stably expressing BPGM (BPGM-OE) and its ctrl cells were employed to detect the mRNA levels of MMP2 and MMP9 by qPCR analysis. β-actin was used as an internal control. (D-E) Xenografts of stable Bpgm-overexpressing cells displayed a lower rate of liver and lung metastasis and less metastatic nodules in the liver. For (D), Hepa-Ctrl ( n = 7) and Hepa-BPGM sublines ( n = 6) were inoculated under the capsule of the left hepatic lobe of C57BL/6 mice. Upper panel, a schematic diagram of orthotopic hepatic implantation model. The number of metastatic rate and nodules is shown (D, lower panel). Scale bar, 1 cm. Hematoxylin-eosin staining was performed on serial sections of livers (E, left panel) and lungs (E, right panel) to detect the metastatic nodules. The red arrows indicated the metastatic nodules. Scale bar in left panel, 200 µm; Scale bar in right panel, 100 μm. (F-H) Overexpressing of BPGM inhibited lung metastasis of tumor xenografts. Scale bar in F, 1 cm. B16-F10 cells transfected with Ctrl ( n = 6) or BPGM-OE ( n = 6) was injected into the tail vein of C57BL/6 mice, respectively. Upper panel in F, a schematic diagram of lung metastasis model by tail vein injection. H&E staining of lung sections was performed to observe metastatic foci (G). For G, scale bar in left panel, 500 μm; scale bar in right panel, 200 μm. The number of melanoma nodules and lung metastasis is shown in H. (I) The model deciphers the inhibitory role of BPGM in tumor metastasis. Error bar: mean ± SEM. P -values are labeled above the bar chart.

Article Snippet: The antibodies used included mouse antibody against β-actin (BM0627, Boster, Wuhan, China), rabbit antibody against BPGM (17173-1-AP, Proteintech), EZH2 (F0281, Selleck), phospho-EZH2 (Thr345) (TA3584S, Abmart, Shanghai, China), phospho-CDK1 (Thr14) (AP1465, Abclonal, Wuhan, China), ubiquitin (10201-2-AP, Proteintech), HIF1α (36169, Cell Signaling Technology, CST, Beverly, MA, USA), H3K4me3 (91264, Active Motif), H3K79me3 (cat 49-1020, Thermos Fisher), H3K9me3 (61014, Active Motif), H3K27me3 (91168, Active Motif) and Histone 3 (F0057, Selleck).

Techniques: Over Expression, In Vitro, In Vivo, Migration, Stable Transfection, Expressing, Control, Staining, Transfection, Injection, Labeling

Journal: Neoplasia (New York, N.Y.)

Article Title: BPGM as an intrinsic brake to constrain metastasis through phospho-epigenetic-mediated carnitine biosynthesis suppression

doi: 10.1016/j.neo.2026.101299

Figure Lengend Snippet: BPGM constrains cell migration by decreasing l -carnitine to paralyze β-oxidation. (A) Metabolic flux map from ¹³C₆-glucose tracing. (B-C) The levels of ¹³C₆-glucose derived metabolic intermediates in tumors. 5 % (w/v) of 13 C 6 labeled- d -glucose was injected into mice bearing Hepa-BPGM-OE or Hepa-Ctrl tumors by tail vein for 0.5 hours. The tumor tissues were employed to conduct glucose metabolic flux analysis. The blue dash box indicated the metabolites of RLS. (D) The ratio of ¹³C₆-glucose derived two-carbon labelled lactate ( M + 2) verse three-carbon labelled lactate ( M + 3). (E) The untargeted metabolomics experimental protocol. (F) Volcano plot displayed differentially expressed metabolites in tumor tissue and plasma. (G) GO enrichment analysis showed significant enrichment of methionine synthesis, carnitine synthesis, and fatty acid oxidation pathways in BPGM-overexpressing tumor tissue and plasma. (H) Heatmap showed that l -carnitine and most of acyl-carnitines were downregulated in BPGM-OE group. Columns: individual samples; rows: metabolites. (I) l -carnitine was significantly reduced in BPGM-OE tumor tissues and plasmas. The concentration of l -carnitine was quantified by LC-MS/MS. (J-K) Replenishing l -carnitine attenuated the inhibitory effect of BPGM on the migration of tumor cells. Sublines with stably overexpression of BPGM were treated with 1 mM l -carnitine for 72 hours followed by transwell assays. The total number of migrated cells were stained by violet and counted under microscope. Scale bar, 200 μm. (L) The model deciphered l -carnitine mediated the inhibitory role of BPGM in cellular migration. Error bar: mean ± SEM. P -values are labeled above the bar chart.

Article Snippet: The antibodies used included mouse antibody against β-actin (BM0627, Boster, Wuhan, China), rabbit antibody against BPGM (17173-1-AP, Proteintech), EZH2 (F0281, Selleck), phospho-EZH2 (Thr345) (TA3584S, Abmart, Shanghai, China), phospho-CDK1 (Thr14) (AP1465, Abclonal, Wuhan, China), ubiquitin (10201-2-AP, Proteintech), HIF1α (36169, Cell Signaling Technology, CST, Beverly, MA, USA), H3K4me3 (91264, Active Motif), H3K79me3 (cat 49-1020, Thermos Fisher), H3K9me3 (61014, Active Motif), H3K27me3 (91168, Active Motif) and Histone 3 (F0057, Selleck).

Techniques: Migration, Derivative Assay, Labeling, Injection, Clinical Proteomics, Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Stable Transfection, Over Expression, Staining, Microscopy

Journal: Neoplasia (New York, N.Y.)

Article Title: BPGM as an intrinsic brake to constrain metastasis through phospho-epigenetic-mediated carnitine biosynthesis suppression

doi: 10.1016/j.neo.2026.101299

Figure Lengend Snippet: 2,3-BPG-CDK1-EZH2-H3K27me3 Axis: BPGM’s epigenetic circuit breaker for cellular migration. (A) Integrated functional metabolomics analysis revealed BPGM-altered metabolites clustered in methyl donor group. Bubble size: metabolites count. (B) Hypothesis of molecular mechanism underlying BPGM regulated BBOX1 expression by post transcriptional modification (PTM). (C) Silencing BPGM significantly reduced the protein level of H3K27me3, while overexpressing BPGM increased its level. Cells stably expressing shBPGM/BPGM and its control cells (shCtrl/Ctrl) were used to detect protein level by western blotting. (D) ChIP assays disclosed that the fragments of BBOX1 and MMP9 promoter precipitated by anti-H3K27me3 antibody were increased upon overexpressing BPGM. SK-HEP-1 cells stably expressing BPGM and its control cells (Ctrl) were employed to ChIP assay. The antibody precipitated DNAs were amplified by qPCR. 5 % of the total DNAs were amplified to serve as the control for DNA content. Values shown are signal of α-H3K27me3-precipitated DNA relative to the input and the mean value of the control group was normalized as 1. (E) Overexpressing BPGM significantly increased the protein level of EZH2 but decreased the protein level of p-EZH2-T 345 in tumor cells. (F) The molecular docking of 2,3-BPG and CDK1. Predicted structure of 2,3-BPG binding with CDK1. Key contact residues: Thr14, Arg127, Arg170. (G) Overexpressing BPGM significantly increased the protein level of p-CDK1-T 14 in tumor cells. Cells stably expressing BPGM (BPGM-OE) and its control cells (Ctrl) were used to detect protein level by western blotting. (H) 2,3-BPG treatment enhanced the phosphorylation of CDK1 at thr14 in tumor cells. The indicated concentration of 2,3-BPG was incubated with the lysate of trophoblasts and tumor cells for 30 minutes followed by western blotting. (I-J) RO-3306 treatment enhanced the phosphorylation of CDK1 at thr14 and reduced the phosphorylation of EZH2 at thr345 in tumor cells. The tumor cells were treated with the indicated concentration of RO-3306 for 12 hours followed by western blotting. (K) The model deciphers the role of BPGM in regulating BBOX1 and MMP9 expression. Error bar: mean ± SEM. P -values are labeled above the bar chart.

Article Snippet: The antibodies used included mouse antibody against β-actin (BM0627, Boster, Wuhan, China), rabbit antibody against BPGM (17173-1-AP, Proteintech), EZH2 (F0281, Selleck), phospho-EZH2 (Thr345) (TA3584S, Abmart, Shanghai, China), phospho-CDK1 (Thr14) (AP1465, Abclonal, Wuhan, China), ubiquitin (10201-2-AP, Proteintech), HIF1α (36169, Cell Signaling Technology, CST, Beverly, MA, USA), H3K4me3 (91264, Active Motif), H3K79me3 (cat 49-1020, Thermos Fisher), H3K9me3 (61014, Active Motif), H3K27me3 (91168, Active Motif) and Histone 3 (F0057, Selleck).

Techniques: Migration, Functional Assay, Expressing, Modification, Stable Transfection, Control, Western Blot, Amplification, Binding Assay, Phospho-proteomics, Concentration Assay, Incubation, Labeling

Journal: Neoplasia (New York, N.Y.)

Article Title: BPGM as an intrinsic brake to constrain metastasis through phospho-epigenetic-mediated carnitine biosynthesis suppression

doi: 10.1016/j.neo.2026.101299

Figure Lengend Snippet: Working model of BPGM-mediated metabolic-epigenetic regulation axis and its gatekeeper role in tumor metastasis. In low-metastatic tumors, higher oxygen levels activate KDM4A, which removes repressive H3K9me3 marks at the BPGM promoter, thereby promoting BPGM transcription. Elevated BPGM expression increases the production of 2,3-BPG, which stabilizes EZH2 and enhances SAM-dependent H3K27me3 deposition. This epigenetic remodeling leads to transcriptional silencing of BBOX1 , a key gene involved in carnitine biosynthesis, consequently suppressing fatty acid oxidation and inhibiting tumor metastasis. In contrast, under hypoxic conditions commonly found in high-metastatic tumors, KDM4A activity is diminished, resulting in the accumulation of H3K9me3 at the BPGM promoter and subsequent downregulation of BPGM expression. This disruption of the BPGM-mediated regulatory axis abrogates its anti-metastatic function. Notably, preclinical studies revealed that pharmacological inhibition of BBOX1 with Meldonium restores the metabolic-epigenetic barrier, effectively impeding metastatic progression.

Article Snippet: The antibodies used included mouse antibody against β-actin (BM0627, Boster, Wuhan, China), rabbit antibody against BPGM (17173-1-AP, Proteintech), EZH2 (F0281, Selleck), phospho-EZH2 (Thr345) (TA3584S, Abmart, Shanghai, China), phospho-CDK1 (Thr14) (AP1465, Abclonal, Wuhan, China), ubiquitin (10201-2-AP, Proteintech), HIF1α (36169, Cell Signaling Technology, CST, Beverly, MA, USA), H3K4me3 (91264, Active Motif), H3K79me3 (cat 49-1020, Thermos Fisher), H3K9me3 (61014, Active Motif), H3K27me3 (91168, Active Motif) and Histone 3 (F0057, Selleck).

Techniques: Expressing, Activity Assay, Disruption, Inhibition