aml cell lines thp 1  (ATCC)

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: Effects of SFXN3 Knockdown on Proliferation, Apoptosis, and Signaling Pathways in AML Cells. (A–C) qRT-PCR and Western blot analyses were used to measure SFXN3 expression levels in various leukemia cell lines (THP-1, KG-1, U937, K562) and in normal bone marrow stromal cells (HS-5). (D) Two independent shRNAs (sh-SFXN3–1 and sh-SFXN3–2) were used to knock down SFXN3 expression in THP-1 and KG-1 cells. Western blot was performed to assess the knockdown efficiency and specificity. (E) Quantification of SFXN3 knockdown efficiency by different shRNAs. (F) CCK-8 cell proliferation assays were conducted to evaluate the effects of SFXN3 knockdown on cell growth dynamics over time. (G) EdU incorporation assays were used to assess DNA synthesis activity, indirectly reflecting cellular proliferation, and to compare differences between knockdown and control groups, (bar=50ųm). (H) Western blot analysis of key cell cycle regulatory proteins (CDK4, CDK6, P27, and P21) to investigate the potential mechanism by which SFXN3 affects cell cycle progression. (I) TUNEL assays were used to evaluate apoptosis levels in the knockdown versus control groups, assessing the role of SFXN3 in apoptosis suppression, (bar=50ųm). (J) Western blot analysis of pro-apoptotic proteins (BAX and BAK) and anti-apoptotic proteins (Bcl-2 and Bcl-xl) in THP-1 and KG-1 cells following SFXN3 knockdown. (K) Correlation analysis between SFXN3 expression and key proteins in the Wnt/β-Catenin signaling pathway. (L) Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. Data are presented as mean ± SD. from three independent experiments ( n = 3). One-way ANOVA was used in (A, B, E), and two-way ANOVA was used in (F). *, p < 0.05.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Knockdown, Protein-Protein interactions, Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, DNA Synthesis, Activity Assay, Control, TUNEL Assay, Fractionation, Translocation Assay, Marker

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: The Wnt/β-Catenin Pathway Agonist SKL2001 Reverses the Effects of SFXN3 Knockdown on Leukemia Cell Proliferation and Apoptosis. (A) Western blot analysis of SFXN3 protein expression following SFXN3 knockdown and treatment with SKL2001, to assess whether SKL2001 significantly modulates SFXN3 expression. (B) CCK-8 assays were performed to evaluate whether SKL2001 could reverse the inhibitory effects of SFXN3 knockdown on the proliferation of THP-1 and KG-1 leukemia cells. (C) EdU staining assays were used to assess DNA synthesis activity, analyzing the ability of SKL2001 to restore proliferation suppressed by SFXN3 knockdown, (bar=50 ųm). (D) Quantitative analysis of EdU fluorescence intensity to evaluate DNA replication across different treatment groups. (E) Western blot analysis of cell cycle regulators CDK4, CDK6, Cyclin D1, and Cyclin E1 to determine whether SKL2001 rescues the expression of these proteins in SFXN3-silenced cells. (F) Western blot analysis of pro-apoptotic proteins (BAX, BAK) and anti-apoptotic proteins (Bcl-2, Bcl-xl) to confirm that SKL2001 mitigates the apoptosis-promoting effects of SFXN3 knockdown. (G) TUNEL assays were conducted to assess whether SKL2001 suppresses the enhanced apoptosis induced by SFXN3 knockdown, (bar=50ųm). (H) Quantification of TUNEL fluorescence intensity, reflecting apoptosis levels under different treatment conditions. (I) Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. Data are presented as mean ± SD. from at least three independent experiments. One-way ANOVA was used in (D, H), and two-way ANOVA was used in (B). *, p < 0.05; **, p < 0.01; ***, p < 0.001 vs. control or scramble group.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Knockdown, Western Blot, Expressing, CCK-8 Assay, Staining, DNA Synthesis, Activity Assay, Fluorescence, TUNEL Assay, Fractionation, Translocation Assay, Marker, Control

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: The REST–SFXN3 Axis Promotes Malignant Phenotypes in AML Cells via the Wnt/β-Catenin Signaling Pathway. (A) Western blot analysis of the effect of REST knockdown (sh-REST) on SFXN3 expression, and the reversal of this effect by SFXN3 overexpression. (B) CCK-8 assays assess the impact of sh-REST and SFXN3 overexpression on AML cell proliferation. (C) EdU incorporation assays evaluate the effects of sh-REST and SFXN3 overexpression on DNA synthesis activity in AML cells, (bar=50ųm). (D) Quantification of EdU-positive cells to compare DNA synthesis capacity across groups. (E) Western blot analysis of proliferation-related proteins CDK4, CDK6, Cyclin D1, and Cyclin E1 under sh-REST and SFXN3 overexpression conditions. (F) Band intensities were quantified using ImageJ software and normalized to the indicated internal controls. (G) TUNEL assays detect apoptotic cells after REST knockdown and SFXN3 overexpression, (bar=50ųm). (G) Quantitative analysis of apoptotic cells in THP-1 and KG-1 cell lines. (H) Quantification of TUNEL fluorescence intensity, reflecting apoptosis levels under different treatment conditions. (I) Western blot evaluation of pro-apoptotic proteins (BAX, BAK) and anti-apoptotic proteins (Bcl-2, Bcl-xl), demonstrating REST knockdown promotes apoptosis, which is reversed by SFXN3 overexpression. (J) Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. Data are presented as mean ± SD from three independent experiments ( n = 3).One-way ANOVA was used in (D, F,H), and two-way ANOVA was used in (B). **, p < 0.01; ***, p < 0.001.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Western Blot, Knockdown, Expressing, Over Expression, CCK-8 Assay, DNA Synthesis, Activity Assay, Software, TUNEL Assay, Fluorescence, Fractionation, Translocation Assay, Marker

Journal: Translational Oncology

Article Title: REST-driven upregulation of SFXN3 promotes AML progression via Wnt/β-catenin activation and confers decitabine resistance

doi: 10.1016/j.tranon.2026.102705

Figure Lengend Snippet: Decitabine Suppresses AML Cell Proliferation and Promotes Apoptosis via SFXN3 Inhibition. (A) RT-PCR analysis of the effects of Gefitinib, Disulfiram, and Decitabine on SFXN3 mRNA expression. (B) Western blot analysis of SFXN3 protein levels following treatment with Gefitinib, Disulfiram, and Decitabine. (C) CCK-8 assay to calculate the IC50 values of Decitabine in THP-1 and KG-1 cells, identifying appropriate drug concentrations for subsequent experiments (D) CCK-8 assays were performed to evaluate AML cell viability at 6,12,24,48, and 72 h following treatment with 50 nm decitabine, thereby determining the optimal treatment duration. E) EdU incorporation assay evaluating the proliferation capacity of AML cells after Decitabine treatment, (bar=50ųm). (F) Western blot analysis of proliferation-related proteins (P21, P27, CDK4, and CDK6) following Decitabine treatment. (G) TUNEL staining to detect DNA fragmentation at the 3′-OH ends, marking apoptotic cells after Decitabine exposure, (bar=50ųm). (H) Western blot analysis of pro-apoptotic (e.g., BAX, BAK) and anti-apoptotic (e.g., Bcl-2, Bcl-xl) protein expression in response to Decitabine. (I) Western blot analysis of key components of the Wnt/β-Catenin signaling pathway after Decitabine treatment, revealing pathway inhibition. Subcellular fractionation followed by Western blotting was performed to assess β-catenin nuclear translocation. Cytoplasmic (Cyto) and nuclear (Nuc) fractions were probed for β-catenin, with β-actin (cytoplasmic marker) and Histon H3 (nuclear marker) used to confirm fractionation quality. n = 3,Error bars indicate mean ± SD; One-way ANOVA in (D, F); **, p < 0.01, *** p <0.001.

Article Snippet: The ATCC supplied the AML cell lines THP-1, KG-1, U937, and K562, and the stromal cell line HS-5.

Techniques: Inhibition, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, CCK-8 Assay, TUNEL Assay, Staining, Fractionation, Translocation Assay, Marker

Journal: Journal of Cell Communication and Signaling

Article Title: Mechanistic role of GNE‐987 targeting BRD4‐HCP5 axis in pediatric T‐cell acute lymphoblastic leukemia

doi: 10.1002/ccs3.70063

Figure Lengend Snippet: BRD4 inhibitor significantly promotes HCP5 super‐enhancer activity and expression. (A) Reverse transcription‐quantitative polymerase chain reaction analysis of HCP5 mRNA expression levels in acute myeloid leukemia cell lines (THP‐1 and HL‐60) treated with the BRD4 inhibitor GNE‐987. (B) Western Blot analysis showing BRD4 expression levels in different groups. (C) Representative agarose gel electrophoresis image and statistical quantification of ChIP‐qPCR products. Compared to the Ctrl group, * p < 0.05, ** p < 0.01. All cell experiments were repeated three times. BRD4, Bromodomain‐containing protein 4; HCP5, HLA Complex P5.

Article Snippet: The acute myeloid leukemia (AML) cell lines Tsuchiya Human Phagocyte‐1 (THP‐1) and Human Leukemia‐60 (HL‐60) (TIB‐202 and CCL‐240, ATCC), along with T‐ALL–specific cell lines Jurkat, CCRF‐CEM, MOLT‐4, and RPMI‐8402 (TIB‐152, CCL‐111, CRL‐1582, and CCL‐27, ATCC), were cultured in RPMI‐1640 medium (11875093, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (A5670701, Gibco) and maintained in an incubator at 37°C with 5% CO 2 .

Techniques: Activity Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Agarose Gel Electrophoresis, ChIP-qPCR

Journal: Journal of Cell Communication and Signaling

Article Title: Mechanistic role of GNE‐987 targeting BRD4‐HCP5 axis in pediatric T‐cell acute lymphoblastic leukemia

doi: 10.1002/ccs3.70063

Figure Lengend Snippet: Bromodomain‐containing protein 4 Inhibitor GNE‐987 Significantly Inhibits acute myeloid leukemia Cell Proliferation and Migration and Induces Apoptosis. (A) CCK‐8 assay measuring cell proliferation in each group at 0, 12, 24, 36, 48, 60, and 72 h, with absorbance detected at OD450. (B) Representative images of Live and Dead staining for each group, with a bar chart depicting the statistical analysis of cell death ratios. Bar = 50 μm. (C) Colony formation assay for each group, with a bar chart showing the statistical analysis of colony numbers. (D) Flow cytometry analysis of apoptosis levels in each group, with a bar chart depicting the statistical analysis of apoptosis rates. Compared with the Ctrl group, * p < 0.05, ** p < 0.01. All cell experiments were repeated three times.

Article Snippet: The acute myeloid leukemia (AML) cell lines Tsuchiya Human Phagocyte‐1 (THP‐1) and Human Leukemia‐60 (HL‐60) (TIB‐202 and CCL‐240, ATCC), along with T‐ALL–specific cell lines Jurkat, CCRF‐CEM, MOLT‐4, and RPMI‐8402 (TIB‐152, CCL‐111, CRL‐1582, and CCL‐27, ATCC), were cultured in RPMI‐1640 medium (11875093, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (A5670701, Gibco) and maintained in an incubator at 37°C with 5% CO 2 .

Techniques: Migration, CCK-8 Assay, Staining, Colony Assay, Flow Cytometry

Journal: Journal of Cell Communication and Signaling

Article Title: Mechanistic role of GNE‐987 targeting BRD4‐HCP5 axis in pediatric T‐cell acute lymphoblastic leukemia

doi: 10.1002/ccs3.70063

Figure Lengend Snippet: Silencing of HLA Complex P5 Reverses the Inhibitory Effect of GNE‐987 on acute myeloid leukemia Cell Viability. (A) Reverse transcription‐quantitative polymerase chain reaction results verify the silencing efficiency of sh‐HCP5. (B) CCK8 assay measuring cell proliferation at 0, 12, 24, 36, 48, 60, and 72 h, with absorbance detected at OD450. (C) Representative images of Live and Dead staining in each group and a bar chart showing the death ratio; scale bar = 50 μm. (D) Colony formation assay and statistical graph of colony numbers for each group. (E) Flow cytometry analysis of apoptosis levels and a statistical graph of apoptosis rates in each group. For panel A, compared with the sh‐NC group, * p < 0.05, ** p < 0.01. For panels B‐E, compared with the Ctrl group, * p < 0.05, ** p < 0.01; compared with the GNE‐987+sh‐NC group, # p < 0.05, ## p < 0.01. All cell experiments were repeated three times.

Article Snippet: The acute myeloid leukemia (AML) cell lines Tsuchiya Human Phagocyte‐1 (THP‐1) and Human Leukemia‐60 (HL‐60) (TIB‐202 and CCL‐240, ATCC), along with T‐ALL–specific cell lines Jurkat, CCRF‐CEM, MOLT‐4, and RPMI‐8402 (TIB‐152, CCL‐111, CRL‐1582, and CCL‐27, ATCC), were cultured in RPMI‐1640 medium (11875093, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (A5670701, Gibco) and maintained in an incubator at 37°C with 5% CO 2 .

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, CCK-8 Assay, Staining, Colony Assay, Flow Cytometry

Journal: Translational Cancer Research

Article Title: High TERF2 expression is associated with poor prognosis and its suppression attenuates progression in acute myeloid leukemia

doi: 10.21037/tcr-2025-1226

Figure Lengend Snippet: Downregulation of TERF2 suppresses AML cells viability and proliferation. (A) qRT-PCR assays analyzed TERF2 mRNA levels of NC or TERF2-knockdown MOLM13 cells and THP1 cells. (B) Western blot assays detected TERF2 protein levels of NC or TERF2-knockdown MOLM13 cells and THP1 cells. (C,D) CCK-8 assay showing the viability of NC or TERF2-knockdown MOLM13 cells (C) and THP1 cells (D). (E-H) Flow cytometry assay analyzed cell cycle in NC or TERF2-knockdown MOLM13 cells (E,F) and THP1 cells (G,H). **, P<0.01; ****, P<0.0001; ns, not significant. AML, acute myeloid leukemia; CCK-8, Cell Counting Kit-8; NC, negative control; qRT-PCR, quantitative reverse transcription polymerase chain reaction.

Article Snippet: AML cell lines (MOLM13 and THP-1) were obtained from the American Type Culture Collection (ATCC) and were cultured in RPMI-1640 medium (Gibco, Grand Island, USA) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco) under standard conditions (37 °C, 5% CO 2 ).

Techniques: Quantitative RT-PCR, Knockdown, Western Blot, CCK-8 Assay, Flow Cytometry, Cell Counting, Negative Control, Reverse Transcription, Polymerase Chain Reaction

Journal: Translational Cancer Research

Article Title: High TERF2 expression is associated with poor prognosis and its suppression attenuates progression in acute myeloid leukemia

doi: 10.21037/tcr-2025-1226

Figure Lengend Snippet: TERF2 deficiency promotes apoptosis in AML cells. (A-D) Flow cytometry assay analyzed apoptosis in NC or TERF2-knockdown MOLM13 cells (A,B) or THP1 cells (C,D). (E-H) Western blot detected cleaved caspase 3 levels of NC or TERF2-knockdown MOLM13 cells (E,F) or THP1 cells (G,H). ***, P<0.001; ****, P<0.0001. AML, acute myeloid leukemia; NC, negative control.

Article Snippet: AML cell lines (MOLM13 and THP-1) were obtained from the American Type Culture Collection (ATCC) and were cultured in RPMI-1640 medium (Gibco, Grand Island, USA) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco) under standard conditions (37 °C, 5% CO 2 ).

Techniques: Flow Cytometry, Knockdown, Western Blot, Negative Control

Journal: Translational Cancer Research

Article Title: High TERF2 expression is associated with poor prognosis and its suppression attenuates progression in acute myeloid leukemia

doi: 10.21037/tcr-2025-1226

Figure Lengend Snippet: TERF2 involved in cuproptosis through E2F pathway in AML. (A) Bar graphs illustrate the associations between TERF2 expression levels and gene hallmark sets in AML patients. (B) GSEA for AML patients with high TERF2 expression in TCGA database. (C-E) Western blot analysis of E2F1, CDK4, CDK6, CDKN2A levels in MOLM13 cells and NB4 cells with or without TERF2 silence. (F,G) MOLM13 (F) and NB4 (G) cells with TERF2 knockdown were treated with specified concentrations of ES-Cu (1:1 ratio) for 48 hours, followed by cell viability assessment using the CCK-8 assay. ***, P<0.001; ****, P<0.0001. AML, acute myeloid leukemia; CCK-8, Cell Counting Kit-8; ES-Cu, elesclomol-copper; GSEA, gene set enrichment analysis; IC 50 , half-maximal inhibitory concentration; TCGA, The Cancer Genome Atlas.

Article Snippet: AML cell lines (MOLM13 and THP-1) were obtained from the American Type Culture Collection (ATCC) and were cultured in RPMI-1640 medium (Gibco, Grand Island, USA) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco) under standard conditions (37 °C, 5% CO 2 ).

Techniques: Expressing, Western Blot, Knockdown, CCK-8 Assay, Cell Counting, Concentration Assay

Journal: Translational Cancer Research

Article Title: High TERF2 expression is associated with poor prognosis and its suppression attenuates progression in acute myeloid leukemia

doi: 10.21037/tcr-2025-1226

Figure Lengend Snippet: Knockdown of TERF2 suppresses AML progression and enhances cuproptosis sensitivity in vivo . (A,B) Tumor growth was monitored by bioluminescence imaging in MOLM13-engrafted NSG mice (A) and the quantification of luciferase signals for all mice per group (B). (C) Kaplan-Meier analysis of survival of MOLM13-engrafted mice, log rank test. (D,E) Tumor growth was monitored by bioluminescence imaging in MOLM13-engrafted mice treated with elesclomol (D) and the quantification of luciferase signals for all mice per group (E). (F) Kaplan-Meier analysis of survival of MOLM13-engrafted mice treated with elesclomol, log rank test. **, P<0.01. AML, acute myeloid leukemia; NSG, NOD-SCID IL2rg.

Article Snippet: AML cell lines (MOLM13 and THP-1) were obtained from the American Type Culture Collection (ATCC) and were cultured in RPMI-1640 medium (Gibco, Grand Island, USA) containing 10% heat-inactivated fetal bovine serum (FBS; Gibco) under standard conditions (37 °C, 5% CO 2 ).

Techniques: Knockdown, In Vivo, Imaging, Luciferase