4t1 murine mammary cancer cells  (Keygen Biotech)

Journal: Biomolecules and Biomedicine

Article Title: Decitabine suppresses tumor growth by activating mouse mammary tumor virus and interferon-β pathways

doi: 10.17305/bb.2025.12852

Figure Lengend Snippet: DAC treatment, tumor growth/metastasis assessments, and MMTV expression readouts in murine tumor models. (A) Tumor growth curves for 4T1 (BALB/c) and MC38 (C57BL/6) tumors treated with decitabine (DAC) or vehicle (PBS); n ═ 5 mice per group. Individual trajectories are shown with group medians overlaid ( x -axis: Days after inoculation; y -axis: Tumor volume, mm 3 ); (B) Tumor growth rates derived as linear-regression slopes of the curves in (A). Points indicate group medians with 95% CIs; P values are annotated; (C) Final tumor mass across strains and treatments: 4T1 in BALB/c and NU/J; MC38 in C57BL/6 and NU/J. n ═ 5 per group except 4T1 in NU/J ( n ═ 9). Medians with 95% CIs; (D) Lung metastasis of 4T1 tumors quantified as log10 colony counts from lung cell cultures. n ═ 5 for BALB/c; n ═ 9 for NU/J. Medians with 95% CIs; (E) MMTV env and pol RNA in 4T1 tumors (BALB/c) measured by qRT-PCR and displayed as log10 relative quantity (PBS n ═ 6; DAC n ═ 5); (F) MMTV env and pol RNA in MC38 tumors (C57BL/6) measured by qRT-PCR ( n ═ 5 per group); (G) Immunoblot of MMTV Env in 4T1 tumors: Lanes 1–5 PBS; lanes 6–10 DAC. Bands near ∼80 kDa and ∼45 kDa are shown; GAPDH (37 kDa) is the loading control. Bar plot summarizes band intensities per lane; (H) Scatter plot of tumor mass versus Env band intensity for the 10 4T1 tumors in (G); five PBS and five DAC samples. Spearman correlation statistics are annotated in the panel. Abbreviations: DAC: Decitabine (5-aza-2ʹ-deoxycytidine); PBS: Phosphate-buffered saline; MMTV: Mousemammary tumor virus; env/pol: MMTV genes; qRT-PCR: Quantitative reverse-transcription PCR; kDa: Kilodalton; CI: Confidence interval; BALB/c = B/c; C57BL/6 = B/6; NU/J: Nude mice; n: Number of mice.

Article Snippet: The 4T1 murine mammary cancer cell line (American Type Culture Collection; STR authenticated) was cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS).

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Western Blot, Control, Saline, Virus, Reverse Transcription

Journal: Biomolecules and Biomedicine

Article Title: Decitabine suppresses tumor growth by activating mouse mammary tumor virus and interferon-β pathways

doi: 10.17305/bb.2025.12852

Figure Lengend Snippet: MMTV knockdown reduces sensitivity of 4T1 cells and tumors to DAC treatment. (A) Effect of MMTV knockdown and DAC treatment on proliferation of 4T1 cells. Cell viability was determined by NucleoCounter analysis after DAC exposure; (B) Effect of MMTV knockdown on DAC-mediated growth inhibition of 4T1 cells in vitro . Percent inhibition was calculated relative to untreated controls; (C) Impact of MMTV knockdown and DAC on tumor growth kinetics of 4T1 tumors in BALB/c mice. Tumor volumes were monitored over time following inoculation and treatment with PBS or DAC; (D) Growth rate analysis of 4T1 tumors in BALB/c mice after MMTV knockdown and DAC treatment. Pairwise comparisons were performed between PBS- and DAC-treated groups, as well as between control and knockdown tumors; (E) Quantification of MMTV env transcript levels in control and knockdown tumors with or without DAC treatment. Relative expression was determined by qRT-PCR; (F) Quantification of MMTV pol transcript levels in control and knockdown tumors with or without DAC treatment. Relative expression was determined by qRT-PCR. Values represent group medians with 95% confidence intervals. n ═ 4 for in vitr o assays (A and B); n ═ 9 for control tumors, n ═ 10 for KD1, and n ═ 5 for KD3 in vivo experiments (C and D); n ═ 4–6 for transcript quantification (E and F). Abbreviations: MMTV: Mouse mammary tumor virus; DAC: Decitabine; PBS: Phosphate-buffered saline; KD: Knockdown; qRT-PCR: Quantitative reverse transcription polymerase chain reaction.

Article Snippet: The 4T1 murine mammary cancer cell line (American Type Culture Collection; STR authenticated) was cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS).

Techniques: Knockdown, Inhibition, In Vitro, Control, Expressing, Quantitative RT-PCR, In Vivo, Virus, Saline, Reverse Transcription, Polymerase Chain Reaction

Journal: Biomolecules and Biomedicine

Article Title: Decitabine suppresses tumor growth by activating mouse mammary tumor virus and interferon-β pathways

doi: 10.17305/bb.2025.12852

Figure Lengend Snippet: IFN-β mediates the antitumor effect of DAC and reciprocally regulates MMTV expression. (A) DAC induces transcriptional upregulation of Ifnb1 in MC38 cells. Expression was quantified by qRT-PCR; (B) DAC treatment increases IFN-β and IRF7 protein levels in 4T1 tumors. BALB/c mice were treated daily for 5 days and every other day thereafter. Protein abundance was analyzed by immunoblotting. Normalized band intensities are shown below the blots; (C) MMTV knockdown reduces IFN-β protein expression in 4T1 cells. Western blot analysis of KD1–KD3 knockdown lines compared with control; (D) Knockdown of Ifnb1 increases MMTV Env expression in 4T1 cells. Western blot showing reciprocal regulation of IFN-β and MMTV Env; (E) Effect of IFN-α on the proliferation of Ifnb1 knockdown cell lines; (F) Effect of Ifnb1 knockdown and DAC treatment on 4T1 cell proliferation. Viable cell counts were determined by NucleoCounter; (G) Ifnb1 knockdown reduces sensitivity of 4T1 cells to DAC in vitro . Percent inhibition was calculated relative to untreated controls; (H) Tumor growth kinetics of control and Ifnb1 knockdown 4T1 tumors in BALB/c mice treated with PBS or DAC. Tumor volumes were measured longitudinally after inoculation; (I) Growth rate analysis of 4T1 tumors following Ifnb1 knockdown and DAC treatment. Pairwise comparisons included untreated and DAC-treated groups, as well as untreated tumors from control vs KD1. Values represent medians with 95% confidence intervals. n ═ 4 for in vitro proliferation assays (A, E–G); n ═ 6–7 for control and KD1, and n ═ 10 for KD2 in vivo studies (H and I). Abbreviations: DAC: Decitabine; IFN: Interferon; IFN-β: Interferon beta; IFN-α: Interferon alpha; IRF7: Interferon regulatory factor 7; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; MMTV: Mouse mammary tumor virus; Env: Envelope protein; KD: Knockdown; PBS: Phosphate-buffered saline; qRT-PCR: Quantitative reverse transcription polymerase chain reaction.

Article Snippet: The 4T1 murine mammary cancer cell line (American Type Culture Collection; STR authenticated) was cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS).

Techniques: Expressing, Quantitative RT-PCR, Quantitative Proteomics, Western Blot, Knockdown, Control, In Vitro, Inhibition, In Vivo, Virus, Saline, Reverse Transcription, Polymerase Chain Reaction