Journal: Cancer Medicine

Article Title: Iron protects childhood acute lymphoblastic leukemia cells from methotrexate cytotoxicity

doi: 10.1002/cam4.2982

Figure Lengend Snippet: Iron a tiny molecule with huge effects. The schematic diagram demonstrating at least two pathways by which iron may confer resistance to leukemic cells. First, iron induces moderate levels of ROS, shown in gray, through the Fenton reaction mediated by hydrogen peroxide (Fe 2+ + H 2 O 2 → Fe 3+ + OH − + ºOH). ROS, then, may trigger the antioxidant defense system via upregulation of BCL2 and NRF2 . Thereby, the malignant cell can survive against the toxic levels of MTX‐induced ROS (demonstrated in red). At the same time, the antiapoptotic activity of BCL2 may inhibit the MTX cytotoxic effects. Upregulation of BCL2 may also be mediated by iron itself through the overexpression of STAT3 , possibly via IL‐6 and β‐catenin. Moreover, iron enhances MRP1 expression indirectly via the activation of its transcription factor, NRF2. The MTX efflux through this ABC transporter protects the cell from MTX cytotoxicity. , activation; , inhibition/repression; , possible/indirect activation; , transport; , activates transcription of the gene (shown by yellow rectangles); , MRP1 transporter; NRF2, nuclear factor erythroid 2‐related factor 2; MRP1, multidrug resistance‐associated protein 1 (also known as ABCC1); TCF, T‐cell factor/lymphoid enhancing factors family of DNA‐binding factors; AREs, antioxidant response elements; RR, ribonucleotide reductase; MTX, methotrexate; ROS, reactive oxygen species

Article Snippet: Methotrexate (MTX) was bought from Santa Cruz Biotechnology, Inc. Deferasirox (Exjade) (DFX) was obtained from Novartis.

Techniques: Activity Assay, Over Expression, Expressing, Activation Assay, Inhibition, Binding Assay

Journal: Toxicology

Article Title: A 3D spheroid model of quadruple cell co-culture with improved liver functions for hepatotoxicity prediction.

doi: 10.1016/j.tox.2024.153829

Figure Lengend Snippet: Fig. 6 Drug-induced lipid dysregulation replicated by the 3D quadruple cell co- culture liver spheroid model. (A) Neutral fat visualization and quantification in the spheroid model after treatment of isoniazid, palmitic acid, nifedipine, or amiodarone via BODIPY™ 493/503 staining (n=3 for palmitic acid treatment, n=4 for other compounds). (B) Phospholipid imaging and quantification in the isoniazid or amiodarone-treated spheroid model through NBD-PE labeling (n=4). Scale bar: 200 μm. *p < 0.05, **p < 0.01, compared to the control group.

Article Snippet: In the test of drug-induced liver fibrosis risk, methotrexate (TMstandard, China) was adopted as the tested compound, and 48-hour TGF-β1 (MedChemExpress) treatment at a concentration of 10 ng/mL during Day 4-6 was selected for setting up the positive control.

Techniques: Co-Culture Assay, Staining, Imaging, Labeling, Control

Journal: Nature Communications

Article Title: Metabolic control of CD47 expression through LAT2-mediated amino acid uptake promotes tumor immune evasion

doi: 10.1038/s41467-022-34064-4

Figure Lengend Snippet: a Immunohistochemistry analysis of CD47 expression (CD47 H-score) in matched osteosarcoma specimens from the same patient before and after chemotherapy ( n = 81 patients). Scale bar, 400 μm. b , c Cisplatin or doxorubicin was injected intravenously every 2 days, and ifosfamide or methotrexate was injected intravenously every 4 days, after GFP + HOS tumor grew for 14 days. b Western blot analysis of CD47 expression in HOS tumors on day 22. c Flow cytometric analysis of CD47 protein expression in GFP + HOS cells from tumors on day 22 (left; n = 4 mice per group). The anti-CD47 median fluorescence intensity (MFI) was determined (right). d Kaplan–Meier curves of overall survival (OS) of osteosarcoma patients stratified by CD47 expression in pre-chemotherapy specimens. High CD47 expression (CD47 high ): CD47 H-score > 0.9; low CD47 expression (CD47 low ): CD47 H-score ≤ 0.9. e , f ΔCD47 H-score for each patient was obtained from subtracting CD47 H-score of pre-chemotherapy specimen from that of paired post-chemotherapy specimen. e Kaplan–Meier curves of overall survival (OS) of osteosarcoma patients stratified by ΔCD47 expression. High ΔCD47 expression (ΔCD47 high ): ΔCD47 H-score > 0.8; low ΔCD47 expression (ΔCD47 low ): ΔCD47 H-score ≤ 0.8. f Kaplan–Meier curves of overall survival (OS) of osteosarcoma patients stratified by CD47 expression in pre-chemotherapy specimens and ΔCD47 expression. Low CD47 expression and low ΔCD47 expression (CD47 low ΔCD47 low ): CD47 H-score ≤ 0.9 and ΔCD47 H-score ≤ 0.8; low CD47 expression and high ΔCD47 expression (CD47 low ΔCD47 high ): CD47 H-score ≤ 0.9 and ΔCD47 H-score > 0.8; high CD47 expression and low ΔCD47 expression (CD47 high ΔCD47 low ): CD47 H-score > 0.9 and ΔCD47 H-score ≤ 0.8; high CD47 expression and high ΔCD47 expression (CD47 high ΔCD47 high ): CD47 H-score > 0.9 and ΔCD47 H-score > 0.8. Data are shown as the mean ± SD. ns, not significant. * P < 0.05, ** P < 0.01, *** P < 0.001, paired two-tailed Student t test ( a ), one-way ANOVA ( c ) or log-rank test ( d – f ). The experiment was performed three times with similar results ( b , c ). See the Source Data file for the exact P values. Source data are provided as a Source Data file.

Article Snippet: Ifosfamide (240 mg/kg; Selleck Chemicals) and methotrexate disodium (240 mg/kg, Selleck Chemicals) were administrated intravenously three times every 4 days.

Techniques: Immunohistochemistry, Expressing, Injection, Western Blot, Fluorescence, Two Tailed Test