Journal: Cells

Article Title: The Warburg Trap: A Novel Therapeutic Approach for Targeting Osteosarcoma.

doi: 10.3390/cells13010061

Figure Lengend Snippet: Figure 2. Warburg Trap drugs significantly reduce the colony-forming capacity of OS cells. OS cell lines (n = 6) were seeded at low density in culture plates and treated with the WT drugs (Metformin (1 mM), Monensin (5 nM), and Fenofibrate (10 µM)) in an acidic culture medium (pH 6.5). After 24 h and 48 h of treatment, the culture medium was replaced by a normal cell culture medium with a physiological pH value of 7.4. Colonies that formed during a further seven days of culture were stained with haematoxylin and photographed before (A) the number of colonies and (B) the size of the colonies were quantified using the software ImageJ version 1.48v (* p < 0.01 compared to untreated controls). (C) Representative photographs of stained colonies obtained with cell line 143B analysed in duplicates. (D) Analysis of colony numbers that formed after pre-treatment of osteosarcoma cell lines (n = 6) with WT drugs and subsequent cultivation for 14 days in soft agar (* p < 0.01 compared to untreated controls).

Article Snippet: The following six commercially available human osteosarcoma cell lines were used in this study: HOS (American Tissue Culture Collection, Rockville, MD, USA), HOS 143B (Sigma-Aldrich, Taufkirchen, Germany), CAL-72 (DSMZ, Braunschweig, Germany), MNNG-HOS, SaOS-2, and U2OS (Cell Line Service GmbH, Eppelheim, Germany).

Techniques: Cell Culture, Staining, Software

Journal: Cells

Article Title: The Warburg Trap: A Novel Therapeutic Approach for Targeting Osteosarcoma.

doi: 10.3390/cells13010061

Figure Lengend Snippet: Figure 3. DDIT3-mediated downregulation of WNT target genes by WT drugs. Osteosarcoma cell lines (n = 3) were treated with WT drugs (Metformin (1 mM), Monensin (5 nM), and Fenofibrate (10 µM)) (MeMoFe) for the indicated time before the expression of DDIT3 and the WNT target genes AXIN2, CCND1 and LGR5 were analysed by real-time quantitative RT-PCR analysis. The expression of the reference gene RPS13 (ribosomal protein S13) was used for normalisation. Data are presented as percent of untreated 24 h controls (* p < 0.05 compared to untreated controls).

Article Snippet: The following six commercially available human osteosarcoma cell lines were used in this study: HOS (American Tissue Culture Collection, Rockville, MD, USA), HOS 143B (Sigma-Aldrich, Taufkirchen, Germany), CAL-72 (DSMZ, Braunschweig, Germany), MNNG-HOS, SaOS-2, and U2OS (Cell Line Service GmbH, Eppelheim, Germany).

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Translational Medicine

Article Title: SLC38A5 suppresses ferroptosis through glutamine-mediated activation of the PI3K/AKT/mTOR signaling in osteosarcoma

doi: 10.1186/s12967-024-05803-6

Figure Lengend Snippet: SLC38A5 is up-regulated in osteosarcoma and associated with a poor prognosis. ( A ) The expression patterns of SLC38A5 across various tumors in the TCGA dataset based on the TIMER database. ( B ) The relative mRNA level of SLC38A5 in tumor ( n = 12) and adjacent normal tissues ( n = 12) based on clinical samples. ( C ) The protein level of SLC38A5 was assessed in osteosarcoma tissues and adjacent normal counterparts using western blot. ( D ) The quantitative analysis of the western blot result. ( E-F ) The protein level of SLC38A5 in human osteoblastic cell (hFOB1.19) and osteosarcoma cell lines (HOS, Saos-2, MG63, U2OS, 143B). ( G ) The mRNA level of SLC38A5 in different cell lines. (H-I) Kaplan–Meier survival analysis of SLC38A5 in osteosarcoma based on the TARGET database ( n = 86) and GSE21257 dataset ( n = 53). The median value of SLC38A5 expression is the cut-off for high and low expression. Results from three independent experiments are summarized in a histogram format. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The human osteosarcoma cell lines HOS, Saos-2 and MG63 were purchased from Wuhan Servicebio Technology Co., Ltd.

Techniques: Expressing, Western Blot