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Ru3 promoted ROS accumulation by inducing mitochondrial damage in cisplatin-resistant lung cancer cells A Mitochondrial morphology was captured in cells with and without the addition of 5 μM Ru3 using transmission electron microscopy. Scale bar, 2 μm. B Mitotracker-Green was utilized to assess mitochondrial mass, independent of mitochondrial membrane potential (MMP) alterations, in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. The analysis was conducted using a fluorescence microplate reader (Thermo Scientific). C JC-1 analysis was performed to assess mitochondrial membrane potential changes in A549-DDP and PC9-DDP cells in the presence or absence of 5 μM Ru3. D The number of mitochondrial DNA copies was examined by qRT-PCR in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. E qRT-PCR analysis was performed to detect MRPL40, MRPL35, MRPL14, GCDH, and RBP1 in PC9-DDP cells after treatment with 5 μM Ru3 for 24 h. F – H The colocalization of Mitotracker Red and DCFH-DA was analyzed in PC9-DDP cells both with and without the treatment of 5 μM Ru3. Scale bar, 10 μm. I – J MitoSOX Red was used to evaluate oxide levels in mitochondria of A549-DDP and PC9-DDP cells upon treatment with 5 μM Ru3. Scale bar, 10 μm. K ROS levels were determined by flow cytometry in A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 <t>μM</t> <t>Mito-TEMPO</t> for 24 h. L – M The cell viability of A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h or 48 h was detected by CCK-8 assays. (N) Lactate dehydrogenase release was analyzed in PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 48 h. Data are representative of three independent experiments and are presented as means ± s.d. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Statistical significance was determined by comparing with the respective control groups
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Ru3 promoted ROS accumulation by inducing mitochondrial damage in cisplatin-resistant lung cancer cells A Mitochondrial morphology was captured in cells with and without the addition of 5 μM Ru3 using transmission electron microscopy. Scale bar, 2 μm. B Mitotracker-Green was utilized to assess mitochondrial mass, independent of mitochondrial membrane potential (MMP) alterations, in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. The analysis was conducted using a fluorescence microplate reader (Thermo Scientific). C JC-1 analysis was performed to assess mitochondrial membrane potential changes in A549-DDP and PC9-DDP cells in the presence or absence of 5 μM Ru3. D The number of mitochondrial DNA copies was examined by qRT-PCR in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. E qRT-PCR analysis was performed to detect MRPL40, MRPL35, MRPL14, GCDH, and RBP1 in PC9-DDP cells after treatment with 5 μM Ru3 for 24 h. F – H The colocalization of Mitotracker Red and DCFH-DA was analyzed in PC9-DDP cells both with and without the treatment of 5 μM Ru3. Scale bar, 10 μm. I – J MitoSOX Red was used to evaluate oxide levels in mitochondria of A549-DDP and PC9-DDP cells upon treatment with 5 μM Ru3. Scale bar, 10 μm. K ROS levels were determined by flow cytometry in A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 <t>μM</t> <t>Mito-TEMPO</t> for 24 h. L – M The cell viability of A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h or 48 h was detected by CCK-8 assays. (N) Lactate dehydrogenase release was analyzed in PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 48 h. Data are representative of three independent experiments and are presented as means ± s.d. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Statistical significance was determined by comparing with the respective control groups
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Ru3 promoted ROS accumulation by inducing mitochondrial damage in cisplatin-resistant lung cancer cells A Mitochondrial morphology was captured in cells with and without the addition of 5 μM Ru3 using transmission electron microscopy. Scale bar, 2 μm. B Mitotracker-Green was utilized to assess mitochondrial mass, independent of mitochondrial membrane potential (MMP) alterations, in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. The analysis was conducted using a fluorescence microplate reader (Thermo Scientific). C JC-1 analysis was performed to assess mitochondrial membrane potential changes in A549-DDP and PC9-DDP cells in the presence or absence of 5 μM Ru3. D The number of mitochondrial DNA copies was examined by qRT-PCR in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. E qRT-PCR analysis was performed to detect MRPL40, MRPL35, MRPL14, GCDH, and RBP1 in PC9-DDP cells after treatment with 5 μM Ru3 for 24 h. F – H The colocalization of Mitotracker Red and DCFH-DA was analyzed in PC9-DDP cells both with and without the treatment of 5 μM Ru3. Scale bar, 10 μm. I – J MitoSOX Red was used to evaluate oxide levels in mitochondria of A549-DDP and PC9-DDP cells upon treatment with 5 μM Ru3. Scale bar, 10 μm. K ROS levels were determined by flow cytometry in A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 <t>μM</t> <t>Mito-TEMPO</t> for 24 h. L – M The cell viability of A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h or 48 h was detected by CCK-8 assays. (N) Lactate dehydrogenase release was analyzed in PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 48 h. Data are representative of three independent experiments and are presented as means ± s.d. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Statistical significance was determined by comparing with the respective control groups
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Ru3 promoted ROS accumulation by inducing mitochondrial damage in cisplatin-resistant lung cancer cells A Mitochondrial morphology was captured in cells with and without the addition of 5 μM Ru3 using transmission electron microscopy. Scale bar, 2 μm. B Mitotracker-Green was utilized to assess mitochondrial mass, independent of mitochondrial membrane potential (MMP) alterations, in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. The analysis was conducted using a fluorescence microplate reader (Thermo Scientific). C JC-1 analysis was performed to assess mitochondrial membrane potential changes in A549-DDP and PC9-DDP cells in the presence or absence of 5 μM Ru3. D The number of mitochondrial DNA copies was examined by qRT-PCR in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. E qRT-PCR analysis was performed to detect MRPL40, MRPL35, MRPL14, GCDH, and RBP1 in PC9-DDP cells after treatment with 5 μM Ru3 for 24 h. F – H The colocalization of Mitotracker Red and DCFH-DA was analyzed in PC9-DDP cells both with and without the treatment of 5 μM Ru3. Scale bar, 10 μm. I – J MitoSOX Red was used to evaluate oxide levels in mitochondria of A549-DDP and PC9-DDP cells upon treatment with 5 μM Ru3. Scale bar, 10 μm. K ROS levels were determined by flow cytometry in A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 <t>μM</t> <t>Mito-TEMPO</t> for 24 h. L – M The cell viability of A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h or 48 h was detected by CCK-8 assays. (N) Lactate dehydrogenase release was analyzed in PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 48 h. Data are representative of three independent experiments and are presented as means ± s.d. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Statistical significance was determined by comparing with the respective control groups
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Ru3 promoted ROS accumulation by inducing mitochondrial damage in cisplatin-resistant lung cancer cells A Mitochondrial morphology was captured in cells with and without the addition of 5 μM Ru3 using transmission electron microscopy. Scale bar, 2 μm. B Mitotracker-Green was utilized to assess mitochondrial mass, independent of mitochondrial membrane potential (MMP) alterations, in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. The analysis was conducted using a fluorescence microplate reader (Thermo Scientific). C JC-1 analysis was performed to assess mitochondrial membrane potential changes in A549-DDP and PC9-DDP cells in the presence or absence of 5 μM Ru3. D The number of mitochondrial DNA copies was examined by qRT-PCR in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. E qRT-PCR analysis was performed to detect MRPL40, MRPL35, MRPL14, GCDH, and RBP1 in PC9-DDP cells after treatment with 5 μM Ru3 for 24 h. F – H The colocalization of Mitotracker Red and DCFH-DA was analyzed in PC9-DDP cells both with and without the treatment of 5 μM Ru3. Scale bar, 10 μm. I – J MitoSOX Red was used to evaluate oxide levels in mitochondria of A549-DDP and PC9-DDP cells upon treatment with 5 μM Ru3. Scale bar, 10 μm. K ROS levels were determined by flow cytometry in A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h. L – M The cell viability of A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h or 48 h was detected by CCK-8 assays. (N) Lactate dehydrogenase release was analyzed in PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 48 h. Data are representative of three independent experiments and are presented as means ± s.d. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Statistical significance was determined by comparing with the respective control groups

Journal: Apoptosis

Article Title: Ru3: a novel ruthenium-based compound overcomes cisplatin resistance in non-small cell lung cancer via the E2F2-IGFBP3-GPX4 pathway

doi: 10.1007/s10495-026-02447-3

Figure Lengend Snippet: Ru3 promoted ROS accumulation by inducing mitochondrial damage in cisplatin-resistant lung cancer cells A Mitochondrial morphology was captured in cells with and without the addition of 5 μM Ru3 using transmission electron microscopy. Scale bar, 2 μm. B Mitotracker-Green was utilized to assess mitochondrial mass, independent of mitochondrial membrane potential (MMP) alterations, in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. The analysis was conducted using a fluorescence microplate reader (Thermo Scientific). C JC-1 analysis was performed to assess mitochondrial membrane potential changes in A549-DDP and PC9-DDP cells in the presence or absence of 5 μM Ru3. D The number of mitochondrial DNA copies was examined by qRT-PCR in A549-DDP and PC9-DDP cells treated with or without 5 μM Ru3 for 24 h. E qRT-PCR analysis was performed to detect MRPL40, MRPL35, MRPL14, GCDH, and RBP1 in PC9-DDP cells after treatment with 5 μM Ru3 for 24 h. F – H The colocalization of Mitotracker Red and DCFH-DA was analyzed in PC9-DDP cells both with and without the treatment of 5 μM Ru3. Scale bar, 10 μm. I – J MitoSOX Red was used to evaluate oxide levels in mitochondria of A549-DDP and PC9-DDP cells upon treatment with 5 μM Ru3. Scale bar, 10 μm. K ROS levels were determined by flow cytometry in A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h. L – M The cell viability of A549-DDP and PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 24 h or 48 h was detected by CCK-8 assays. (N) Lactate dehydrogenase release was analyzed in PC9-DDP cells treated with 5 μM Ru3 in the presence or absence of 10 μM Mito-TEMPO for 48 h. Data are representative of three independent experiments and are presented as means ± s.d. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Statistical significance was determined by comparing with the respective control groups

Article Snippet: NAC (HY-B0215), Mito-TEMPO (HY-112879), HLM006474 (HY-16667), Z-VAD-FAK (HY-16658B), irinotecan (HY-13001), docetaxel (HY-13065), Necrostatin-1 (HY-15760), NLRP3-IN-78 (HY-144623), 4-PBA (HY-100457), Fer-1 (HY-100579), RSL3 (HY-100218) were purchased from MCE.

Techniques: Morphology, Transmission Assay, Electron Microscopy, Membrane, Analysis, Fluorescence, Quantitative RT-PCR, Flow Cytometry, CCK-8 Assay, Control