Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Antitumor effect of organometallic compounds in osteosarcoma cell lines. Cell viability (WST-1 assay) in 2D cultures measured after the treatment of 143B Par (A), OS833 Par (B), 143B CisPt-R (C), and OS833 CisPt-R (D) with increasing concentrations of free or encapsulated drugs for 72 h. The effect of equivalent amounts of empty nanoparticles in each cell line is shown. IC 50 values for each treatment are indicated. Error bars represent the standard deviation of three independent experiments. Asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon) (**** p < 0.0001; two-way ANOVA Tukey's test). (E and F) Analysis of DNA damage (formation of −H2AX foci) in 143B CisPt-R cells treated with the IC 75 of CisPt (8.4 µM), Ru3 (0.9 µM), LIP-Ru3 (0.8 µM), or the equivalent amount of empty nanoparticles (LIP-Empty) for 24 hours. Representative images of immunofluorescence staining (scale bar = 25 µm) (E) and quantification of the formation of gH2AX foci in >100 cells (F) are shown. Asterisks indicate statistically significant differences (** p < 0.01; *** p < 0.001; One-way ANOVA Tukey's test). (G) Cell cycle distribution of 143B CisPt-R cells treated with the IC 50 of CisPt (5 µM), Ru3 (0.5 µM), LIP-Ru3 (0.5 µM), or the equivalent amount of empty nanoparticles (LIP-Empty) for 48 hours. The normalized percentage of cells in each cell cycle phase (mean ± standard deviation of three independent experiments) is shown. Asterisks indicate statistically significant differences in G 0 /G 1 phase (blue) between LIP-empty control and Ru3-formulations and in the G 2 /M phase (pink) between LIP-empty and CisPt (**** p < 0.0001; two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: WST-1 Assay, Standard Deviation, Immunofluorescence, Staining, Control

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Cytotoxic effect of organometallic compounds in 3D parental spheroids. Spheroid viability (CTG 3D assay) was measured after the treatment of 143B Par (A and B) and OS833 Par (C and D) spheroids with increasing concentrations of free and encapsulated drugs for 96 h. The effect of equivalent amounts of empty nanoparticles is shown. Representative images of 143B Par (A) and OS833 Par (C) spheroids treated with increasing concentrations of each compound are shown (scale bars = 250 µm). Likewise, cell survival curves and IC50 values (µM) for each condition in 143B Par (B) and OS833 Par (D) spheroids are presented as the mean ± the standard deviation of at least eight biological replicates. Asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon) (**** p < 0.0001; two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: Standard Deviation

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Cytotoxic effect of organometallic compounds in 3D cisplatin-resistant spheroids. Spheroid viability (CTG 3D assay) was measured after the treatment of 143B CisPt-R (A and B) and OS833 CisPt-R (C and D) spheroids with increasing concentrations of free and encapsulated drugs for 96 h. The effect of equivalent amounts of empty nanoparticles is shown. Representative images of 143B CisPt-R (A) and OS833 CisPt-R (C) spheroids treated with increasing concentrations of each compound are shown (scale bars = 250 µm). Cell survival curves and IC50 values (µM) for each condition in 143B CisPt-R (B) and OS833 CisPt-R (D) are presented as the mean ± standard deviation of at least eight biological replicates. Asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon) (* p < 0.05; ** p < 0.01; **** p < 0.0001; two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: Standard Deviation

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: Effect of free and nano-encapsulated Ru3 in CSC-enriched 3D osteosarcoma cell cultures. 143B Par cells were plated at low density in tumorsphere medium and left to form tumorspheres for 9 days before being treated for 96 h with increasing concentrations of Ru3, LIP-Ru3, and cisplatin. Treatments with the same amount of empty nanoparticles (LIP-Empty) were also included. Representative images (scale bars = 250 μm) (A), quantification of the number of tumorspheres remaining after the treatment (B), and cell viability (WST-1 assay) (C) are presented. Data is presented as the mean ± the standard deviation of independent replicates. In panel B, black asterisks indicate statistically significant differences between controls and the indicated conditions in each treatment. In panel C, asterisks indicate statistically significant differences between cisplatin and Ru3 (black) or LIP-Ru3 (salmon); (** p < 0.01; *** p < 0.001; one-way (panel B) or two-way (panel C) ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: WST-1 Assay, Standard Deviation

Journal: Drug Delivery

Article Title: Liposome-mediated delivery of a ruthenium-based metallodrug to overcome cisplatin resistance in osteosarcoma

doi: 10.1080/10717544.2026.2671485

Figure Lengend Snippet: In vivo antitumor effect of free and nanoencapsulated metallic compounds. Established 143B CisPt-R xenografts were randomly assigned to six different groups ( n = 5 per group) and treated i.v. with vehicle (PBS, control), empty liposomes (LIP-Empty), free Ru3 (4 mg/kg, Ru3), ruthenium-loaded liposomes (4 mg/kg, LIP-Ru3), RAPTA-C (4 mg/kg), and cisplatin (4 mg/kg) once a week for 3 weeks (cumulative dose of 12 mg/kg). (A) Curves representing the mean relative tumor volume of 143B CisPt-R xenografts during the treatments. Drug efficacy expressed as the percentage of TGI is indicated. (B) Average tumor weight (left panel) and images of the tumors (right panel) at the end of the experiment (day 20 after the start of the treatment). (C) Change in the body weights of mice during the treatments. Error bars represent the SEM and asterisks indicate statistically significant differences between groups (* p < 0.05; **** p < 0.0001, two-way ANOVA Tukey's test).

Article Snippet: Human osteosarcoma cell line 143B (CRL-8303) was obtained from the ATCC repository (Manassas, VA, USA).

Techniques: In Vivo, Control, Liposomes

Journal: Cell Discovery

Article Title: ASB7 promotes osteosarcoma lung metastasis through ubiquitin-mediated degradation of ATF2

doi: 10.1038/s41421-026-00890-9

Figure Lengend Snippet: a The relative copy number, mRNA expression of ASB7 and cell line count of each tumor type were plotted using 25Q2 DepMap cancer cell line data. b RNA-seq analysis of ASB7 mRNA levels in normal ( n = 4) and osteosarcoma ( n = 16) tissues. c Kaplan-Meier survival curves comparing osteosarcoma patients with low versus high ASB7 protein expression (low, n = 19; high, n = 19). The cutoff value for distinguishing the low- and high-expression groups was defined as the median IHC score obtained from the HALO system. The corresponding P value from the log-rank test is shown. d , e , h 143B cells stably expressing doxycycline-inducible ASB7 overexpression were treated with or without 0.5 μg/mL doxycycline for 24 h. After that, the cells were subjected to western blotting to verify the upregulation of ASB7 expression ( d ), immunofluorescence staining with Actin-Tracker ( e ), and migration and invasion assays ( n = 3 independent experiments) ( h ). f The percentage of cells with protrusions was quantified in 143B cells overexpressing ASB7. Cells exhibiting finger-like filopodia (marked by asterisks) or fan-/wave-like lamellipodia (indicated by arrowheads), as shown in e , were scored as positive. g GO analysis of genes upregulated in 143B cells overexpressing ASB7. i – l Luciferase-expressing 143B cells with a doxycycline-inducible ASB7 overexpression system were injected into mouse tibiae. After that, the mice were fed either a control diet or a diet containing 0.065% doxycycline. Lung metastases were assessed by IVIS imaging ( i ) and quantitative bioluminescence analysis ( j ). Lung tissue sections were subjected to H&E staining ( k ), and metastatic foci were quantified ( l ). n = 5 mice per group. m – q 143B cells expressing sgRNAs of either non-target control or ASB7 were subjected to western blotting to detect the expression of ASB7 ( m ), immunofluorescence staining with Actin-Tracker ( n ), a quantification of cells with finger-like or fan-/wave-like protrusions ( o ), migratory and invasive assays ( n = 3 independent experiments) ( p ), and an MTT assay ( n = 3 independent experiments) ( q ). r – u Mice implanted with 143B cells expressing both luciferase and sgRNAs targeting either non-target control or ASB7 were analyzed by bioluminescence imaging, IVIS images are shown ( r ), and the data are plotted ( s ). Lung sections from these mice were analyzed by H&E staining ( t ), and the quantification of metastatic foci ( u ) is plotted. n = 5 mice per group. Data are shown as the mean ± S.D. P values were derived from unpaired two-tailed Student’s t -test for the data in b , f , h , j , l , o , p , s , u and from two-way ANOVA followed by Tukey’s multiple comparisons test for the data in q .

Article Snippet: The human cell lines 143B and HEK293T were purchased from ATCC.

Techniques: Expressing, RNA Sequencing, Stable Transfection, Over Expression, Western Blot, Immunofluorescence, Staining, Migration, Luciferase, Injection, Control, Imaging, MTT Assay, Derivative Assay, Two Tailed Test

Journal: Cell Discovery

Article Title: ASB7 promotes osteosarcoma lung metastasis through ubiquitin-mediated degradation of ATF2

doi: 10.1038/s41421-026-00890-9

Figure Lengend Snippet: a The normalized protein levels of ASB7 and ATF2 across unbiased re-classified tumor subtypes were plotted using data from the UALCAN database , . Pearson’s correlation coefficient ( r ) was calculated to assess the relationship between ASB7 and ATF2 protein levels across tumor subtypes. b Representative images of immunohistochemical staining of ASB7 and ATF2 in osteosarcoma tissues. c A contingency table was constructed to display the stratification of 38 osteosarcoma cases based on high or low expression of ASB7 and ATF2, in which the dichotomization was determined by the median IHC score obtained from the HALO system. The Spearman correlation coefficient was used to quantify the association, while Pearson’s chi-square test was used to determine the corresponding χ 2 statistic and P value. d Kaplan-Meier survival analysis of osteosarcoma patients based on ATF2 protein expression level (low, n = 19; high, n = 19). The P value from the log-rank test is shown, with the high and low-expression groups dichotomized by the median IHC score obtained from the HALO system. e – j 143B cells expressing vector or ATF2 were subjected to western blotting ( e ), Actin-Tracker staining ( f ), quantification of cells exhibiting finger-like or fan-/wave-like protrusions ( g ), cell migration and invasion assays ( n = 3 independent experiments) ( i ), and MTT assays ( n = 3 independent experiments) ( j ). h GO analysis of genes upregulated in 143B cells in which ATF2 was knocked down by siRNAs. k , l In vivo fluorescence imaging and the corresponding quantitative analysis ( k ), H&E staining and the corresponding quantitative analysis ( l ) of mice bearing orthotopic 143B cells expressing luciferase together with either vector or ATF2 ( n = 5 mice per group). m – q In 143B cells expressing sgRNAs targeting either non-target control or ATF2, western blotting ( m ), Actin-Tracker staining ( n ), quantification of cells with finger-like or fan-/wave-like protrusions ( o ), cell migration and invasion assays ( n = 3 independent experiments) ( p ), and MTT assays ( n = 3 independent experiments) ( q ) were performed. r, s In mice bearing orthotopic 143B cells expressing luciferase together with sgRNAs targeting non-target control or ATF2, in vivo fluorescence imaging ( r ), H&E staining ( s ) of lung sections and the corresponding quantitative analyses were performed ( n = 5 mice per group). Data are shown as the mean ± S.D. P values were derived from unpaired two-tailed Student’s t -test for the data in g , i , k , l , o , p , r , s ; from two-way ANOVA followed by Sidak’s test for the data in j ; or from two-way ANOVA followed by Tukey’s test for the data in q .

Article Snippet: The human cell lines 143B and HEK293T were purchased from ATCC.

Techniques: Immunohistochemical staining, Staining, Construct, Expressing, Plasmid Preparation, Western Blot, Migration, In Vivo, Fluorescence, Imaging, Luciferase, Control, Derivative Assay, Two Tailed Test

Journal: Cell Discovery

Article Title: ASB7 promotes osteosarcoma lung metastasis through ubiquitin-mediated degradation of ATF2

doi: 10.1038/s41421-026-00890-9

Figure Lengend Snippet: a , b ATF2 protein levels were analyzed by western blotting in 143B cells with ASB7 overexpression ( a ) or knockout ( b ). c Western blotting analysis of ATF2 protein levels in 143B cells treated with MG132 (10 μM, 6 h), MLN4924 (1 μM, 12 h), or bafilomycin A1 (200 nM, 6 h). d – g Western blotting analysis of ATF2 protein levels at the indicated times following CHX (40 μg/mL) treatment in 143B cells with ASB7 overexpression ( d ) or knockout ( f ). Panels e and g show the corresponding quantitative data from these experiments. n = 3 independent experiments. h HEK293T cells were co-transfected with the indicated constructs for 24 h and treated with 10 μM MG132 for another 6 h. After that, the cells were lysed and subjected to immunoprecipitation using anti-Flag or anti-V5 beads. i HEK293T cells were co-transfected with the indicated constructs for 24 h and subsequently treated with MG132 (10 μM, 6 h). After that, the cells were subjected to a Ni-NTA pull-down assay to examine the exogenous ubiquitination of ATF2. j , k Endogenous ubiquitination of ATF2 was evaluated by Ni-NTA pull-down assay in 143B cells with ASB7 overexpression ( j ) or knockout ( k ) following transfection with His-Ub and treatment with 10 μM MG132 for 6 h. l Ubiquitination analysis of ATF2 mutants. HEK293T cells were co-transfected with the indicated constructs, treated with MG132 (10 μM, 6 h), and subjected to a Ni-NTA pull-down assay to assess ATF2 ubiquitination levels. Data are shown as the mean ± S.D. P values were derived from two-way ANOVA followed by Sidak’s multiple comparisons test, as shown in ( e ), or two-way ANOVA followed by Tukey’s multiple comparisons test, as shown in g .

Article Snippet: The human cell lines 143B and HEK293T were purchased from ATCC.

Techniques: Western Blot, Over Expression, Knock-Out, Transfection, Construct, Immunoprecipitation, Pull Down Assay, Ubiquitin Proteomics, Derivative Assay

Journal: Cell Discovery

Article Title: ASB7 promotes osteosarcoma lung metastasis through ubiquitin-mediated degradation of ATF2

doi: 10.1038/s41421-026-00890-9

Figure Lengend Snippet: a GO analysis of genes upregulated in both 143B cells with ASB7 overexpression and those with ATF2 knockdown by siRNAs. b – e 143B cells expressing doxycycline-induced ASB7 together with either vector or ATF2 were subjected to western blotting ( b ), Actin-Tracker staining ( c ), quantification of cells with finger-like or fan-/wave-like protrusions ( d ), and migration and invasion assays ( e ). n = 3 independent experiments. f – i In mice bearing orthotopic 143B-Luc cells expressing doxycycline-induced ASB7 together with either vector or ATF2, in vivo lung fluorescence images ( f ) and H&E staining of lung sections ( h ) are shown, along with the corresponding quantitative analyses in g , i , respectively. n = 5 mice per group. Data are shown as the mean ± S.D. Statistical significance in d , e , g , i was assessed by an unpaired two-tailed Student’s t -test.

Article Snippet: The human cell lines 143B and HEK293T were purchased from ATCC.

Techniques: Over Expression, Knockdown, Expressing, Plasmid Preparation, Western Blot, Staining, Migration, In Vivo, Fluorescence, Two Tailed Test

Journal: Cell Discovery

Article Title: ASB7 promotes osteosarcoma lung metastasis through ubiquitin-mediated degradation of ATF2

doi: 10.1038/s41421-026-00890-9

Figure Lengend Snippet: a The ATF2 peak enrichment scores and expression fold changes for migration-related genes upregulated upon ASB7 overexpression were plotted. The dashed red line indicates the ideal distribution of genes that are both strongly bound by ATF2 and upregulated upon ASB7 overexpression. b Pearson’s correlation coefficients ( r ) and the corresponding P values of ATF2 and ITGB2 expression across tumor types were plotted using TCGA transcriptomic data from the GEPIA database. c Schematic of the ITGB2 promoter region (top) and the subsequent ChIP-qPCR assay used to identify the site enriched in ATF2 in 143B cells. d – k ITGB2 protein levels were analyzed by western blotting in 143B cells with ASB7 overexpression ( d ), ASB7 knockout ( f ), ATF2 overexpression ( h ), and ATF2 knockout ( j ), respectively. ITGB2 mRNA levels were analyzed by qRT-PCR in 143B cells with ASB7 overexpression ( e ), ASB7 knockout ( g ), ATF2 overexpression ( i ), and ATF2 knockout ( k ), respectively. l – p 143B cells expressing vector or ITGB2 were analyzed using western blotting ( l ) and Actin-Tracker staining ( m ). The proportion of cells with finger-like or fan-/wave-like protrusion morphology was quantified ( n ). Additionally, cell migration and invasion assays ( o ) and MTT assays ( p ) were performed. q – u 143B cells expressing sgRNAs targeting either non-target control or ITGB2 were subjected to western blotting ( q ), Actin-Tracker staining ( r ), quantification of cells with finger-like or fan-/wave-like protrusions ( s ), cell migration and invasion assays ( t ), and MTT assays ( u ). The data are shown as the mean ± S.D. from three independent experiments. P values were derived from unpaired two-tailed Student’s t -test for the data in c , e , g , i , k , n , o , s , t and from two-way ANOVA followed by either Sidak’s or Tukey’s multiple comparisons tests for the data in p , u , respectively.

Article Snippet: The human cell lines 143B and HEK293T were purchased from ATCC.

Techniques: Expressing, Migration, Over Expression, ChIP-qPCR, Western Blot, Knock-Out, Quantitative RT-PCR, Plasmid Preparation, Staining, Control, Derivative Assay, Two Tailed Test

Journal: Cell Discovery

Article Title: ASB7 promotes osteosarcoma lung metastasis through ubiquitin-mediated degradation of ATF2

doi: 10.1038/s41421-026-00890-9

Figure Lengend Snippet: a HEK293T cells were co-transfected with V5-ATF2 and Flag-HDACs, and the interaction between ATF2 and HDACs was assessed by immunoprecipitation with anti-Flag beads. b qRT-PCR analysis of ITGB2 mRNA levels in 143B cells after knockdown of HDAC3, HDAC6, or HDAC10. c qRT-PCR analysis of ITGB2 mRNA levels in 143B cells with the indicated constructs. d Co-immunoprecipitation analysis of the endogenous interaction between HDAC6 and ATF2 in 143B cells. e , f 143B cells stably expressing vector or HDAC6 were analyzed by western blotting ( e ) and cell migration and invasion assays ( n = 3 independent experiments) ( f ). g , h 143B cells expressing shRNAs targeting either non-target control or HDAC6 were subjected to western blotting ( g ) and cell migration and invasion assays ( n = 3 independent experiments) ( h ). i Heatmaps showing the genome-wide distributions of ATF2 ChIP-seq signals (ENCODE) and HDAC6 CUT&Tag signals across gene bodies with ± 3.0 kb flanking regions. j Heatmaps showing the distributions of ATF2 ChIP-seq signals (ENCODE) and HDAC6 CUT&Tag signals centered on the TSSs of ATF2-bound promoters associated with migration-related genes. The signals are plotted ± 3.5 kb around the TSS. k , l ChIP-qPCR analysis of HDAC6 enrichment at the ITGB2 promoter in 143B cells with ATF2 overexpression ( k ) or knockout ( l ). m Model of the ASB7-ATF2/HDAC6-ITGB2 axis in osteosarcoma. ATF2 recruits HDAC6 to transcriptionally repress ITGB2 and inhibit cell movement, while ASB7-mediated ATF2 degradation relieves this repression, activating ITGB2 expression and inducing protrusion formation to drive metastasis. The data are shown as the mean ± S.D. from three independent experiments. P values were derived from unpaired two-tailed Student’s t -test, as shown in b , c , f , h , k , l .

Article Snippet: The human cell lines 143B and HEK293T were purchased from ATCC.

Techniques: Transfection, Immunoprecipitation, Quantitative RT-PCR, Knockdown, Construct, Stable Transfection, Expressing, Plasmid Preparation, Western Blot, Migration, Control, Genome Wide, ChIP-sequencing, ChIP-qPCR, Over Expression, Knock-Out, Derivative Assay, Two Tailed Test

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: Identification of active compounds and target prediction in YHD. (A) Venn diagram of the target of YHD and the target of osteosarcoma. (B – D) Gene Ontology (GO) enrichment analysis results. (E, F) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis results. (G) The component-target-pathway-disease network implicated in the mechanism of YHD in osteosarcoma treatment. The triangles represent osteosarcoma, the diamonds represent pathways, the circles represent key genes, and the squares represent the active ingredients of YHD. (H) Heatmap of molecular docking score. A binding energy heatmap with a bluer color indicates a more stable binding. (I) Molecular docking visualization between the active components of YHD and key targets.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Binding Assay

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD selectively inhibits osteosarcoma (OS) cells without affecting the viability or apoptosis of normal human cells. (A, B) CCK8 assay detected the effect of YHD on the viability of OS cells at 24 h and 48 h. (C, D) Colony formation assay detected the effect of YHD on the colony-forming ability of OS cells. (E – G) Flow cytometry was used to detect the effect of YHD on the cell cycle of OS cells. (H – J) Western blot analysis detected the effect of YHD on the levels of proliferation-related proteins in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Colony Assay, Flow Cytometry, Western Blot, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD can inhibit the migration and invasion of osteosarcoma (OS) cells, and promote their apoptosis. (A, B) Scratch healing assay showed that YHD inhibited the migration of OS cells. (C, D) Transwell assay showed that YHD inhibited the invasion of OS cells. (E – G) Western blot analysis detected the effect of YHD on the levels of proteins related to migration and invasion in OS cells. (H–K) Flow cytometry was used to detect the effect of YHD on apoptosis in OS cells. (L – N) Western blot analysis detected the effect of YHD on the levels of apoptosis-related proteins in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Migration, Transwell Assay, Western Blot, Flow Cytometry, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD induces osteosarcoma (OS) cell death by increasing ROS levels. (A) The effect of YHD on ROS levels in OS cells was detected using the DCFH probe method. (B) After N-acetylcysteine (NAC) treatment, the effect of YHD on ROS levels in OS cells was detected using the DCFH probe method. (C) After NAC treatment, CCK8 assay detected the effect of YHD on the viability of OS cells at 24 h and 48 h. (D, E) After NAC treatment, scratch healing assay showed that YHD inhibited the migration of OS cells. (F, G) After NAC treatment, Transwell assay showed that YHD inhibited the invasion of OS cells. (H, I) After NAC treatment, flow cytometry was used to detect the effect of YHD on the apoptosis of OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD can induce mitochondrial dysfunction in osteosarcoma (OS) cells. (A) Real-time quantitative PCR was used to measure mitochondrial DNA (mtDNA) levels. (B, C) Western blot analysis detected the effect of YHD on the expression levels of proteins related to mitochondrial biogenesis in OS cells. (D, E) JC-1 staining detected the effect of YHD on the mitochondrial membrane potential in OS cells. (F, G) MitoSOX staining detected the effect of YHD on mitochondrial ROS levels in OS cells. (H) Seahorse XFe24 analyzer measured the effect of YHD on the oxygen consumption rate (OCR) in OS cells. (I) The effect of YHD on ATP content in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, Membrane, Standard Deviation

Journal: Genes & Diseases

Article Title: Network pharmacology reveals that Yanghe Decoction inhibits osteosarcoma progression via ROS-induced mitochondrial dysfunction and enhances cisplatin sensitivity

doi: 10.1016/j.gendis.2025.101862

Figure Lengend Snippet: YHD exerts anti-tumor effects on osteosarcoma (OS) cells through the PI3K/AKT and p38 signaling pathways. (A) Principal component analysis revealed a clear distinction in gene expression profiles between the control and YHD groups. (B) Volcano plot identified 3495 differentially expressed genes in the YHD group. (C) Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. (D – G) Gene Set Enrichment Analysis (GSEA) of control and YHD groups. (H, I) Western blot analysis detected the effect of YHD on proteins related to the PI3K/AKT and MAPK pathways in OS cells. (J, K) After the addition of a PI3K activator and a P38 inhibitor, scratch healing assay showed that YHD inhibited the migration of OS cells. (L, M) After the addition of a PI3K activator and a P38 inhibitor, JC-1 staining detected the effect of YHD on the mitochondrial membrane potential in OS cells. Data were presented as mean ± standard deviation ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01 versus the blank group.

Article Snippet: Human osteosarcoma cell lines (HOS, 143B), human bone marrow stromal cells (HS-5), human proximal renal tubular epithelial cells (HK-2), and human normal liver cells (LO2) were all purchased from the American Type Culture Collection (ATCC).

Techniques: Protein-Protein interactions, Gene Expression, Control, Western Blot, Migration, Staining, Membrane, Standard Deviation