Journal: Investigative Ophthalmology & Visual Science

Article Title: SOX4 as a Key Oncogene Driving Tumor Invasion in Retinoblastoma

doi: 10.1167/iovs.66.5.24

Figure Lengend Snippet: SOX4 is associated with the malignant features of retinoblastoma. ( A , B ) Representative images of SOX4 immunohistochemistry ( A ) and immunofluorescence staining ( B ) in intraocular RB and extraocular RB patient samples. ( C , D ) SOX4 protein levels ( C ) and their quantitative analysis ( D ) in intraocular RB and extraocular RB. ( E – G ) Relative SOX4 gene expression ( E ), SOX4 protein levels ( F ), and representative immunofluorescence staining ( G ) in ARPE-19, WERI-RB1, and Y79 cell lines. Data are presented as mean ± SD. Statistical significance of panel ( D ) was determined using Student's t -test, while panel ( E ) used one-way ANOVA followed by Tukey's HSD post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: WERI-Rb1 and Y79 RB cell lines, along with the ARPE-19 human retinal pigment epithelial cell line (ATCC, Manassas, VA, USA), were used.

Techniques: Immunohistochemistry, Immunofluorescence, Staining, Gene Expression

Journal: Investigative Ophthalmology & Visual Science

Article Title: SOX4 as a Key Oncogene Driving Tumor Invasion in Retinoblastoma

doi: 10.1167/iovs.66.5.24

Figure Lengend Snippet: SOX4 promotes retinoblastoma proliferation in vitro. ( A ) Relative SOX4 mRNA expression in the Y79 cell line following siRNA transfection. ( B ) SOX4 protein expression levels in Y79 cells post-siRNA transfection. ( C ) optical density (OD) values measured at 450 nm in si-NC and si-SOX4 groups by CCK-8 assay. ( D ) EdU-positive cells and quantitative analysis ( E ) in si-NC and si-SOX4 groups. ( F , G ) Colony formation assay ( F ) and quantitative analysis ( G ) in si-NC and si-SOX4 groups. Data are presented as mean ± SD. Statistical significance of panel ( A ) was determined using one-way ANOVA followed by Tukey's HSD post hoc test. For panel ( C ), a two-way ANOVA followed by Šidák's post hoc test was used. In panels ( E ) and ( G ), Student's t-test was applied, with * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: WERI-Rb1 and Y79 RB cell lines, along with the ARPE-19 human retinal pigment epithelial cell line (ATCC, Manassas, VA, USA), were used.

Techniques: In Vitro, Expressing, Transfection, CCK-8 Assay, Colony Assay

Journal: Investigative Ophthalmology & Visual Science

Article Title: SOX4 as a Key Oncogene Driving Tumor Invasion in Retinoblastoma

doi: 10.1167/iovs.66.5.24

Figure Lengend Snippet: SOX4 involves EMT and migration of retinoblastoma. ( A ) Relative mRNA expression of the SNAI1 and SNAI2 genes, which involve EMT, in the si-NC and si-SOX4 groups. ( B ) Relative mRNA expression levels of the EMT biomarker genes CDH1 (encoding E-cadherin), CDH2 (encoding N-cadherin), and VIM (encoding vimentin). ( C , D ) Protein levels ( C ) and quantitative analysis ( D ) of N-cadherin, E-cadherin, and vimentin in si-NC and si-SOX4 groups. ( E ) Migrated Y79 cells stained with Hoechst 33342 in a transwell assay for si-NC and si-SOX4 groups. ( F ) Quantitative analysis of transwell assay for si-NC and si-SOX4 groups. Data are presented as mean ± SD. Statistical significance was determined using Student's t -test, with * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: WERI-Rb1 and Y79 RB cell lines, along with the ARPE-19 human retinal pigment epithelial cell line (ATCC, Manassas, VA, USA), were used.

Techniques: Migration, Expressing, Biomarker Discovery, Staining, Transwell Assay

Journal: Frontiers in Medicine

Article Title: Xanthatin induces apoptosis through ROS-mediated c-FLIP inhibition in human retinoblastoma cells

doi: 10.3389/fmed.2025.1554934

Figure Lengend Snippet: Effects of xanthatin on cell survival of retinoblastoma cell lines Y79 and WERI-RB-1. (A,B) Histograms showing cell survival (% of control) of Y79 (A) and WERI-RB-1 (B) cells treated with varying concentrations of xanthatin (0, 2.5, 5, 10, 20, 40 μM) for 24, 36, and 48 h. Cell survival decreases with increasing xanthatin concentration and treatment duration, with a more pronounced reduction at higher concentrations (20 and 40 μM) and longer time points (48 h). (C,D) Dose-response curves illustrating the cell survival (% of control) of Y79 (C) and WERI-RB-1 (D) cells treated with xanthatin at different concentrations (0 to 2.0 μM) over 24, 36, and 48 h. Cell survival decreases progressively with increasing xanthatin concentration, with a steeper decline at 48 h compared to 24 h and 36 h.

Article Snippet: The Y79 retinoblastoma cell line was obtained from ATCC, and the WERI-RB-1 cell line was obtained from Procell Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Control, Concentration Assay

Journal: Frontiers in Medicine

Article Title: Xanthatin induces apoptosis through ROS-mediated c-FLIP inhibition in human retinoblastoma cells

doi: 10.3389/fmed.2025.1554934

Figure Lengend Snippet: Xanthatin induces caspase-dependent apoptosis in a concentration-dependent manner in retinoblastoma cells. (A) Gene expression analysis from the GEO dataset (GSE208143) comparing the retinoblastoma (RB) group to the normal control (NC) group, showed a decrease in caspase-3 expression and an increase in caspase-8 and c-FLIP expression in the RB group. (B) Gene Set Enrichment Analysis (GSEA) from GSE208143, highlighting enrichment of the apoptosis gene set in the RB group, with a normalized enrichment score (NES) of 2.145 and a p -value <0.01. (C,D) Flow cytometry analysis of apoptosis in Y79 (C) and WERI-RB-1 (D) cell lines treated with xanthatin at concentrations of 0, 5, 10, and 20 μM for 48 h. (E) Quantification of apoptosis rates in Y79 and WERI-RB-1 cells treated with xanthatin (0, 5, 10, 20 μM) for 48 h, corresponding to the flow cytometry results in C,D . (F) Western blot analysis showing the expression levels of caspase-8, caspase-9, caspase-3, cleaved caspase-3, and cleaved PARP in Y79 and WERI-RB-1 cells treated with xanthatin (0, 5, 10, 20 μM).

Article Snippet: The Y79 retinoblastoma cell line was obtained from ATCC, and the WERI-RB-1 cell line was obtained from Procell Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Concentration Assay, Gene Expression, Control, Expressing, Flow Cytometry, Western Blot

Journal: Frontiers in Medicine

Article Title: Xanthatin induces apoptosis through ROS-mediated c-FLIP inhibition in human retinoblastoma cells

doi: 10.3389/fmed.2025.1554934

Figure Lengend Snippet: c-FLIP contributes to xanthatin-induced apoptosis. (A) Y79 and WERI-RB-1 cells were treated with 0, 5, 10, or 20 μM xanthatin for 24 h and then harvested to test the level of c-FLIP. (B) Y79 and WERI-RB-1 cells were transfected with pENTER or pENTER-cFLIP-FALG plasmids. After 48 h of transfection, the cells were exposed to 10 μM xanthatin for 24 h and then harvested to test the expression of cFLIP and apoptosis-related markers (caspase-8 caspase-9, caspase-3, and PARP). (C) Flow cytometry of Y79 cells treated with 10 μM xanthatin for 24 h, corresponds to B . (D) Flow cytometry of WERI-RB-1 cells treated with 10 μM xanthatin for 24 h, corresponds to B . (E,F) Data analysis was performed using FlowJo software and SPSS software. All data are presented as the mean ± SD.

Article Snippet: The Y79 retinoblastoma cell line was obtained from ATCC, and the WERI-RB-1 cell line was obtained from Procell Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Transfection, Expressing, Flow Cytometry, Software

Journal: Frontiers in Medicine

Article Title: Xanthatin induces apoptosis through ROS-mediated c-FLIP inhibition in human retinoblastoma cells

doi: 10.3389/fmed.2025.1554934

Figure Lengend Snippet: Xanthatin increases ROS production in retinoblastoma cells in a dose-dependent manner, and NAC weakens ROS production and survival inhibition induced by xanthatin. Xanthatin can increase ROS production in RB cells in a dose-dependent manner, and NAC can protect RB cells by reducing ROS production through antioxidation. (A) Y79 and WERI-RB-1 cells were treated with 0, 5, 10, or 20 μM xanthatin for 8 h and then harvested for flow cytometry to test the level of ROS. (B) Y79 and WERI-RB-1 cells were pretreated with NAC (5 mM) for 2 h before exposure to xanthatin for 8 h. Then, the intracellular ROS levels were measured by flow cytometry. (C,D) Flow cytometry of ROS levels in Y79 and WERI-RB1 cells pretreated with NAC (0, 2.5, 5, 10 μM) for 2 h, then treated with 10 μM xanthatin for 24 h.

Article Snippet: The Y79 retinoblastoma cell line was obtained from ATCC, and the WERI-RB-1 cell line was obtained from Procell Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Inhibition, Flow Cytometry

Journal: Frontiers in Medicine

Article Title: Xanthatin induces apoptosis through ROS-mediated c-FLIP inhibition in human retinoblastoma cells

doi: 10.3389/fmed.2025.1554934

Figure Lengend Snippet: NAC can attenuate xanthatin-induced apoptosis in retinoblastoma cells by reducing ROS production. Y79 and WERI-RB-1 cells were pretreated with NAC (5 mM) for 2 h before exposure to xanthatin for another 24 h. Then, cell apoptosis was measured by flow cytometry (A,B) . (C,D) Visualized the results of A,B . Western blot of apoptosis markers in Y79 and WERI-RB-1 cells pretreated with NAC (0 or 10 μM) for 2 h, then treated with xanthatin (0 or 10 μM) for 24 h. Xanthatin decreased c-FLIP(L) expression and increased cleavage of caspase-8, caspase-9, caspase-3, and PARP, while NAC attenuated these effects (E) .

Article Snippet: The Y79 retinoblastoma cell line was obtained from ATCC, and the WERI-RB-1 cell line was obtained from Procell Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Flow Cytometry, Western Blot, Expressing

Journal: Frontiers in Medicine

Article Title: Xanthatin induces apoptosis through ROS-mediated c-FLIP inhibition in human retinoblastoma cells

doi: 10.3389/fmed.2025.1554934

Figure Lengend Snippet: Xanthatin triggers endoplasmic reticulum stress in human retinoblastoma cells. Xanthatin triggers endoplasmic reticulum stress in human retinoblastoma cells. Y79 and WERI-RB-1 cells were treated with 0, 5, 10, and 20 μM xanthatin and incubated for 24 h. Following treatment, endoplasmic reticulum stress-related proteins were quantified by western blotting analysis.

Article Snippet: The Y79 retinoblastoma cell line was obtained from ATCC, and the WERI-RB-1 cell line was obtained from Procell Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Incubation, Western Blot

Journal: Frontiers in Medicine

Article Title: Xanthatin induces apoptosis through ROS-mediated c-FLIP inhibition in human retinoblastoma cells

doi: 10.3389/fmed.2025.1554934

Figure Lengend Snippet: Xanthatin inhibits human retinoblastoma cell tumorigenesis in vivo . (A) The xenograft tumors collected from the BALB/c nude male mice were randomly divided into a xanthatin-treaded group and a normal saline group after subcutaneously injected with Y79 cells for 15 days. (B) Tumor weight of mice treated with normal saline or xanthatin. Mean ± SD ( n = 5). *** p < 0.001 vs. normal saline. (C) Tumor volume of mice treated with normal saline or xanthatin. Mean ± SD ( n = 5). *** p < 0.001 vs. normal saline.

Article Snippet: The Y79 retinoblastoma cell line was obtained from ATCC, and the WERI-RB-1 cell line was obtained from Procell Biotechnology Co., Ltd. (Wuhan, China).

Techniques: In Vivo, Saline, Injection

Journal: Medicina

Article Title: Inhibition of Retinoblastoma Cell Growth by Boswellic Acid Through Activation of the Suppressing Nuclear Factor—κB Activation

doi: 10.3390/medicina61030480

Figure Lengend Snippet: Boswellic acid and Cisplatin cytotoxicity in Y79 cells. BA IC 50 and Cis IC 50 were determined after 24, 48 and 72 h of BA and Cis application. The figure shows cell viability (%) of Y79 cells treated with Cis and BA at different concentrations (0.5–100 μM) for 24 h, 48 h and 72 h. The bar graphs compare the effects of Cis (blue) and BA (orange) over time. Cell viability decreases in a dose- and time-dependent manner for both treatments. IC 50 values indicate that Cisplatin is slightly more effective than BA, with lower IC 50 values at 48 h (Cis: 2.18, BA: 2.06) and 72 h (Cis: 1.48, BA: 1.84). At higher concentrations (≥10 μM), both treatments significantly reduce cell viability.

Article Snippet: Human retinoblastoma cell line Y79 was obtained from American Type Culture Collection (ATCC ® HTB-18 TM , Manassas, VA, USA).

Techniques:

Journal: Medicina

Article Title: Inhibition of Retinoblastoma Cell Growth by Boswellic Acid Through Activation of the Suppressing Nuclear Factor—κB Activation

doi: 10.3390/medicina61030480

Figure Lengend Snippet: Effects of BA (IC 50 ), Cis (IC 50 ) and BA (IC 50 ) + Cis (IC 50 ) on inflammation in Y79 cells for 48 h. Inflammation markers TNF-α, IL1-β and IFN-γ were measured using ELISA kits. IL1-β and IFN-γ were determined as pg/mg and TNF- α as ng/mg. The bar graph represents the effects of Cis and BA on inflammatory cytokine levels (TNF-α, IL1-β, and IFN-γ) in Y79 cells. Cis treatment significantly decreased TNF-α, IL1-β and IFN-γ levels compared to the control group, with the greatest reduction observed in IL1-β. BA treatment did not significantly alter cytokine levels compared to the control group. BA + Cis treatment significantly reduced TNF-α and IL1-β levels ( p < 0.05), but the decrease in IFN-γ was not statistically significant ( p > 0.05).

Article Snippet: Human retinoblastoma cell line Y79 was obtained from American Type Culture Collection (ATCC ® HTB-18 TM , Manassas, VA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Medicina

Article Title: Inhibition of Retinoblastoma Cell Growth by Boswellic Acid Through Activation of the Suppressing Nuclear Factor—κB Activation

doi: 10.3390/medicina61030480

Figure Lengend Snippet: Relative fold increase values of p53 and Caspase-3 gene expressions in Y79 cell lines and decrease value of NF-κB expressions, IC 50 doses for 48 h after single and combined drug administration. The figure presents the mRNA expression levels of p53 , Caspase-3 and NF-κB under different treatment conditions: Control (Ctrl), Cis, BA and BA + Cis. The y -axis represents the fold change in mRNA expression, while the x -axis indicates the treatment groups. Statistical significance is indicated by p -values, with notable differences between groups. For p53 and Caspase-3 , mRNA expression is significantly increased in the Cis group compared to the Ctrl group ( p < 0.0001). The BA group also shows elevated expression, but the highest increase is observed in the BA + Cis combination group. For NF-κB , mRNA expression is also significantly upregulated in the Cis group compared to the Ctrl ( p < 0.0001). However, the BA group exhibits a lower NF-κB expression compared to Cis, and the BA + Cis group shows an intermediate level.

Article Snippet: Human retinoblastoma cell line Y79 was obtained from American Type Culture Collection (ATCC ® HTB-18 TM , Manassas, VA, USA).

Techniques: Expressing, Control