Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: Knockdown of XPO1 inhibited the invasion and migration abilities of ccRCC cells (A) The Venn diagram obtained by taking the intersection of the relevant gene sets of CCT3 and RB1 after screening. (B) Expression level of XPO1 in clear cell RCC from the TCGA database. (C) The immunohistochemical results of XPO1 in ccRCC and normal renal tissue in the HPA database. (D) A Kaplan-Meier curve illustrates OS in patients with ccRCC from low and high XPO1 expression groups. (E) The promoter methylation level of XPO1 in KIRC from the TCGA database. (F) Western blot analysis was performed to assess the protein expression of XPO1 in CCT3-depleted cells. (G) Using Spearman correlation analysis, the correlation between Gene CCT3 and XPO1 expression, and the correlation between XPO1 and RB1 expression. (H) Wound-healing assay indicates that XPO1 knockdown suppresses the viability of 786-O and 769-P cells. (I) Transwell assay indicates impaired abilities of migration and invasion of 769-P and 786-O cells. (Scale bars, 100 μm, all data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). Two-tailed Student’s t test for B and E. Student’s t test for F, H, and I.

Article Snippet: Selinexor, a selective XPO1 inhibitor, was purchased from MCE.

Techniques: Knockdown, Migration, Expressing, Immunohistochemical staining, Methylation, Western Blot, Wound Healing Assay, Transwell Assay, Two Tailed Test

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: The reduction of XPO1 induces cellular senescence in ccRCC cells (A) Western blot analysis was performed to assess the protein expression of CDK4, CyclinD, p-Rb (Ser807/811) in XPO1 knockdown cells. (B) The mRNA expression levels of senescence-associated genes, including interleukin 1 A, TNF, CDK2, CDK4, and CDK6 in XPO1-depleted cells were examined by qRT-PCR. (C) Changes of SA-β-gal activity in XPO1-knockdown cells. Data represent the percentage of cells staining positive for SA-β-gal ±SD. (D) Cell cycle analysis calculated the distribution of the cells in G1, S, and G2/M phases. (E) The proliferative abilities of 769-P and 786-O cells, which were treated with the XPO1 inhibitor Selinexor, were measured with an EdU staining assay. (Scale bars, 100 μm, all data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). Student’s t test.

Article Snippet: Selinexor, a selective XPO1 inhibitor, was purchased from MCE.

Techniques: Western Blot, Expressing, Knockdown, Quantitative RT-PCR, Activity Assay, Staining, Cell Cycle Assay

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: CCT3 regulates RB1 activity by influencing the stability of the XPO1 protein (A) The Co-IP was performed to analyze the endogenous interaction between CCT3 and XPO1 in ccRCC cells. protein pellets were analyzed by Western blot with anti-CCT3 and anti-XPO1 antibodies. (B) Pull-down of CCT3, XPO1, and CCT3 with GST-tagged was analyzed by Western Blot. (C) Western blot analysis of XPO1 in control or CCT3–knockdown cells treated with CHX for the times indicated. (D) Western blot analysis of XPO1 in control or CCT3–knockdown cells treated with BafA1. (E) Western blot analysis of XPO1 in control or CCT3–knockdown cells treated with MG132. (F) Visualization results of the CCT3 and XPO1 molecular docking. (All data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). One-way ANOVA for C, Student’s t test for D and E.

Article Snippet: Selinexor, a selective XPO1 inhibitor, was purchased from MCE.

Techniques: Activity Assay, Co-Immunoprecipitation Assay, Western Blot, Control, Knockdown

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: Rescue experiments demonstrate the effects of CCT3 overexpression and XPO1 knockdown on cell function (A) Western blot was used to detect the expression of RB1 and Cyclin D in nuclear and cytoplasmic fractions of cells. (B) Western blot analysis was performed to assess the protein expression of Cyclin D, p-Rb (Ser807/811) in the rescue assay. (C) Senescence β-Galactosidase Staining in rescue assay. (D) A healing assay was performed in the rescue assay. (E) Transwell assay was performed in the rescue assay. (Scale bars, 100 μm, all data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). One-way ANOVA for D, E.

Article Snippet: Selinexor, a selective XPO1 inhibitor, was purchased from MCE.

Techniques: Over Expression, Knockdown, Cell Function Assay, Western Blot, Expressing, Rescue Assay, Staining, Transwell Assay

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: Knockdown of CCT3 combined with a selective XPO1 inhibitor can promote cellular senescence to suppress tumor progression in vivo (A) Nude mice were used to establish a tumor xenograft model utilizing CCT3 knockdown cell lines and the selective XPO1 inhibitor, Selinexor. (B and C) The tumor volumes and tumor weight of each group. (D) Western blot analysis was performed to assess the protein expression of p53, Cyclin D, CDK4, and PCNA in tissue. (E and F) The immunohistochemistry and immunofluorescence assays of Ki67. (Scale bars, 100 μm. All data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). Repeated measures ANOVA for B and C. One-way ANOVA for D and E.

Article Snippet: Selinexor, a selective XPO1 inhibitor, was purchased from MCE.

Techniques: Knockdown, In Vivo, Western Blot, Expressing, Immunohistochemistry, Immunofluorescence

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: Knockdown of XPO1 inhibited the invasion and migration abilities of ccRCC cells (A) The Venn diagram obtained by taking the intersection of the relevant gene sets of CCT3 and RB1 after screening. (B) Expression level of XPO1 in clear cell RCC from the TCGA database. (C) The immunohistochemical results of XPO1 in ccRCC and normal renal tissue in the HPA database. (D) A Kaplan-Meier curve illustrates OS in patients with ccRCC from low and high XPO1 expression groups. (E) The promoter methylation level of XPO1 in KIRC from the TCGA database. (F) Western blot analysis was performed to assess the protein expression of XPO1 in CCT3-depleted cells. (G) Using Spearman correlation analysis, the correlation between Gene CCT3 and XPO1 expression, and the correlation between XPO1 and RB1 expression. (H) Wound-healing assay indicates that XPO1 knockdown suppresses the viability of 786-O and 769-P cells. (I) Transwell assay indicates impaired abilities of migration and invasion of 769-P and 786-O cells. (Scale bars, 100 μm, all data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). Two-tailed Student’s t test for B and E. Student’s t test for F, H, and I.

Article Snippet: The XPO1 inhibitor Selinexor, CHX, BafA1, and MG132 were all purchased from MCE, American.

Techniques: Knockdown, Migration, Expressing, Immunohistochemical staining, Methylation, Western Blot, Wound Healing Assay, Transwell Assay, Two Tailed Test

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: The reduction of XPO1 induces cellular senescence in ccRCC cells (A) Western blot analysis was performed to assess the protein expression of CDK4, CyclinD, p-Rb (Ser807/811) in XPO1 knockdown cells. (B) The mRNA expression levels of senescence-associated genes, including interleukin 1 A, TNF, CDK2, CDK4, and CDK6 in XPO1-depleted cells were examined by qRT-PCR. (C) Changes of SA-β-gal activity in XPO1-knockdown cells. Data represent the percentage of cells staining positive for SA-β-gal ±SD. (D) Cell cycle analysis calculated the distribution of the cells in G1, S, and G2/M phases. (E) The proliferative abilities of 769-P and 786-O cells, which were treated with the XPO1 inhibitor Selinexor, were measured with an EdU staining assay. (Scale bars, 100 μm, all data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). Student’s t test.

Article Snippet: The XPO1 inhibitor Selinexor, CHX, BafA1, and MG132 were all purchased from MCE, American.

Techniques: Western Blot, Expressing, Knockdown, Quantitative RT-PCR, Activity Assay, Staining, Cell Cycle Assay

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: CCT3 regulates RB1 activity by influencing the stability of the XPO1 protein (A) The Co-IP was performed to analyze the endogenous interaction between CCT3 and XPO1 in ccRCC cells. protein pellets were analyzed by Western blot with anti-CCT3 and anti-XPO1 antibodies. (B) Pull-down of CCT3, XPO1, and CCT3 with GST-tagged was analyzed by Western Blot. (C) Western blot analysis of XPO1 in control or CCT3–knockdown cells treated with CHX for the times indicated. (D) Western blot analysis of XPO1 in control or CCT3–knockdown cells treated with BafA1. (E) Western blot analysis of XPO1 in control or CCT3–knockdown cells treated with MG132. (F) Visualization results of the CCT3 and XPO1 molecular docking. (All data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). One-way ANOVA for C, Student’s t test for D and E.

Article Snippet: The XPO1 inhibitor Selinexor, CHX, BafA1, and MG132 were all purchased from MCE, American.

Techniques: Activity Assay, Co-Immunoprecipitation Assay, Western Blot, Control, Knockdown

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: Rescue experiments demonstrate the effects of CCT3 overexpression and XPO1 knockdown on cell function (A) Western blot was used to detect the expression of RB1 and Cyclin D in nuclear and cytoplasmic fractions of cells. (B) Western blot analysis was performed to assess the protein expression of Cyclin D, p-Rb (Ser807/811) in the rescue assay. (C) Senescence β-Galactosidase Staining in rescue assay. (D) A healing assay was performed in the rescue assay. (E) Transwell assay was performed in the rescue assay. (Scale bars, 100 μm, all data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). One-way ANOVA for D, E.

Article Snippet: The XPO1 inhibitor Selinexor, CHX, BafA1, and MG132 were all purchased from MCE, American.

Techniques: Over Expression, Knockdown, Cell Function Assay, Western Blot, Expressing, Rescue Assay, Staining, Transwell Assay

Journal: iScience

Article Title: CCT3-mediated regulation of XPO1/RB1 axis stability promotes cellular senescence and tumor progression in clear cell renal carcinoma

doi: 10.1016/j.isci.2026.114840

Figure Lengend Snippet: Knockdown of CCT3 combined with a selective XPO1 inhibitor can promote cellular senescence to suppress tumor progression in vivo (A) Nude mice were used to establish a tumor xenograft model utilizing CCT3 knockdown cell lines and the selective XPO1 inhibitor, Selinexor. (B and C) The tumor volumes and tumor weight of each group. (D) Western blot analysis was performed to assess the protein expression of p53, Cyclin D, CDK4, and PCNA in tissue. (E and F) The immunohistochemistry and immunofluorescence assays of Ki67. (Scale bars, 100 μm. All data were shown as the mean ± SD ( n = 3 per group), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001). Repeated measures ANOVA for B and C. One-way ANOVA for D and E.

Article Snippet: The XPO1 inhibitor Selinexor, CHX, BafA1, and MG132 were all purchased from MCE, American.

Techniques: Knockdown, In Vivo, Western Blot, Expressing, Immunohistochemistry, Immunofluorescence

Journal: Frontiers in Oncology

Article Title: Selinexor enhances the sensitivity of hepatocellular carcinoma cells to sorafenib by regulating the BAX/Bcl-2/PUMA apoptotic pathway and the XPO1/p27 cell cycle pathway

doi: 10.3389/fonc.2026.1762822

Figure Lengend Snippet: Inhibitory effect of combined selinexor and sorafenib on xenograft tumor growth in nude mice. (A) Photographs of final tumor volumes from different treatment groups. (B) Quantitative bar graph of final tumor volumes from different treatment groups. (C) Photographs of the final tumor weights from different treatment groups. (D) Quantitative bar graph of the final tumor weights from different treatment groups. (mean ± SD, n=5; Sel-10 = selinexor monotherapy (10 mg/kg), Sor-10 = sorafenib monotherapy (10 mg/kg), Sel-10 + Sor-10 = selinexor + sorafenib combination (10 mg/kg + 10 mg/kg); *P < 0.05, **P <0.01, ***P <0.001. Statistical analysis was performed using one-way ANOVA followed by LSD- t test). (Refer to Table 1 for detailed measured tumor volume data at key time points during the entire treatment period).

Article Snippet: Selinexor was purchased from Selleck Chemicals, and sorafenib was obtained from MedChemExpress (MCE).

Techniques:

Journal: Frontiers in Oncology

Article Title: Selinexor enhances the sensitivity of hepatocellular carcinoma cells to sorafenib by regulating the BAX/Bcl-2/PUMA apoptotic pathway and the XPO1/p27 cell cycle pathway

doi: 10.3389/fonc.2026.1762822

Figure Lengend Snippet: Immunohistochemical detection results of related proteins (BAX, Bcl-2, p27, and XPO1) in xenograft tumor tissues of nude mice from different treatment groups. (A) Representative immunohistochemical images of BAX, Bcl-2, p27, and XPO1 expression (magnification=40×; scale bar=50 μm). (B) Quantitative analysis of BAX expression among treatment groups. (C) Quantitative analysis of Bcl-2 expression among treatment groups. (D) Quantitative analysis of p27 expression among treatment groups. (E) Quantitative analysis of XPO1 expression among treatment groups. (Quantitative analysis of immunohistochemical staining intensity: mean ± SD, n=5 fields per section; Sel-10 = selinexor monotherapy (10 mg/kg), Sor-10 = sorafenib monotherapy (10 mg/kg), Sel-10 + Sor-10 = selinexor + sorafenib combination (10 mg/kg + 10 mg/kg); *P < 0.05, **P <0.01, ***P <0.001. Statistical analysis was performed using one-way ANOVA followed by LSD- t test). Coordinated modulation of BAX and Bcl-2 in tumor tissues is consistent with in vitro PUMA upregulation, supporting the functional activation of the BAX/Bcl-2/PUMA apoptotic pathway in vivo (refer to Section 3.1 and Discussion for details).

Article Snippet: Selinexor was purchased from Selleck Chemicals, and sorafenib was obtained from MedChemExpress (MCE).

Techniques: Immunohistochemical staining, Expressing, Staining, In Vitro, Functional Assay, Activation Assay, In Vivo

Journal: Frontiers in Oncology

Article Title: Selinexor enhances the sensitivity of hepatocellular carcinoma cells to sorafenib by regulating the BAX/Bcl-2/PUMA apoptotic pathway and the XPO1/p27 cell cycle pathway

doi: 10.3389/fonc.2026.1762822

Figure Lengend Snippet: Inhibitory effects of selinexor and sorafenib monotherapy on HCC cell proliferation (CCK-8 assay) (n=3). (A) Inhibitory effect of selinexor at different concentrations on Huh7 cell proliferation (24h, 48h, 72h). (B) Inhibitory effect of sorafenib at different concentrations on Huh7 cell proliferation (24h, 48h, 72h). (C) Inhibitory effect of selinexor at different concentrations on SK-HEP-1 cell proliferation (24h, 48h, 72h). (D) Inhibitory effect of sorafenib at different concentrations on SK-HEP-1 cell proliferation (24h, 48h, 72h). (E) Inhibitory effect of selinexor at different concentrations on HepG2 cell proliferation (24h, 48h, 72h). (F) Inhibitory effect of sorafenib at different concentrations on HepG2 cell proliferation (24h, 48h, 72h). (mean ± SD, n=3 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001; statistical significance was determined by two-way ANOVA followed by Bonferroni post-hoc test). (The IC 50 values of selinexor and sorafenib against Huh7, SK-HEP-1, and HepG2 cells at 72 h are provided in the main text (Section 3.2)).

Article Snippet: Selinexor was purchased from Selleck Chemicals, and sorafenib was obtained from MedChemExpress (MCE).

Techniques: CCK-8 Assay

Journal: Frontiers in Oncology

Article Title: Selinexor enhances the sensitivity of hepatocellular carcinoma cells to sorafenib by regulating the BAX/Bcl-2/PUMA apoptotic pathway and the XPO1/p27 cell cycle pathway

doi: 10.3389/fonc.2026.1762822

Figure Lengend Snippet: Inhibitory effects of combined selinexor and sorafenib on HCC cell proliferation (CCK-8 assay) (n=3). (A) Effect of combined selinexor and sorafenib on Huh7 cell proliferation (48h). (B) Effect of combined selinexor and sorafenib on Huh7 cell proliferation (72h). (C) Effect of combined selinexor and sorafenib on SK-HEP-1 cell proliferation (48h). (D) Effect of combined selinexor and sorafenib on SK-HEP-1 cell proliferation (72h). (E) Effect of combined selinexor and sorafenib on HepG2 cell proliferation (48h). (F) Effect of combined selinexor and sorafenib on HepG2 cell proliferation (72h). (mean ± SD, n=3 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001. Statistical analysis was performed using one-way ANOVA followed by LSD- t test).

Article Snippet: Selinexor was purchased from Selleck Chemicals, and sorafenib was obtained from MedChemExpress (MCE).

Techniques: CCK-8 Assay

Journal: Frontiers in Oncology

Article Title: Selinexor enhances the sensitivity of hepatocellular carcinoma cells to sorafenib by regulating the BAX/Bcl-2/PUMA apoptotic pathway and the XPO1/p27 cell cycle pathway

doi: 10.3389/fonc.2026.1762822

Figure Lengend Snippet: Regulatory effects of combined selinexor and sorafenib on cell cycle distribution of HCC cells (n=3). (A) Cell cycle distribution of Huh7 cells in the blank control group. (B) Cell cycle distribution of Huh7 cells in the sorafenib group. (C) Cell cycle distribution of Huh7 cells in the selinexor group. (D) Cell cycle distribution of Huh7 cells in the selinexor + sorafenib group. (E) Quantitative analysis of cell cycle distribution in Huh7 cells. (F) Cell cycle distribution of HepG2 cells in the blank control group. (G) Cell cycle distribution of HepG2 cells in the sorafenib group. (H) Cell cycle distribution of HepG2 cells in the selinexor group. (I) Cell cycle distribution of HepG2 cells in the selinexor + sorafenib group. (J) Quantitative analysis of cell cycle distribution in HepG2 cells. (Quantitative analysis of cell cycle phase proportions: mean ± SD, n=3 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001. Statistical analysis was performed using one-way ANOVA followed by LSD- t test).

Article Snippet: Selinexor was purchased from Selleck Chemicals, and sorafenib was obtained from MedChemExpress (MCE).

Techniques: Control

Journal: Frontiers in Oncology

Article Title: Selinexor enhances the sensitivity of hepatocellular carcinoma cells to sorafenib by regulating the BAX/Bcl-2/PUMA apoptotic pathway and the XPO1/p27 cell cycle pathway

doi: 10.3389/fonc.2026.1762822

Figure Lengend Snippet: Apoptosis-inducing effects of combined selinexor and sorafenib on HCC cells (n=3). (A) Apoptotic rate of Huh7 cells in the blank control group. (B) Apoptotic rate of Huh7 cells in the sorafenib group. (C) Apoptotic rate of Huh7 cells in the selinexor group. (D) Apoptotic rate of Huh7 cells in the selinexor + sorafenib group. (E) Quantitative analysis of apoptotic rate in Huh7 cells. (F) Apoptotic rate of HepG2 cells in the blank control group. (G) Apoptotic rate of HepG2 cells in the sorafenib group. (H) Apoptotic rate of HepG2 cells in the selinexor group. (I) Apoptotic rate of HepG2 cells in the selinexor + sorafenib group. (J) Quantitative analysis of apoptotic rate in HepG2 cells. (Quantitative analysis of total apoptotic rates (early + late apoptosis): mean ± SD, n=3 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001. Statistical analysis was performed using one-way ANOVA followed by LSD- t test).

Article Snippet: Selinexor was purchased from Selleck Chemicals, and sorafenib was obtained from MedChemExpress (MCE).

Techniques: Control

Journal: Frontiers in Oncology

Article Title: Selinexor enhances the sensitivity of hepatocellular carcinoma cells to sorafenib by regulating the BAX/Bcl-2/PUMA apoptotic pathway and the XPO1/p27 cell cycle pathway

doi: 10.3389/fonc.2026.1762822

Figure Lengend Snippet: Effects of combined selinexor and sorafenib on the expression of apoptosis- and cell cycle-related proteins (BAX, Bcl-2, p27, XPO1, PUMA) in Huh7 cells (n=3). (A) The protein expressions of BAX, Bcl-2, p27, XPO1, PUMA by Western Blot. (B) BAX protein data analysis. (C) Bcl-2 protein data analysis. (D) p27 protein data analysis. (E) XPO1 protein data analysis. (F) PUMA protein data analysis. (Quantitative analysis of protein expression levels (normalized to GAPDH): mean ± SD, n=3 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001. Statistical analysis was performed using one-way ANOVA followed by LSD- t test).

Article Snippet: Selinexor was purchased from Selleck Chemicals, and sorafenib was obtained from MedChemExpress (MCE).

Techniques: Expressing, Western Blot