Journal: Cancer Biology & Therapy

Article Title: Centromere protein I promotes hepatocellular carcinoma progression by activating PI3K/AKT/mTOR-CDK2 cascade

doi: 10.1080/15384047.2026.2667596

Figure Lengend Snippet: Proliferation, survival, and invasion phenotypes were reshaped by gain- and loss-of-function perturbation. (A) WB analysis of CENPI protein in HCC cell lines (HCCLM3, Hep3B, and HepG2); HepG2 showed the highest CENPI expression ( n = 3). (B) WB analysis of CENPI protein in si-CENPI-transfected HepG2 cells; si-CENPI decreased CENPI levels vs si-NC, *** p < 0.001 ( n = 3). (C) WB analysis of CENPI protein in oe-CENPI-transfected HepG2 cells; oe-CENPI increased CENPI levels vs oe-NC, *** p < 0.001 ( n = 3). (D) CCK-8 assay for HepG2 proliferation; si-CENPI decreased absorbance (weaker proliferation) vs si-NC, while oe-CENPI increased it (stronger proliferation) vs oe-NC, * p < 0.05 ( n = 3). (E) Wound-healing assay for HepG2 migration; healing status recorded at 0/24 h. si-CENPI decelerated wound closure vs si-NC, while oe-CENPI accelerated it vs oe-NC; scale bar = 100 μm, ** p < 0.01 ( n = 3). (F) Flow cytometry for HepG2 apoptosis; apoptotic rates quantified. si-CENPI increased apoptotic rate vs si-NC, while oe-CENPI decreased it vs oe-NC, ** p < 0.01 ( n = 3). (G) Transwell assay for HepG2 invasion in Matrigel-coated chambers; invaded cells stained and counted. si-CENPI reduced invaded cells vs si-NC, while oe-CENPI increased them vs oe-NC; scale bar = 275 μm, *** p < 0.001 ( n = 3). (H) Flow cytometry for HepG2 cell cycle; si-CENPI increased the proportion of cells in G1 phase and decreased that in S phase vs si-NC, ** p < 0.01, while oe-CENPI showed the opposite effect vs oe-NC, * p < 0.05 ( n = 3). Data are presented as mean ± SD of three independent experiments; error bars represent SD.

Article Snippet: CENPI-targeting siRNAs and a nontargeting control siRNA (si-NC) were synthesized by Sangon Biotech (Shanghai, China).

Techniques: Expressing, Transfection, CCK-8 Assay, Wound Healing Assay, Migration, Flow Cytometry, Transwell Assay, Staining

Journal: Cancer Biology & Therapy

Article Title: Centromere protein I promotes hepatocellular carcinoma progression by activating PI3K/AKT/mTOR-CDK2 cascade

doi: 10.1080/15384047.2026.2667596

Figure Lengend Snippet: Cell cycle and mTORC1 programs emerged as dominant signatures accompanied by CDK2 elevation and EMT marker remodeling. (A) KEGG pathway enrichment analysis (top ten significantly enriched pathways); the cell cycle pathway was most significant, with significance represented by −log10 ( p value), based on CENPI-related differentially expressed genes. (B) GSEA analysis of CENPI-related genes; enriched pathways include cell cycle-related (E2F_TARGETS) and mechanism-related (mTORC1_SIGNALING), significance represented by normalized enrichment score (NES). (C) WB analysis of cell cycle-related proteins (CDK2, Cyclin D1) and CENPI in si-CENPI and oe-CENPI-transfected HCC cells; CDK2 showed the most obvious change, ** p < 0.01 vs si-NC and * p < 0.05 vs oe-NC ( n = 3). (D) WB detection of PI3K/AKT/mTOR pathway proteins (PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR), EMT proteins (N-cadherin, E-cadherin, Vimentin), and CENPI in si-CENPI and oe-CENPI-transfected HCC cells, * p < 0.05 and ** p < 0.01 ( n = 3). Quantitative data are presented as mean ± SD of three independent experiments; error bars represent SD.

Article Snippet: CENPI-targeting siRNAs and a nontargeting control siRNA (si-NC) were synthesized by Sangon Biotech (Shanghai, China).

Techniques: Marker, Transfection

Journal: Cancer Biology & Therapy

Article Title: Centromere protein I promotes hepatocellular carcinoma progression by activating PI3K/AKT/mTOR-CDK2 cascade

doi: 10.1080/15384047.2026.2667596

Figure Lengend Snippet: Orthotopic growth suppression was accompanied by pathway deactivation and reversal of EMT marker directionality. (A and B) Quantification of tumor weight in orthotopic xenograft models; shRNA-mediated CENPI silencing reduced tumor weight compared with models; scale bar = 1 cm, (C) Assessment of body weight changes; CENPI silencing attenuated cachexia-driven body weight loss, *** p < 0.001 vs control and ### p < 0.001 vs model ( n = 6). (D) Measurement of final tumor mass; CENPI silencing decreased tumor mass vs models without overt hepatotoxicity, ** p < 0.01 vs model ( n = 6). (E) WB analysis of EMT markers (E-cadherin, N-cadherin, and Vimentin) and PI3K/AKT/mTOR-CDK2 pathway proteins in resected tumor tissues. CENPI silencing induced a mesenchymal-to-epithelial reverting signature, with E-cadherin increased and N-cadherin/Vimentin suppressed. Concomitantly, p-PI3K, p-AKT, p-mTOR, and total CDK2 levels were diminished, indicating inactivation of the PI3K/AKT/mTOR-CDK2 relay, * p < 0.05 vs control, # p < 0.05, and ## p < 0.01 vs model ( n = 3). Data are presented as mean ± SD; error bars represent SD.

Article Snippet: CENPI-targeting siRNAs and a nontargeting control siRNA (si-NC) were synthesized by Sangon Biotech (Shanghai, China).

Techniques: Marker, shRNA, Control

Journal: Chinese Medicine

Article Title: Rhapontigenin alleviates cellular senescence and physiological aging by upregulating sirt1 and promoting autophagy

doi: 10.1186/s13020-025-01319-3

Figure Lengend Snippet: Rhap attenuates cellular senescence in part by upregulating Sirt1 and promoting autophagy. A – C NIH3T3 cells were cultured without or with 400 μM H 2 O 2 for 3 h. The next day, the medium was replaced with medium supplemented with 5, 10, 15, or 20 μM Rhap, and the cells were incubated for 48 h. The protein expression of p-Ampk, Ampk, and Sirt1 was measured by Western blotting, and the levels of p-Ampk/Ampk and Sirt1 were quantified. β-actin was used as the reference protein. D – F The protein expression of LC3B and p62 was measured by Western blotting, and the levels of LC3BII/LC3BI and p62 were quantified. β-actin was used as the reference protein. G Autophagosomes and autolysosomes (black arrows) in cells were observed by transmission electron microscopy. H The numbers of autophagic vacuoles (AVs), including autophagosomes and autolysosomes, were counted. I NIH3T3 cells were transfected with siRNA negative control (si-NC) or an siRNA against sirt1 (si-Sirt1). The protein expression of Sirt1 was measured by Western blotting. The representative results from three repetitions are shown in the figures, and the relative quantities are shown in the scatter bar chart. J Western blotting analysis of Sirt1, p62, LC3B, p21, and p16 protein expression in NIH3T3 cells transfected with si-NC or si-Sirt1 followed by treatment with or without H2O2 and Rhap at the concentrations shown in the figure. K Bar graph of the relative quantification of the gray values of the protein bands shown in Figure ( J ). The data are presented as the means ± SDs. Statistical analysis was performed with one-way ANOVA with Tukey's multiple comparisons or Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, ns, not significant

Article Snippet: The mouse negative control siRNA (si-NC) and siRNA targeting Sirt1 (si-Sirt1) were purchased from GeneChem (Shanghai, China).

Techniques: Cell Culture, Incubation, Expressing, Western Blot, Transmission Assay, Electron Microscopy, Transfection, Negative Control, Quantitative Proteomics

Journal: Journal of Orthopaedic Translation

Article Title: Pathomechanism of LYPD6 enhancing abnormal hyperostosis via Wnt/β-catenin signaling pathway in diffuse idiopathic skeletal hyperostosis

doi: 10.1016/j.jot.2025.10.003

Figure Lengend Snippet: (A, B) Protein and mRNA expression of β-Catenin in DISH-osteoblast lines, validated by WB and qRT-PCR. (C) LYPD6 siRNA knockdown in MC3T3 osteoblasts resulting in decreased β-Catenin protein expression. (D) Confocal immunofluorescence showing increased nuclear localization of β-Catenin in MC3T3-E1 cells after LYPD6 overexpression. (E) WB analysis showing downregulation of phosphorylated LRP6 following LYPD6 knockdown in MC3T3-E1 cells. (F) WB demonstrating upregulation of phosphorylated LRP6 after LYPD6 overexpression in MC3T3-E1 cells. (G) Co-immunoprecipitation analysis evaluating the effect of LRP6 and LYPD6 on osteoblast mineralization. (H) 3D structures of LYPD6 and LRP6 proteins, along with a prediction of the binding pattern between LYPD6 and LRP6. (∗P < 0.01, ∗∗P < 0.001, ∗∗∗P < 0.0001).

Article Snippet: LYPD6-specific siRNAs (GGAAAUAUCUGUAAUCUACCATT for human and ACUCAGAGCACGAAGGCUATT for mouse) and negative control siRNA (Si-NC) (KeyGEN, Nanjing, China) were used to knockdown LYPD6.

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Immunofluorescence, Over Expression, Immunoprecipitation, Binding Assay