Journal: Experimental and Therapeutic Medicine

Article Title: CDX2-UPK1B-PIK3IP1-PI3K/AKT signaling axis regulates gastric cancer cell invasion and migration and influences patient prognosis

doi: 10.3892/etm.2026.13179

Figure Lengend Snippet: UPK1B drives GC cell invasion and migration in a PI3K/AKT-dependent manner. (A) Gene set enrichment analysis indicated that genes upregulated in the UPK1B-high group were enriched in the PI3K/AKT pathway. (B) Protein levels of UPK1B in GC cell lines. (C) Knockdown of UPK1B reduced PI3K/AKT activation in MKN45 cells. Silencing UPK1B suppressed the (D) migration/invasion capacity and (E) wound closure rate of MKN45 cells. (F) Overexpression of UPK1B enhanced PI3K/AKT pathway activation in AGS cells, which was attenuated by the PI3K inhibitor LY294002. Inhibition of PI3K/AKT signaling reversed UPK1B-induced (G) migration/invasion capacity and (H) wound closure rate of AGS cells. UPK1B, uroplakin 1B; GC, gastric cancer; p-, phosphorylated; sh, short hairpin RNA; NC, negative control; OE, overexpression.

Article Snippet: Cells were also transfected with small interfering RNAs (siRNAs) targeting CDX2 or PIK3IP1 , with a universal non-targeting scrambled siRNA (si-NC) as the negative control , obtained from GeneChem, Inc. For UPK1B and CDX2 overexpression, the p-TSB-CMV-UPK1B and p-TSB-CMV-CDX2 expression vectors [Shanghai Genomeditech Co., Ltd.] and the corresponding empty p-TSB-CMV vector (negative control) were used.

Techniques: Migration, Knockdown, Activation Assay, Over Expression, Inhibition, shRNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: CDX2-UPK1B-PIK3IP1-PI3K/AKT signaling axis regulates gastric cancer cell invasion and migration and influences patient prognosis

doi: 10.3892/etm.2026.13179

Figure Lengend Snippet: CDX2 acts as a transcriptional repressor of UPK1B and its high expression is associated with favorable prognosis of patients with GC. (A) Venn diagram showing overlapping predicted transcriptional regulators of UPK1B from ChEA and ChEA3 databases. (B) Knockdown of CDX2 in AGS cells resulted in increased UPK1B (C) mRNA and (D) protein expression. (E) Overexpression of CDX2 in MKN45 cells reduced UPK1B protein levels. Data from (F) The Cancer Genome Atlas Stomach Adenocarcinoma cohort and (G) the Kaplan-Meier plotter database indicated that high CDX2 expression was associated with improved prognosis of patients with GC. UPK1B, uroplakin 1B; GC, gastric cancer; si, small interfering RNA; NC, negative control; OE, overexpression; HR, hazard ratio; CDX2, caudal-related homeobox transcription factor 2; ChEA, ChIP-X Enrichment Analysis.

Article Snippet: Cells were also transfected with small interfering RNAs (siRNAs) targeting CDX2 or PIK3IP1 , with a universal non-targeting scrambled siRNA (si-NC) as the negative control , obtained from GeneChem, Inc. For UPK1B and CDX2 overexpression, the p-TSB-CMV-UPK1B and p-TSB-CMV-CDX2 expression vectors [Shanghai Genomeditech Co., Ltd.] and the corresponding empty p-TSB-CMV vector (negative control) were used.

Techniques: Expressing, Knockdown, Over Expression, Small Interfering RNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: CDX2-UPK1B-PIK3IP1-PI3K/AKT signaling axis regulates gastric cancer cell invasion and migration and influences patient prognosis

doi: 10.3892/etm.2026.13179

Figure Lengend Snippet: UPK1B activates PI3K/AKT signaling by antagonizing the inhibitory regulator PIK3IP1 in gastric cancer cells. (A) Venn diagram showing that PIK3IP1 was identified as a putative UPK1B-interacting partner based on BioGRID and HIPPIE protein-protein interaction databases. (B) UPK1B and PIK3IP1 co-localized in the cytoplasm and plasma membrane of MKN45 cells. (C) Interaction between UPK1B and PIK3IP1 in MKN45 cells. (D) Knockdown of PIK3IP1 in MKN45 cells. (E) Silencing PIK3IP1 in UPK1B-knockdown MKN45 cells restored PI3K/AKT pathway activation. Knockdown of PIK3IP1 reversed the decrease in (F) migration/invasion and (G) wound-healing capacity in UPK1B-silenced MKN45 cells. UPK1B, uroplakin 1B; p-, phosphorylated; si, small interfering RNA; sh, short hairpin RNA; NC, negative control; PIK3IP1, PI3K inhibitor interacting protein 1; HIPPIE, Human Integrated Protein-Protein Interaction Reference; IP, immunoprecipitation.

Article Snippet: Cells were also transfected with small interfering RNAs (siRNAs) targeting CDX2 or PIK3IP1 , with a universal non-targeting scrambled siRNA (si-NC) as the negative control , obtained from GeneChem, Inc. For UPK1B and CDX2 overexpression, the p-TSB-CMV-UPK1B and p-TSB-CMV-CDX2 expression vectors [Shanghai Genomeditech Co., Ltd.] and the corresponding empty p-TSB-CMV vector (negative control) were used.

Techniques: Clinical Proteomics, Membrane, Knockdown, Activation Assay, Migration, Small Interfering RNA, shRNA, Negative Control, Immunoprecipitation

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