Journal: Cellular & Molecular Biology Letters

Article Title: CIP2A promotes inflammation and exacerbates osteoarthritis by targeting CEMIP

doi: 10.1186/s11658-025-00748-0

Figure Lengend Snippet: The effect of CIP2A on cartilage is related to its downstream CEMIP. A Heatmap of differential genes identified by RNA-seq of chondrocytes transfected with si-CIP2A or si-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h, n = 3. B Volcano plot of differential genes identified by RNA-seq of chondrocytes transfected with si-CIP2A or si-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h, n = 3. C Pathway enrichment analysis of RNA-seq of chondrocytes transfected with si-CIP2A or si-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h. D NF-κB signaling pathway in GSEA enrichment analysis of RNA-seq of chondrocytes transfected with si-CIP2A or si-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h. E Pathway network of RNA-seq of chondrocytes transfected with si-CIP2A or si-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h. F Volcano plot of interacting proteins identified by CoIP-MS of chondrocytes transfected with p-CIP2A for 24 h followed by 5 ng/mL IL-1β exposure for 24 h. CIP2A was the target protein. G Network of interaction proteins identified by CoIP-MS of chondrocytes transfected with p-CIP2A for 24 h followed by 5 ng/mL IL-1β exposure for 24 h. CIP2A was the target protein. H Venn diagram depicting overlap of RNA-seq and CoIP-MS results. The Venn diagram was generated using the free online analysis tool OmicShare ( https://www.omicshare.com/tools ). I Representative WB results of endogenous CoIP validation of CIP2A and CEMIP interaction in chondrocytes, n = 3. CIP2A was the target protein of CoIP. J Representative WB results of exogenous CoIP validation of CIP2A and CEMIP interaction in chondrocytes, n = 3. Chondrocytes were transfected with the MYC-tagged CEMIP overexpression plasmid for 24 h and then treated with 5 ng/mL IL-1β for 24 h. CEMIP was the target protein of CoIP. K qRT-PCR statistical plot of CEMIP expression in chondrocytes transfected with si-CIP2A or si-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h, n = 3. L qRT-PCR statistical plot of CEMIP expression in chondrocytes transfected with p-CIP2A or p-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h, n = 3. M qRT-PCR statistical plot of CEMIP expression in chondrocytes treated with TD52 or DMSO for 24 h followed by 5 ng/mLIL-1β exposure for 24 h, n = 3. N Representative WB results of CEMIP expression in chondrocytes transfected with si-CIP2A or si-NC for 24 h followed by 5 ng/mLIL-1β exposure for 24 h, n = 3. O Representative WB results of CEMIP expression in chondrocytes transfected with p-CIP2A or p-NC for 24 h followed by 5 ng/mL IL-1β exposure for 24 h, n = 3. P Representative WB results of CEMIP expression in chondrocytes treated with TD52 or DMSO for 24 h followed by 5 ng/mL IL-1β exposure for 24 h, n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Anti-cell migration-inducing protein (CEMIP) (21129-1-AP), cartilage aggregating proteoglycan (Aggrecan) (13880-1-AP), matrix metalloproteinase (MMP) 3 (66338-1-Ig), MMP13 (18165-1-AP), β-actin (66009-1-Ig) (Supplementary Material Fig. S1) and ubiquitin (Ub) (10201-2-AP) antibodies were purchased from Proteintech (Wuhan, China).

Techniques: RNA Sequencing, Transfection, Generated, Biomarker Discovery, Over Expression, Plasmid Preparation, Quantitative RT-PCR, Expressing

Journal: Cellular & Molecular Biology Letters

Article Title: CIP2A promotes inflammation and exacerbates osteoarthritis by targeting CEMIP

doi: 10.1186/s11658-025-00748-0

Figure Lengend Snippet: Schematic diagram of the role of CIP2A in OA cartilage. In response to stimuli, CIP2A is ubiquitinated and interacts with CEMIP and PP2A. On one hand, the CIP2A/CEMIP/PP2A axis promotes ECM degradation by mediating the disorder of ECM anabolism and catabolism. On the other hand, it activates NF-κB signaling pathway, upregulates proinflammatory factors, and promotes inflammation. CIP2A leads to cartilage destruction and inflammation, accelerating OA. The schematic diagram created with BioRender.com

Article Snippet: Anti-cell migration-inducing protein (CEMIP) (21129-1-AP), cartilage aggregating proteoglycan (Aggrecan) (13880-1-AP), matrix metalloproteinase (MMP) 3 (66338-1-Ig), MMP13 (18165-1-AP), β-actin (66009-1-Ig) (Supplementary Material Fig. S1) and ubiquitin (Ub) (10201-2-AP) antibodies were purchased from Proteintech (Wuhan, China).

Techniques:

Journal: Journal for immunotherapy of cancer

Article Title: Targeting pyroptosis reverses KIAA1199-mediated immunotherapy resistance in colorectal cancer.

doi: 10.1136/jitc-2024-010000

Figure Lengend Snippet: Figure 2 KIAA1199 is related to pyroptosis. (A) Further exploration of differential gene expression between patients with CRC with high-level pyroptosis and those with low-level pyroptosis (n=633). (B, C) Comparison of KIAA1199 expression levels between patients with CRC with high-level pyroptosis and those with low-level pyroptosis through Pearson correlation analysis (B) and Gene Set Enrichment Analysis (GSEA) (C). (D) Comparison of KIAA1199 expression levels between patients with CRC with high-level pyroptosis and those with low-level pyroptosis (n=633). (E) The tumor growth curves (left) of the NC and shKIAA1199 groups were plotted. (F) The tumor growth curves (right) of the NC and shKIAA1199 groups were plotted. (G) Lactate dehydrogenase (LDH) release of serum from NC and shKIAA1199 groups. (H) Images of HCT116 and LOVO cells treated with siRNA targeting KIAA1199 are shown. The scale bar represents 50 µm, and yellow arrowheads highlight the large bubbles emerging from the plasma membrane. (I) The release of LDH from the NC and siKIAA1199 groups of HCT116 and LOVO cells was measured. (J) The release of LDH from the NC and KIAA1199 groups of HCT116 and SW480 cells was measured. All experiments were repeated at least three times. *p<0.05, **p<0.01, ***p<0.001. CRC, *p<0.05, **p<0.01, ***p<0.001. CRC, colorectal cancer.

Article Snippet: The primary antibodies, including anti- GSDMD, anti- KIAA1199, anti- DNMT1, and anti-β-actin, were purchased from Proteintech (Rosemont, Illinois, USA), whereas the anti- GSDME antibody was purchased from Abcam (Cambridge, UK).

Techniques: Gene Expression, Comparison, Expressing, Clinical Proteomics, Membrane

Journal: Journal for immunotherapy of cancer

Article Title: Targeting pyroptosis reverses KIAA1199-mediated immunotherapy resistance in colorectal cancer.

doi: 10.1136/jitc-2024-010000

Figure Lengend Snippet: Figure 3 KIAA1199 could contribute to immunotherapy resistance. (A) KIAA1199 was compared with various significant immune cell subpopulations using data from the TCGA database. (B) Comparison of responses to anti-PD1, PD-L1, or PD- L2 therapies between patients with CRC with high-level KIAA1199 and those with low-level KIAA1199 (n=633). (C) The tumor growth curves of the NC and KIAA1199 groups with PD-1 were plotted. (D) The infiltration of CD 8+T cells was analyzed by flow cytometry. (E) The infiltration of IFN-γ CD 8+T cells was analyzed by flow cytometry. (F) Comparison of CD8+ T cell infiltration between patients with CRC with KIAA1199-high and those with KIAA1199-low (n=42) through IF. (G) Comparison of OS with immunotherapy between patients with CRC with KIAA1199-high and those with KIAA1199-low (n=42). (H) Comparison of PFS with immunotherapy between patients with CRC KIAA1199-high and those with KIAA1199-low (n=42). (I) Comparison of rate of PD and DCR with immunotherapy between patients with CRC with KIAA1199-high and those with KIAA1199-low (n=42). All experiments were repeated at least three times. Fisher’s exact test was applied to panels H. *p<0.05, **p<0.01, ***p<0.001. CRC, colorectal cancer; IF, immunofluorescence; OS, overall survival; PFS, progression-free survival.

Article Snippet: The primary antibodies, including anti- GSDMD, anti- KIAA1199, anti- DNMT1, and anti-β-actin, were purchased from Proteintech (Rosemont, Illinois, USA), whereas the anti- GSDME antibody was purchased from Abcam (Cambridge, UK).

Techniques: Comparison, Flow Cytometry, Immunofluorescence

Journal: Journal for immunotherapy of cancer

Article Title: Targeting pyroptosis reverses KIAA1199-mediated immunotherapy resistance in colorectal cancer.

doi: 10.1136/jitc-2024-010000

Figure Lengend Snippet: Figure 4 KIAA1199 converts GSDME-mediated pyroptosis to apoptosis in CRC. (A) Western blot analysis for GSDMD and GSDME expression. (B) Western blot analysis was performed to assess GSDME expression in xenografts from the NC and shKIAA1199 groups. (C) Comparison of GSDME-N expression levels in HCT116 and SW480 cells was measured. (D) The percentage of apoptosis and pyroptosis cells in HCT116 cells was analyzed by flow cytometry. (E) The percentage of apoptosis and pyroptosis cells in LOVO cells was analyzed by flow cytometry. (F) The percentage of Annexin V+ PI− and Annexin V+ PI+ cells in CT26 cells was analyzed by flow cytometry. (G) Comparison of the release of LDH in HCT116, LOVO and CT26 cells was measured. (H) Comparison of GSDME-N expression levels in HCT116, LOVO and CT26 cells was measured. Western blot analysis was performed at least three times. β-actin was used as an internal control for normalization. All experiments were repeated at least three times. *p<0.05, **p<0.01, ***p<0.001. CRC, colorectal cancer; LDH, lactate dehydrogenase.

Article Snippet: The primary antibodies, including anti- GSDMD, anti- KIAA1199, anti- DNMT1, and anti-β-actin, were purchased from Proteintech (Rosemont, Illinois, USA), whereas the anti- GSDME antibody was purchased from Abcam (Cambridge, UK).

Techniques: Western Blot, Expressing, Comparison, Flow Cytometry, Control

Journal: Journal for immunotherapy of cancer

Article Title: Targeting pyroptosis reverses KIAA1199-mediated immunotherapy resistance in colorectal cancer.

doi: 10.1136/jitc-2024-010000

Figure Lengend Snippet: Figure 5 KIAA1199 inhibits pyroptosis by stabilizing DNMT1, thereby reducing the expression level of GSDME in CRC. (A, B) Co-IP experiments revealed a direct interaction between KIAA1199 and DNMT1. (C) Pull-down assays demonstrated that KIAA1199 deletion variants exhibited binding to DNMT1. (D) Comparison of DNMT1 and myc expression levels in HCT116 cells treated with cycloheximide (CHX) was measured (up). Protein bands were quantified using Image J (down). (E) Comparison of DNMT1 expression levels in HCT116 cells treated with DMSO, chloroquine, MG132 or siRNA of KIAA1199 was measured. (F) Ubiquitination assays revealed the relationship between KIAA1199 and the ubiquitination level of KIAA1199. (G) The expression levels of GSDME-N in HCT116 and LOVO cells were compared following treatment with raptinal, siRNA of KIAA1199, or overexpression of DNMT1. (H) LDH release in HCT116 and LOVO cells were compared following treatment with raptinal, siRNA of KIAA1199, or overexpression of DNMT1. Western blot analysis was performed at least three times. β-actin was used as an internal control for normalization. All experiments were repeated at least three times. *p<0.05, ***p<0.001. CO- IP, co-immunoprecipitation; CRC, colorectal cancer.

Article Snippet: The primary antibodies, including anti- GSDMD, anti- KIAA1199, anti- DNMT1, and anti-β-actin, were purchased from Proteintech (Rosemont, Illinois, USA), whereas the anti- GSDME antibody was purchased from Abcam (Cambridge, UK).

Techniques: Expressing, Co-Immunoprecipitation Assay, Binding Assay, Comparison, Ubiquitin Proteomics, Over Expression, Western Blot, Control, Immunoprecipitation

Journal: Journal for immunotherapy of cancer

Article Title: Targeting pyroptosis reverses KIAA1199-mediated immunotherapy resistance in colorectal cancer.

doi: 10.1136/jitc-2024-010000

Figure Lengend Snippet: Figure 6 Decitabine reverses the inhibition of pyroptosis mediated by KIAA1199 and sensitizes immunotherapy. (A) Comparison of DNMT1 and GSDME expression levels in HCT116 and SW480 cells treated with decitabine was measured. (B) Comparison of DNMT1 and GSDME mRNA levels in HCT116 and SW480 cells treated with DAC was measured. (C) The expression levels of GSDME in HCT116 and SW480 cells were compared following treatment with DAC, raptinal, and overexpression of KIAA1199. (D) LDH release in HCT116 and SW480 cells was compared following treatment with DAC, raptinal, and overexpression of KIAA1199. (E) The percentage of apoptosis and pyroptosis cells in HCT116 and SW480 cells following treatment with DAC, raptinal, and overexpression of KIAA1199 was analyzed by flow cytometry. (F) The tumor growth curves of the six groups were plotted. (G) Western blot analysis was performed to assess GSDME expression in xenografts from the six groups. (H) LDH release of serum from the six groups. Western blot analysis was performed at least three times. β-actin was used as an internal control for normalization. All experiments were repeated at least three times. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The primary antibodies, including anti- GSDMD, anti- KIAA1199, anti- DNMT1, and anti-β-actin, were purchased from Proteintech (Rosemont, Illinois, USA), whereas the anti- GSDME antibody was purchased from Abcam (Cambridge, UK).

Techniques: Inhibition, Comparison, Expressing, Over Expression, Flow Cytometry, Western Blot, Control

Journal: Journal for immunotherapy of cancer

Article Title: Targeting pyroptosis reverses KIAA1199-mediated immunotherapy resistance in colorectal cancer.

doi: 10.1136/jitc-2024-010000

Figure Lengend Snippet: Figure 7 Decitabine restores the release of IL-1β and the infiltration of CD8+T cells suppressed by KIAA1199. (A) IL-1β release was compared in HCT116 and LOVO cells treated with DAC, raptinal, and siRNA of KIAA1199 in tumor culture supernatants. (B) IL-1β release was compared in HCT116 and LOVO cells treated with DAC, raptinal, and overexpression of KIAA1199 in tumor culture supernatants. (C) IL-1β release of serum was analyzed by ELISA. (D) The infiltration of CD8+ T cells was analyzed by flow cytometry. (E) The tumor growth curves of the six groups were plotted. (F) LDH release of serum from the six groups. (G) Western blot analysis was performed to assess GSDME expression in xenografts from the six groups. (H) IL-1β release of serum was analyzed by ELISA. (I) The infiltration of CD8+ T cells was analyzed by flow cytometry. All experiments were repeated at least three times. *p<0.05, **p<0.01, ***p<0.001. LDH, lactate dehydrogenase.

Article Snippet: The primary antibodies, including anti- GSDMD, anti- KIAA1199, anti- DNMT1, and anti-β-actin, were purchased from Proteintech (Rosemont, Illinois, USA), whereas the anti- GSDME antibody was purchased from Abcam (Cambridge, UK).

Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Western Blot, Expressing

Journal: Journal for immunotherapy of cancer

Article Title: Targeting pyroptosis reverses KIAA1199-mediated immunotherapy resistance in colorectal cancer.

doi: 10.1136/jitc-2024-010000

Figure Lengend Snippet: Figure 4 KIAA1199 converts GSDME-mediated pyroptosis to apoptosis in CRC. (A) Western blot analysis for GSDMD and GSDME expression. (B) Western blot analysis was performed to assess GSDME expression in xenografts from the NC and shKIAA1199 groups. (C) Comparison of GSDME-N expression levels in HCT116 and SW480 cells was measured. (D) The percentage of apoptosis and pyroptosis cells in HCT116 cells was analyzed by flow cytometry. (E) The percentage of apoptosis and pyroptosis cells in LOVO cells was analyzed by flow cytometry. (F) The percentage of Annexin V+ PI− and Annexin V+ PI+ cells in CT26 cells was analyzed by flow cytometry. (G) Comparison of the release of LDH in HCT116, LOVO and CT26 cells was measured. (H) Comparison of GSDME-N expression levels in HCT116, LOVO and CT26 cells was measured. Western blot analysis was performed at least three times. β-actin was used as an internal control for normalization. All experiments were repeated at least three times. *p<0.05, **p<0.01, ***p<0.001. CRC, colorectal cancer; LDH, lactate dehydrogenase.

Article Snippet: The primary antibodies, including anti- GSDMD, anti- KIAA1199, anti- DNMT1, and anti-β-actin, were purchased from Proteintech (Rosemont, Illinois, USA), whereas the anti- GSDME antibody was purchased from Abcam (Cambridge, UK).

Techniques: Western Blot, Expressing, Comparison, Flow Cytometry, Control