图1 人结直肠癌患者中DSPP的表达与奥沙利铂敏感性的相关性

Fig.1 DSPP correlates with oxaliplatin responsiveness in colorectal cancer (CRC). A: Immunohistochemical (IHC) images showing DSPP expression in tumors from oxaliplatin (L-OHP)‑sensitive and ‑resistant CRC patients (***P<0.001). B: Quantification of DSPP IHC scores in L-OHP-sensitive and -resistant CRC tumors (n=30). C: CCK-8 assay for assessing viability of L-OHP-resistant HCT8 cell line (n=4). D: Western blotting of DSPP in L-OHP-sensitive HCT8 and L-OHP-resistant HCT8 cells. E: Western blotting confirming stable DSPP knockout in HCT8 cells. F: CCK-8 assay showing increased L-OHP sensitivity in DSPP-knockout HCT8 cells (n=4).

图2 DSPP促进结直肠癌类器官和细胞系对奥沙利铂耐药

Fig.2 DSPP depletion enhances oxaliplatin resistance in human CRC organoids and cell lines. A: Representative schematic of the establishment and characterization of patient-derived tumor organoids (PDTOs). Scale bar=100 μm. B: HE and immunofluorescence staining showing DSPP expression in edited PDTOs. Scale bar=100 μm. C: CCK-8 assay showing significantly increased L-OHP sensitivity in DSPP-knockout PDTOs (n=4). D: Western blotting showing the efficiency of DSPP knockout in HCT116 and SW620 cells. E: CCK-8 assays showing increased L-OHP sensitivity in DSPP knockout HCT116 and SW620 cells (n=4).

图3 靶向DSPP增强奥沙利铂在裸鼠模型和人源肿瘤异种移植模型中的疗效

Fig.3 Targeting DSPP augments oxaliplatin efficacy in xenograft and PDX models.A: Representative subcutaneous xenograft tumors and IHC demonstrating the efficacy of combining L-OHP with a DSPP-targeting therapy (n=5). Scale bar=50 μm. B: Endpoint tumor weights in each group (n=5). C: Tumor growth curves over time in each group (n=5). D: Schematic diagram of patient-derived xenograft (PDX) establishment and treatment regimens. E, G, I: Representative macroscopic tumors, HE staining, and DSPP IHC from 3 independent PDX cases (n=5 or 6). Scale bar=200 μm. F, H, J: Tumor growth curves and tumor weights in the corresponding PDX cases. *P<0.05, **P<0.01, ***P<0.001.

图4 DSPP与整合素αvβ3相互作用激活MAPK信号通路

Fig.4 DSPP interacts with integrin αvβ3 and modulates MAPK signaling. A, B: Co-immunoprecipitation (Co-IP) and immunofluorescence images showing the interaction and subcellular co-localization of DSPP and integrin αvβ3 in CRC cells. Scale bar=10 μm. C: Immunofluorescence images showing co-localization of DSPP and αvβ3 in CRC tissues. Scale bar=50 μm. D: IHC showing the correlation between DSPP and integrin αvβ3 expression in human CRC specimens. Scale bar=50 μm. E, F: Western blotting showing that DSPP regulates MAPK/ERK and p53 pathways in the indicated CRC cell lines, and the integrin-targeted inhibitor Cyclo (at a concentration of 38.33 ng/mL) reverses DSPP-induced upregulation of ERK and P53 phosphorylation levels.

图5 PDX模型证实靶向DSPP、整合素αvβ3增强结直肠癌PDX奥沙利铂化疗敏感性

Fig.5 Targeting DSPP or integrin αvβ3 enhances oxaliplatin efficacy in CRC PDX models. A: Schematic diagram of the PDX experimental design for evaluating L-OHP combined with DSPP-targeting antibody or αvβ3 inhibitor Cyclo(-RGDfK). B, E: Representative macroscopic tumors, HE staining, and DSPP IHC from two independent PDX cases (n=4-6). Scale bar=100 μm. C, F: Tumor growth curves for the indicated treatment groups (n=4-6). D, G: Endpoint tumor weights in each group (n=4-6). H: Body-weight monitoring of PDX mice across different treatment groups, showing no significant toxicity (n=4-6). I: HE staining of major organs (intestine, liver, heart, and spleen) from PDX mice across the treatment groups. Scale bar=50 μm. *P<0.05, **P<0.01, ***P<0.001.