Fig 1: Low SPINK4 levels were associated with significantly decreased overall survival in CRC patients from a GSE24551, b GSE39582, c TCGA and d our study cohort. Low SPINK4 levels were associated with significantly decreased disease-free survival rates in CRC patients from e our study cohort
Fig 2: A RBP4+NTS+ cancer cell subset is unique to left-sided CRC.(A) The t-SNE plot that showed the distribution of cancer cell lineages (yellow, n = 2196 cells) within the atlas. Cancer cell populations were reclustered into 9 subclusters (color coding). (B) Top 5 marker genes of 15 major cell types identified in this profile. (C) Annotation by left-sided and right-sided CRC cells. (D) The fraction of cells that originated from left-sided and right-sided CRC samples for 9 subgroups identified in this profile. (E) Kaplan-Meier survival curves of OS based on HSPA1A, GOLGB1, AGR3, MUC5AC, NTS, SPINK4, CLCA1, OLFM4, and PIGR expression using the online bioinformatics tool Kaplan-Meier Plotter. (F) Violin plots and immunochemistry display the distribution of expression of AGR2, AGR3, MUC5AC, SPINK4, TFF1, and TFF2 between left-sided and right-sided CRC. Scale bars: 100 µm.
Fig 3: SPINK4 expression was higher in pancreatic adenocarcinoma (PAAD) and gastric adenocarcinoma (STAD) tissues than in the corresponding normal tissues, as evidenced by analysis of the GEPIA database. BLCA, Bladder Urothelial Carcinoma; BRCA, Breast invasive carcinoma; CESC, Cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, Cholangio carcinoma; COAD, Colon adenocarcinoma; ESCA, Esophageal carcinoma; HNSC, Head and Neck squamous cell carcinoma; KICH, Kidney Chromophobe; KIRC, Kidney renal clear cell carcinoma; KIRP, Kidney renal papillary cell carcinoma; LIHC, Liver hepatocellular carcinoma; LUAD, Lung adenocarcinoma; LUSC, Lung squamous cell carcinoma; PAAD, Pancreatic adenocarcinoma; PCPG, Pheochromocytoma and Paraganglioma; PRAD, Prostate adenocarcinoma; READ, Rectum adenocarcinoma; SARC, Sarcoma; SKCM, Skin Cutaneous Melanoma; STAD, gastric adenocarcinoma; THCA, Thyroid carcinoma; THYM, Thymoma; UCEC, Uterine Corpus Endometrial Carcinoma
Fig 4: Representative images of SPINK4 immunohistochemical staining in CRC tissues from our study cohort. a Negative SPINK4 expression in CRC tissue (magnification × 200); b low SPINK4 expression in CRC tissue (magnification × 200); c moderate-strong SPINK4 expression in CRC tissue (magnification × 200); d strong SPINK4 expression in normal tissue. (magnification × 200); e Representative images of SPINK4 protein expression in sections of nonneoplastic mucosa adjacent to tumors (magnification × 200); f Representative images of SPINK4 protein expression in sections of nonneoplastic mucosa adjacent to tumors (magnification × 400)
Fig 5: SPINK4 is downregulated in CRC. SPINK4 expression was significantly decreased in CRC tissues compared with that in paired adjacent normal tissues at the mRNA level in the (a) GSE32323 dataset and the (b) GSE39582 dataset and at the protein level in (c) our own cohort. d SPINK4 expression was decreased in adenoma tissues compared to that in adjacent normal mucosal tissues in GSE3880. e SPINK4 expression was similar between adenoma and carcinoma tissues in GSE3880. Validation of the expression of SPINK4 at the protein level in (f) normal and (g) CRC tissues by the Human Protein Atlas database (IHC staining)
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