Fig. 1: KCTD9 is downregulated in CRC.
From: KCTD9 inhibits the Wnt/β-catenin pathway by decreasing the level of β-catenin in colorectal cancer
A Venn diagram showing intersecting downregulated differentially expressed genes (DEGs), as assessed by RNA sequencing (RNA-seq) results of three tissue samples from our hospital, and other public datasets: The Cancer Genome Atlas (TCGA)-COAD, GSE39582 and GSE75970. RNAseq_DEGs_Down: DEGs with consistently descending expressed trends among the adjacent normal colonic mucosa, primary colon adenocarcinoma and distant liver metastasis were identified from the RNA-seq data of three tissue samples in our hospital. COAD_DEGs_Down: DEGs downregulated between COAD vs. Normal groups based on TCGA-COAD data. COAD_DEGs_OS_Sig: DEGs significantly associated with overall survival (OS) of TCGA-COAD data predicted by both univariable cox and log-rank methods. GEO_DEGs_Down: Intersected DEGs downregulated between COAD/CRC vs. Normal groups based on GSE39582 and GSE75970 data. B Comparative expression of KCTD9 in normal colonic epithelium, primary and metastatic CRC tissues determined using the TNMplot web tool. C Western blot analysis of KCTD9 expression in 12 matched pairs of human CRC tissues and adjacent normal colonic tissues collected during resection (n = 12). β-actin was used as a loading control throughout. Relative density were quantified by using ImageJ software. D, E Tissue microarrays containing 80 pairs of human CRC tissues and adjacent normal colon tissues were analyzed for KCTD9 expression using immunohistochemical (IHC) staining. Representative staining in normal colon gland epithelium and adenocarcinoma tissues (D) (n = 80). Quantification of IHC staining in D as immunoreactive scores (IRS) E mean ± SD, n = 80, two-tailed Student’s t test, ***p < 0.001. Comparative analysis of KCTD9 expression in the normal human colon mucosal epithelial cell line NCM460 and a panel of CRC-derived cell lines using qRT-PCR (F) and Western blotting (G). G Is represent three independent experiments (top) and relative density were quantified by using ImageJ software (bottom). F, G Are mean ± SD; n = 3 independent experiments, one-way ANOVA with Tukey’s multiple comparison post-test, ns not significant, **p < 0.01, ***p < 0.001.
