colorectal cancer tissue microarray  (ZHUOLI IMAGING TECHNOLOGY CO LTD)

Journal: Cancer Genomics & Proteomics

Article Title: Targeting Bmi1 for Enhancing Anoikis Sensitivity and Inhibiting Metastasis in Colorectal Cancer

doi: 10.21873/cgp.20469

Figure Lengend Snippet: The level of Bmi1 expression dictates the metastatic potential of colorectal cancer (CRC) cells. (A) Formalin-fixed paraffin-embedded samples from four pairs of CRC patients were stained with the anti-Bmi1 antibody (magnification, 200×). (B) Bmi1 expression was evaluated via western blotting using anti-Bmi1 antibodies in SW480 and SW620 cells. (C) Migration of SW620 cells and Bmi1-depleted cells in zebrafish. (D) Expression of Bmi1 was analyzed via western blotting using anti-Bmi1 antibodies in suspended SW620 cells at specified time points.

Article Snippet: A human colorectal cancer tissue microarray (CDA3, Biomax Inc. and Super Bio Chips Laboratories, Seoul, Republic of Korea) was utilized.

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Staining, Western Blot, Migration

Journal: Cancer Genomics & Proteomics

Article Title: Targeting Bmi1 for Enhancing Anoikis Sensitivity and Inhibiting Metastasis in Colorectal Cancer

doi: 10.21873/cgp.20469

Figure Lengend Snippet: Bmi1 regulates anoikis resistance in colorectal cancer (CRC) cells. (A) Migration of SW620 cells and Bmi1-depleted cells was assessed using a Transwell assay after 24 h of growth in suspension. (B) The percentage of apoptotic SW620 cells and Bmi1-depleted cells at specified time points in suspension was determined by dual staining with annexin V-FITC and PI. The experiment was replicated thrice, and between-group comparisons were performed using a one-way ANOVA (*p<0.05). (C) The percentage of apoptotic SW620 cells treated with the Bmi1 inhibitor PTC209 at specified time points in suspension was determined by dual staining with annexin V-FITC and PI. The experiments were replicated thrice, and between-group comparisons were performed using a one-way ANOVA (*p<0.05). (D) Expression levels of Bmi1 and cleaved caspase-3 were assessed via western blotting after 48 h of suspension.

Article Snippet: A human colorectal cancer tissue microarray (CDA3, Biomax Inc. and Super Bio Chips Laboratories, Seoul, Republic of Korea) was utilized.

Techniques: Migration, Transwell Assay, Suspension, Staining, Expressing, Western Blot

Journal: Cancer Genomics & Proteomics

Article Title: Targeting Bmi1 for Enhancing Anoikis Sensitivity and Inhibiting Metastasis in Colorectal Cancer

doi: 10.21873/cgp.20469

Figure Lengend Snippet: Correlation between Bmi1, MDK and clinicopathological parameters. in colorectal cancer.

Article Snippet: A human colorectal cancer tissue microarray (CDA3, Biomax Inc. and Super Bio Chips Laboratories, Seoul, Republic of Korea) was utilized.

Techniques:

Journal: Advanced Science

Article Title: Protein Kinase STK24 Promotes Tumor Immune Evasion via the AKT‐PD‐L1 Axis

doi: 10.1002/advs.202304342

Figure Lengend Snippet: STK24 expression is upregulated in multiple cancer types and correlated with poor prognosis. A–C) Statistical analysis of STK24 protein expression levels determined by IHC staining of the adjacent‐tumor tissues and tumor tissues from the patients with colorectal cancer (CRC) (A), lung adenocarcinoma (LUAD) (B), or pancreatic adenocarcinoma (PAAD) (C). D–F) Statistical analysis of STK24 protein expression levels determined by IHC staining of the adjacent‐tumor tissues and tumor tissues from the same patient with CRC ((D) n = 60), LUAD ((E) n = 81), or PAAD ((F) n = 90). G–I) Kaplan–Meier curves of overall survival in the set of patients with G) CRC, H) LUAD, I) or PAAD based on STK24 protein expression level detected in the tumor tissues. J–L) Comparison of STK24 gene expression levels in tumor tissues or normal tissues from patients with LUAD (J), lung squamous cell carcinoma (LUSC) (K), and cholangio carcinoma (CHOL) (L) based on the HPA database. M,N) Comparison of STK24 expression levels in tumor tissues or normal tissues from patients with pancreatic adenocarcinoma (PAAD) (M) and rectum adenocarcinoma (READ) (N) based on the GEPIA database. Results are presented as mean ± SEM. * p <0.05, ** p <0.01, **** p <0.0001. P values were calculated by unpaired Student's t ‐tests in (A–F) and (J–L) and log‐rank test in (G–I). See also Figure (Supporting Information).

Article Snippet: Immunohistochemistry staining of the tissue microarray (TMA) of colorectal cancer was performed by Servicebio.

Techniques: Expressing, Immunohistochemistry, Comparison, Gene Expression

Journal: Human pathology

Article Title: Validation of genetic classifiers derived from mouse and human tumors to identify molecular subtypes of colorectal cancer

doi: 10.1016/j.humpath.2021.10.002

Figure Lengend Snippet: (A) CRC tissue microarray (CRC TMA) was stained for four previously established IHC classifiers (MUC2, TFF3, CFTR, and ZEB1). Qualitative scoring of epithelial cells was performed using a tiered system for MUC2, TFF3 and CFTR (0, no staining; 1, low intensity; 2, moderate intensity; and 3, high intensity). ZEB1 was scored based on the presence or absence of epithelial cell staining. Representative images of each score are shown on the right, unless the score was not observed (NA, not applicable). On the left, graphs show the percent of cancers for each score tier for MUC2, TFF3, CFTR, and for the presence/absence of ZEB1. For whole core images, scale bar represents 300 μm. For magnified images, scale bar represents 100 μm. (B) The classification of the cancers based on IHC classifiers in (A).

Article Snippet: Colorectal cancer tissue microarray (CRC TMA) A formalin-fixed paraffin-embedded (FFPE) CRC TMA was developed by the University of Wisconsin Carbone Cancer Center Translational Science Biocore (TSB) under an IRB-approved protocol (2016–0934) as previously described [ 15 ].

Techniques: Microarray, Staining

Journal: Human pathology

Article Title: Validation of genetic classifiers derived from mouse and human tumors to identify molecular subtypes of colorectal cancer

doi: 10.1016/j.humpath.2021.10.002

Figure Lengend Snippet: (A) Colorectal cancer tissue microarray (CRC TMA) was stained for previously established IHC classifiers (CDX2, FRMD6, HTR2B, cytokeratin, and ZEB1) to distinguish CMS2/3 from CMS4. CMS1 was determined solely by mismatch repair deficiency (dMMR). Qualitative scoring of epithelial cells was performed using a tiered system for CDX2, FRMD6, HTR2B, cytokeratin (0, no staining; 1, low intensity; 2, moderate intensity; and 3, high intensitywith a score of 0 or 1 being classified as low and a score of 2 or 3 being scored as high). ZEB1 is shown in Figure 1A. Representative images of each scoring tier are shown on the right for CDX2, FRMD6, HTR2B and cytokeratin, unless the score was not observed (NA, not applicable). On the left, the percent of cancers for each score tier is shown. CMS2/3 is expected to have low CDX2, high FRMD6, high HTR2B and the absence of ZEB1 in epithelial cells. CMS4 is expected to have high CDX2, low FRMD6, low HTR2B and the presences of ZEB1 in epithelial cells. For whole core images, scale bar represents 300 μm. For magnified images, scale bar represents 100 μm. (B) The distribution of cancers on our CRC TMA amongst the CMS classes based on IHC classifiers in Figure 2A. (C) There was no difference in disease-free survival (DFS) or overall survival (OS) based on the CMS classification of our CRC TMA patient cohort. (D) When assessed by cancer stage, there was no difference in DFS or OS for CMS1 cancers (left). CMS2/3 cancers (right) showed the expected trend of a decrease in survival with an increase in cancer stage for both OS and DFS.

Article Snippet: Colorectal cancer tissue microarray (CRC TMA) A formalin-fixed paraffin-embedded (FFPE) CRC TMA was developed by the University of Wisconsin Carbone Cancer Center Translational Science Biocore (TSB) under an IRB-approved protocol (2016–0934) as previously described [ 15 ].

Techniques: Microarray, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Neuromedin U secreted by colorectal cancer cells promotes a tumour-supporting microenvironment

doi: 10.1186/s12964-022-01003-1

Figure Lengend Snippet: Human CRC tissues—IHC and TCGA data analysis. a Tissue samples (grade 1) collected on tissue microarrays stained with anti-NMU antibodies (IHC) and analysed under the microscope (zoom 20x, field 0.3 mm 2 ). b Percent of all analysed cores with different levels of NMU staining (Fisher’s exact test). c Percent of NMU-positive epithelial cells in the analysed CRC tissues, grade 1 (Wilcoxon matched-pairs signed rank test). NAT—normal adjacent tissue. d Changes in NMU and NMUR s expression in CRC tissues compared to normal samples based on the expression levels extracted from TCGA RNA sequencing dataset (published here ). e Ordered patient data with survival data were divided into NMUR low (n = 260) and NMUR high (n = 260) expression groups according to the median values, and survival rates were compared between the groups by performing a Kaplan–Meier analysis (log-rank (Mantel–Cox) test, χ 2 = 6.559, df = 1, p = 0.0104). NMUR1 expression was extracted from TCGA RNA sequencing dataset and compared among CRC tumours with different f MSI statuses (Kruskal–Wallis test, p < 0.0001, Dunn’s multiple comparisons test, *** p = 0.0008; **** p < 0.0001), g M stages (Mann–Whitney test, two-tailed, p = 0.4419), h N stages (Mann–Whitney test, two-tailed, p = 0.0079), and i VEFGC expression between groups with NMUR1 low and NMUR1 high levels according to the median values (Mann–Whitney test, two-tailed, **** p < 0.0001). The boxes represent the interquartile ranges, the horizontal lines in the boxes represent the medians, and the whiskers represent the minimum and maximum values

Article Snippet: A rabbit anti-NMU antibody (Sigma–Aldrich, Saint Louis, MO, USA) was used, and immunohistochemical staining was performed on commercially available colorectal cancer tissue microarrays (TissueArray.Com LLC, Derwood, MD, US previously US Biomax).

Techniques: Staining, Microscopy, Expressing, RNA Sequencing Assay, MANN-WHITNEY, Two Tailed Test

Journal: bioRxiv

Article Title: TMIGD1, a putative tumor suppressor, induces G2-M cell cycle checkpoint arrest in colon cancer cells

doi: 10.1101/2020.06.06.138057

Figure Lengend Snippet: ( A ) Two human colorectal cancer tissue microarray slides consist of 172 tissue samples were stained with anti-TMIGD1 antibody. Staining was scored as 0 (negative, <5% cells positive), 1+ (6-25% cells positive), 2+ (26-50% cells positive), and 3+ (>50% cells positive). The tumor differentiation was graded morphologically (grade 1, 2 and 3 as well, moderately and poorly differentiated). ( B ) The average staining intensities were compared using ANOVA with Tukey post-hoc test per tumor grade. ( C ) The average staining intensities were compared using ANOVA with Tukey post-hoc test per tumor differentiation. ( D ) Western blot analysis of TMIGD1 expression in human renal and colorectal cancer cell lines. ( E ) qPCR analysis of TMIGD1 expression in CRC cell lines (HT29, HCT116 and RKO). Human kidney epithelial cells, HK2 was used as a positive control.

Article Snippet: Two human colorectal cancer tissue microarray slides (US Biomax, catalog numbers BC05012a and BC05118a) consist of 172 tissue samples (72 on BC05012a and 100 on BC05118a) were stained with anti-TMIGD1 antibody.

Techniques: Microarray, Staining, Western Blot, Expressing, Positive Control

Journal: Cancer Cell International

Article Title: Capicua suppresses colorectal cancer progression via repression of ETV4 expression

doi: 10.1186/s12935-020-1111-8

Figure Lengend Snippet: ETV4 promotes CRC progression. a Analysis of the TCGA dataset for levels of ETV1, ETV4 , and ETV5 in normal colon (NC) and CRC samples of four different clinicopathological stages (I, II, III, and IV). b The graph for the proportion of normal colon ( n = 20) and CRC ( n = 188) tissue samples with different ETV4 expression scores (−, +, ++). The CRC tissue microarray was subjected to immunohistochemical analysis of ETV4 protein levels. c Western blot analyses of ETV4 levels in ETV4-overexpressing (ETV4 OE) and ETV4-knockdown (shETV4) CRC cells (HCT116 and HT29). d Cell proliferation assay of ETV4-overexpressing (ETV4 OE) and ETV4-knockdown (shETV4) CRC cells. e In vivo subcutaneous tumor growth curves of control (scramble) and ETV4-knockdown (shETV4) HCT116 cells. n = 6 per each group. The inset image is a representative image of xenograft tumors dissected from the mice after the last measurement of tumor size. The right panel represents a graph for average weights of the dissected tumors. f , g Matrigel invasion ( f ) and trans-well migration ( g ) assay of control, ETV4-overexpressing, and ETV4-knockdown CRC cells. The bottom panels represent the bar graph for quantification of cell invasiveness (f) and cell migration (g) , respectively. Three independent experiments were performed. All error bars indicate s.e.m. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: The colorectal cancer tissue microarray (CO2085b) was purchased from Biomax (MD, USA).

Techniques: Expressing, Microarray, Immunohistochemical staining, Western Blot, Knockdown, Proliferation Assay, In Vivo, Control, Migration

Journal: Cancer Cell International

Article Title: Capicua suppresses colorectal cancer progression via repression of ETV4 expression

doi: 10.1186/s12935-020-1111-8

Figure Lengend Snippet: Loss of CIC in colorectal cancer. a Analysis of the TCGA dataset for CIC mRNA levels in normal colon (NC) and primary colorectal tumor samples. The numbers in parentheses indicate the number of subjects in each group. * P < 0.05. b Analysis of the TCGA dataset for CIC mRNA levels in normal colon (NC) and CRC samples of four different clinicopathological stages (I, II, III, and IV). The numbers in parentheses indicate the number of subjects in each group. * P < 0.05. c Analysis of CIC protein levels in normal colon and CRC tissues by immunohistochemistry. d Western blot analysis for CIC and ETV4 levels in normal colon (N) and CRC (T) samples from the same patient with CRC. Samples from four CRC patients were subjected to this experiment

Article Snippet: The colorectal cancer tissue microarray (CO2085b) was purchased from Biomax (MD, USA).

Techniques: Immunohistochemistry, Western Blot

Journal: EBioMedicine

Article Title: Type Iγ phosphatidylinositol phosphate kinase promotes tumor growth by facilitating Warburg effect in colorectal cancer

doi: 10.1016/j.ebiom.2019.05.015

Figure Lengend Snippet: High PIPKIγ expression level predicts a poor clinical outcome in colorectal cancer. (A) Real-time qPCR analysis of the mRNA level of PIPKIγ in colorectal cancer cell lines and the normal colonic epithelial cell NCM460. (B) Cell lysates of indicated cells were collected for immunoblotting analyses using PIPKIγ antibody; β-actin was loaded as a control. (C) IHC analysis was performed in a tissue microarray containing 76 matched tumor and non-tumor colorectal cancer tissues. Representative images of PIPKIγ and its expression intensity in non-tumor and tumor tissues were shown. Scale bar: 100 μm. (D) Kaplan-Meier analyses of overall survival of individuals with colorectal cancer based on PIPKIγ protein expression level. (E) Kaplan-Meier analyses of overall survival in colon adenocarcinoma (COAD) and rectal adenocarcinoma (READ) patients in the Cancer Genome Atlas (TCGA) cohort. The patients were dichotomously categorized on the basis of median PIPKIγ value into 2 groups. Subgroups were compared with the use of the log-rank test. * P < .05; ** P < .01; *** P < .001.

Article Snippet: The colorectal cancer tissue microarray used in this study was purchased from Zhuoli Biotech (#COC1504, http://www.zhuolibiotech.com/ , Shanghai, China).

Techniques: Expressing, Western Blot, Control, Microarray

Journal: EBioMedicine

Article Title: Type Iγ phosphatidylinositol phosphate kinase promotes tumor growth by facilitating Warburg effect in colorectal cancer

doi: 10.1016/j.ebiom.2019.05.015

Figure Lengend Snippet: PIPKIγ promotes colorectal cancer cell proliferation in vitro and tumor growth in vivo. (A) COAD samples derived from TCGA cohort was categorized into 2 groups (high versus low) based on median PIPKIγ value. Gene set enrichment analysis (GSEA) was performed to discover the difference between 2 groups. False discovery rate (FDR) was set at 0.25. NES represents normalized enrichment score. (B) Validation of pan-PIPKIγ knockdown and ectopic expression of mutant-PIPKIγ_i2 (resistant to PIPKIγ shRNA) in SW480 and LOVO cells using Western blotting. (C, D) The influence of PIPKIγ on colorectal cancer in vitro cell proliferation was determined by CCK-8 (C) and colony formation (D) assays, respectively. (E) sh-Ctrl, sh-PIPKIγ-1, and sh-PIPKIγ-1 + mPIPKIγ1_i2 SW480 cells were injected subcutaneously into the left forelimb of nude mice ( n = 6 per group). Four weeks later, mice were sacrificed and tumor weights in each group were shown. (F) IHC analysis of PIPKIγ and PCNA expression from indicated subcutaneous xenograft. * P < .05 and ** P < .01.

Article Snippet: The colorectal cancer tissue microarray used in this study was purchased from Zhuoli Biotech (#COC1504, http://www.zhuolibiotech.com/ , Shanghai, China).

Techniques: In Vitro, In Vivo, Derivative Assay, Biomarker Discovery, Knockdown, Expressing, Mutagenesis, shRNA, Western Blot, CCK-8 Assay, Injection