Expression In Colorectal Carcinoma Research Articles

Evaluation of PD-L1 Expression in Colorectal Carcinomas by Comparing Scoring Methods and Their Significance in Relation to Clinicopathologic Parameters.

This study aims to evaluate PD-L1 expression in colorectal carcinomas (CRCs) by using the tumor proportion score (TPS) and the combined positive score (CPS), and to investigate whether there is a correlation with clinicopathologic features. A cross-sectional study was conducted that included samples from patients with colorectal adenocarcinoma treated with colon resection and rectal resection after neoadjuvant radio- and chemotherapy at the Department of Abdominal Surgery at Požega Hospital in the period from 2017 to 2022. The study included 102 tumor tissue samples from patients after resection and the pathohistological diagnosis of adenocarcinoma. In our study, the PD-L1 positivity rate after the TPS was 42 (41%) samples, and after the CPS, 97 (95%) of them (p < 0.001). The positive expression of PD-L1 in tumor cells using the TPS method showed a statistically significant association with adenocarcinoma (TPS ≥ 10-50% and ≥50%). There were significantly more that were moderately differentiated, with TPS ≥ 50%, and those poorly differentiated had values ≥ 10-50%. There were significantly more patients with a status of more than one positive lymph node with TPS values ≥ 10-50%. Patients without metastases in the lymph nodes are significantly more likely to have CPS values > 50%, compared with other lymph node statuses. These results suggest that the total number of PD-L1-expressing cells, including tumor and immune cells, is a more sensitive biomarker than the number of PD-L1-expressing tumor cells alone in CRC.

PD-L1 Expression in Colorectal Carcinoma Correlates with the Immune Microenvironment.

Colorectal carcinoma (CRC) is a common malignancy, with its diverse clinical, pathological, and molecular features. The immune microenvironment of a tumor comprises of interplay between various cells and molecules, and has a significant role in deciding the tumor behavior and overall prognosis. PD-L1 (programmed cell death ligand-1) has been implicated in the regulation of the tumor immune microenvironment (TIME). There is limited data regarding the correlation of PD-L1 expression with immune cell profile in CRCs, especially in the Indian setting. The study aimed to assess the PD-L1 expression in CRC tumor cells and its association with TIME, mismatch repair (MMR), and various other clinicopathological parameters. This is a hospital-based, cross-sectional observational study. PD-L1 expression was assessed at the protein level by immunohistochemistry and mRNA level by qRT-PCR. Immune cell markers (CD4, CD8, CD20, FOXP3, and CD163) were interpreted using the ImageJ Fiji platform. Of the 104 cases, 21% were PD-L1 positive and were more common in right-sided CRCs. PD-L1 positive cases showed significantly higher concentrations of all T-cell subsets (CD4+ , CD8+ , and FOXP3+), CD20+ B-cells, and CD163+ macrophages were noted. No statistical significance was seen between PD-L1 expression with clinical profile, pathological subtype, grade or stage, mismatch repair status (proficient vs deficient), and survival. The present study showed a relatively lower frequency of PD-L1 in CRC from the Eastern Indian cohort. The immune cell concentration in the present study was calculated using image analysis-based objectivised methods. Significant correlation of PD-L1 expression in tumor cells with the tumor-infiltrating immune cells indicated its crucial role in the pathobiology of CRC especially by regulating the TIME. Considering the therapeutic implication of PD-L1 in various malignancies, it may be one of the crucial therapeutic targets in a proportion of cases.

Beta Catenin Expression in Colorectal Carcinoma and its relation to survival and prognostic factors

Beta Catenin Expression in Colorectal Carcinoma and its relation to survival and prognostic factors

Role of HER2/neu in Colorectal Carcinoma: A Cross-sectional Study from a Tertiary Care Centre in West Bengal, India

Introduction: In many tumours, Human Epidermal Growth Factor Receptor 2 (HER2) expression and HER2 amplification are studied as therapeutic and prognostic factors. The incidence of HER2/neu expression and its consequences in patients with colorectal carcinoma has shown conflicting evidence. Aim: To evaluate the utility of HER2/neu expression in colorectal carcinoma and to find any association between HER2/neu expression, histological grade, and stage of the tumour. Materials and Methods: This cross-sectional study was conducted on 50 cases of colorectal carcinoma. Patients undergoing surgery for colorectal tumours were selected over an 18-month period (January 2020 to June 2021) in a tertiary care hospital in Kolkata, West Bengal, India. Various clinical and demographic parameters were obtained from the patients, and the specimens were subjected to Immunohistochemical (IHC) staining for HER2/neu. The results were analysed using Statistical Package for the Social Sciences (SPSS)software version 25.0 (IBM, Armonk, New York, USA). Results: On HER2/neu IHC examination, 30 cases (60%) showed a 3+ score, 4 cases (8%) showed a 2+ score, 7 cases (14%) showed a 1+ score, and the remaining 9 cases (18%) showed a score of 0. HER2/neu expression was found in 30 cases (60%) of colorectal carcinoma. HER2/neu expression showed a significant association with the histological grade of the tumour (p-value=0.020). HER2/neu expression also showed a significant association (p-value&lt;0.001) with histological subtypes of the tumour. Conclusion: This study revealed a significant association between HER2/neu expression and histological subtypes and grade of the tumour. Moderately differentiated adenocarcinoma showed the highest expression, while mucinous adenocarcinoma and undifferentiated carcinoma showed minimal or absent HER2/ neu expression. Thus, HER2/neu can serve as a prognostic and therapeutic marker in colorectal carcinoma. In conclusion, surgery alone is not curative for patients with colorectal cancer, and additional forms of management may be required to improve patient survival. In such cases, HER2/neu expression can guide new adjuvant therapy.

Evaluation of HHLA2 and CD8 Immunohistochemical Expression in Colorectal Carcinoma and Their Prognostic Significance.

Colorectal carcinoma (CRC) is the third most common malignancy worldwide. Human endogenous retrovirus H long terminal repeat-associating protein 2 (HHLA2) is a novel immune checkpoint molecule. The association between HHLA2 expression and clinicopathological features and its prognostic significance in CRC patients are still controversial. The aim of this study is to evaluate the prognostic value of immunohistochemical (IHC) expression of HHLA2 and CD8 in CRC. This retrospective study included 134 cases diagnosed with primary CRC at the Gastrointestinal Surgery Center (GISC) department, Mansoura Faculty of Medicine, during the period from December 2014 to December 2018. Clinicopathological and survival data were collected. IHC for HHLA2 and CD8 was performed, and they were correlated with clinicopathological parameters and patient prognosis. Among 134 CRC cases, high HHLA2 expression was detected in 73 (54.5%). High HHLA2 expression was significantly related to the depth of invasion (P = 0.005*), lymph node metastasis (P = 0.01*), tumor stage )P = 0.002*), and distant recurrence )P = 0.012*). Multivariate analysis spotted HHLA2 high expression as an independent prognostic predictor for OS in CRC (P = 0.03*) and DFS (P = 0.008*). CD8 shows a significant correlation with tumor infiltrating lymphocytes (TILs) (P ≤ 0.001*), absence of metastasis ((P = 0.029*), absence of tumor deposits (P=0.014*). However, CD8 shows no significant association with survival or HHLA2. HHLA2 is an independent prognostic factor for the overall survival and disease free survival of CRC patients and can predict poor prognosis in CRC patients.

Immunohistochemical analysis of cytokeratin 7 and 20 expression in colorectal carcinoma and its correlation with histopathological grading of tumour

Background: Colorectal carcinoma is a malignant epithelial tumor originating in the large bowel. More than 90% of colorectal carcinomas (CRC) are adenocarcinomas. This being graded morphologically at H&amp;E stain in histopathology as well-differentiated, moderately differentiated, and poorly differentiated. Aims and Objectives: The aim of this study is to assess any association with different histopathological types and extent of differentiation of CRC with change in expression of CK 7 and Cytokeratin 20 (CK20). Materials and Methods: In the present study, total 59 patients has been selected by the predetermined sampling techniques. The specimen was collected and the necessary processing was done for H&amp;E stain and for immunohistochemistry for CK 20 and CK 7 profile. Results: This study showed that most of the well-differentiated colorectal carcinoma are CK20 (+ve)/Cytokeratin 7 (CK7) (-ve) and poorly differentiated CRC are CK20 (+) and/CK7 (+) and also CK20 (−ve)/CK7 (+ve). Conclusion: Hence, from this study, it is evident that the expression of CK 20 positivity decreasing and CK7 positivity increasing in respect of progression of well differentiated to poorly differentiated colorectal carcinoma, that is, there is a relationship of expression of CK20 and CK7 profile with the histological grading of colorectal carcinoma. Furthermore, the expression profile of CK20 and CK 7 may be of helpful while searching for unknown primary in cases of metastatic deposit.

PD-L1 Expression in Colorectal Carcinoma: A Comparison of 3 Scoring Methods in a Cohort of Jordanian Patients.

Anti-programmed death-ligand 1 (PD-L1) treatments can improve colorectal carcinoma (CRC) survival; however, there is still controversy regarding the relationship between PD-L1 expression and the outcome of immunotherapeutic treatment and survival. The discrepancies are partly caused by the lack of a unified scoring system. This retrospective, cross-sectional study evaluated PD-L1 by immunohistochemistry in 127 CRC cases and compared the 3 scoring systems used to assess PD-L1: Tumor Percentage Score (TPS), Combined Positive Score (CPS), and immune cell (IC) score. Correlations were calculated using the χ 2 test. Kaplan-Meier curves with the Log-rank test were used to measure the contribution of PD-L1 expression to survival. PD-L1-positive rate were 29.9%, 57.5%, and 55.9% based on TPS, CPS, and IC score, respectively. TPS showed a better correlation with the clinicopathologic features being significantly higher with young age, T4, and adenocarcinomas (compared with mucinous/signet ring). TPS also showed an increasing trend with higher grade, lymph node stage, and male sex, although these variables were not significantly associated with PD-L1 expression. There was no correlation between PD-L1 expression and mismatch repair protein status in the 3 scoring methods. The probability of survival was higher for PD-L1-negative cases in the first 60 months after surgery if scored by the TPS method ( P =0.058). Future efforts correlating PD-L1 status with response to treatment are needed to decide on the best scoring method to be used for making therapy decisions.

PD-L1 and PD-L2 expression in colorectal cancer.

The programmed death-1 (PD-1) is an immune checkpoint molecule that suppresses T-cell response. The binding of PD-1 to PD-L1/PD-L2 results cytokine production, and T-cell proliferation are reduced. Tumors expressing PD-L1 and PD-L2 escape from cytotoxic T-cells and are exposed to tumor progression. For this reason, immunotherapy has become a new option in the treatment of cancer. In this study, we examined the PD-L1 and PD-L2 expression in colorectal carcinoma (CRC), and evaluated the relationship between clinicopathological parameters and CD8+ T cells. We evaluated CD8 expression in tumor-infiltrating lymphocytes and surrounding tumor lymphocytes with PD-L1, PD-L2 staining in tumor cells and immune cells formalin-fixed paraffin embedded samples of 124 patient diagnosed with CRC. Pearson Chi-Square, Fisher Exact Chi-Square, and Pearson Exact Chi-Square analyses were used in the analysis of the cross tables. Survival distributions predicted Kaplan--Meier method and it was evaluated using log-rank statistics. In our study, a significant correlation was found between PD-L1 expression and female sex and tumors with medullary morphology. No expression of PD-L2 was observed in tumors containing medullary morphology, and a statistically inverse relationship was observed between PD-L2 and the medullary component. PD-L1 positive tumor-infiltrating lymphocytes were determined to be an important predictor for recurrence-free survival. We believe that the evaluation of these parameters may be useful in the selection of patients who will benefit from immunotherapy in CRC cases.

Inverse relationship between Fusobacterium nucleatum amount and tumor CD274 (PD-L1) expression in colorectal carcinoma.

The CD274 (programmed cell death 1 ligand 1, PD-L1)/PDCD1 (programmed cell death 1, PD-1) immune checkpoint axis is known to regulate the antitumor immune response. Evidence also supports an immunosuppressive effect of Fusobacterium nucleatum. We hypothesised that tumor CD274 overexpression might be inversely associated with abundance of F. nucleatum in colorectal carcinoma. We assessed tumor CD274 expression by immunohistochemistry and F. nucleatum DNA within tumor tissue by quantitative PCR in 812 cases among 4465 incident rectal and colon cancer cases that had occurred in two prospective cohort studies. Multivariable logistic regression analyses with inverse probability weighting were used to adjust for selection bias because of tissue data availability and potential confounders including microsatellite instability status, CpG island methylator phenotype, LINE-1 methylation level and KRAS, BRAF and PIK3CA mutations. Fusobacterium nucleatum DNA was detected in tumor tissue in 109 (13%) cases. Tumor CD274 expression level was inversely associated with the amount of F. nucleatum in colorectal cancer tissue (P = 0.0077). For one category-unit increase in three ordinal F. nucleatum categories (negative vs. low vs. high), multivariable-adjusted odds ratios (with 95% confidence interval) of the low, intermediate and high CD274 categories (vs. negative) were 0.78 (0.41-1.51), 0.64 (0.32-1.28) and 0.50 (0.25-0.99), respectively (P trend = 0.032). Tumor CD274 expression level was inversely associated with the amount of F. nucleatum in colorectal cancer tissue, suggesting that different immunosuppressive mechanisms (i.e. PDCD1 immune checkpoint activation and tumor F.nucleatum enrichment) tend to be used by different tumor subgroups.

Clinical and Prognostic Significance of PD-L1, Beclin 1 and Fascin 1 Immunohistochemical Expression in Colorectal Carcinoma

Introduction: Colorectal Cancer (CRC) is a leading cause of death worldwide, and its incidence has been rising. It is important to identify novel markers that could predict the prognosis of the disease. Aim: To identify the impact of Programmed cell death-ligand 1 (PD-L1), Beclin 1, and Fascin 1 immunohistochemical expression on CRC behaviour and disease prognosis. Also, to assess the different surgical options (laparoscopic versus open) in the management of these patients. Materials and Methods: This cohort study included 72 cases of CRC, conducted in the Pathology, General Surgery, Internal Medicine, Clinical Oncology, and Medical Oncology Departments of the Faculty of Medicine, Zagazig University Hospitals, Egypt between June 2018 and June 2022. PD-L1, Beclin 1, and Fascin 1 expression were assessed immunohistochemically, and their prognostic significance were evaluated. Data were analysed using IBM Statistical Package for the Social Sciences (SPSS) 23.0 for Windows (SPSS Inc., Chicago, IL, USA) and NCSS 11 for Windows (NCSS LCC., Kaysville, UT, USA) in the Community Medicine Department, Faculty of Medicine, Zagazig University, Egypt. Results: A significant association was found between PDL1 expression and grading (p-value=0.02), lymphovascular invasion, distant metastasis (p-value=0.001), peritoneal spread (p-value=0.002), tumour budding (p-value=0.005), lymph node metastasis, and American Joint Committee on Cancer (AJCC) stage (p-value&lt;0.001). A significant association was also found between Beclin 1 expression and distant metastasis (p-value=0.001), lymphovascular invasion (p-value=0.004), lymphocytic infiltration (p-value=0.006), perineural invasion (p-value=0.01), peritoneal spread (p-value=0.002), and AJCC stage (p-value=0.007). A highly significant association between Fascin 1 expression and grading, lymphovascular invasion, tumour budding, lymph node metastasis, lymphocytic infiltration, perineural invasion, metastasis, peritoneal spread, and AJCC stage (p-value&lt;0.001) was detected. In patients with negative PD-L1 or high Fascin 1 expression, survival rates were lower. Beclin 1 expression was not associated with a favourable prognosis (p-value&gt;0.05). Conclusion: PD-L1, Beclin 1, and Fascin 1 expression are significantly elevated in CRC tissues and closely associated with adverse prognostic factors. PD-L1 and Fascin 1 expression have important prognostic value and may be of great help in identifying high-risk patients who will benefit the most from treatment.

PD-L1 Expression in Colorectal Adenocarcinoma Is Associated With the Tumor Immune Microenvironment and Epithelial-Mesenchymal Transition.

Colorectal carcinomas are the third-most common tumors in the world, and colorectal cancer ranks second in cancer-related deaths. Our aim in this study was to investigate the correlation between programmed cell death ligand 1 (PD-L1) expression and clinicopathologic parameters in colorectal carcinomas and their relationship to the tumor immune microenvironment, epithelial-mesenchymal transition (EMT), and microsatellite instability. We also investigated the predictive and prognostic role of PD-L1. One hundred patients with a diagnosis of colorectal adenocarcinoma who did not receive neoadjuvant therapy were included in the study. The relationships among the altered expression of PD-L1; vimentin; E-cadherin; mismatch repair status; and pathologic microenvironmental features, including the presence of tumor budding and CD8-positive tumor infiltrating lymphocytes (TILs), were assessed. Increased PD-L1 expression in tumor cells was associated with increased TILs (P = .013), high histologic grade (P = .011), advanced pathologic T stage (P = .007), lymph node metastasis (P = .002), distant metastasis (P < .001), perineural invasion (P = .009), high bud score (P = .023), EMT (P < .001), and shorter disease-free survival (P = .029). Overall, PD-L1 expression in colorectal carcinoma tumor cells is a marker of poor prognosis, and the positive correlation detected between EMT status and PD-L1 expression suggests that patients with the mesenchymal phenotype may be more likely to benefit from programmed cell death 1 protein/PD-L1 immunotherapy.

Hyal1 Expression in Colorectal Carcinoma Cell Migration and Invasiveness: Significance and Mechanism.

Objective To clarify the significance of hyaluronidsase 1 (Hyal1) expression in colorectal carcinoma (CRC) and its impact on tumor cell migration and invasiveness. Methods Human CRC cell lines SW480, HCT116, and SW620 were purchased, ELISA and western blot were used to detect the expression of Hyal1 in cells, CCK-8 assay to detect cell proliferation ability, cell scratch assay to check cell migration rate, and cell invasion was detected by the transwell assay. The correlation of Hyal1 with CRC cell migration and invasiveness capacities was analyzed. Result ELISA results showed that supernatant Hyal1 level was the lowest in SW480, highest in HCT116, with the level in SW620 in between (P < 0.05). No evident difference was identified by western blot in Hyal1 protein expression among the three cells (P > 0.05). The cell scratch assay and transwell assay showed that the migration and invasion ability of HCT116 cells was higher than that of SW620 (P < 0.05). In vitro, Hyal1 had a synergistic relationship with the invasiveness and migration capacities of CRC cells (P < 0.05). Conclusion Hyal1 is elevated in CRC and is consistent with the invasiveness and metastasis abilities of CRC cells. It is hoped that this research can provide reference for future prevention and treatment of CRC.

The clinicopathological features of programmed death ligand-1 expression in colorectal carcinoma

Few studies have addressed the clinicopathological features of colorectal cancer (CRC) that express programed death-ligand 1 (PD-L1). Various assays and scoring methodologies were used and thus inconsistent results were obtained. In this study, we aimed to investigate the relationship of PD-L1 expression in CRC with various clinicopathological variables using a standardized assay and scoring algorithm. Tissue microarrays were constructed from 91 random cases of CRC diagnosed at King Hussein Cancer Center (KHCC). Immunohistochemical (IHC) staining using the monoclonal antibody 22C3 was performed. Scoring using the standard "Combined Positive Score" (CPS) method was done. CPS of ≥1 was considered positive. Various clinicopathological features were collected from the medical records of the patients. Of the 91 cases, 49 (53.8%) were PD-L1 positive (CPS ≥1). PD-L1 expression was more frequent among moderately differentiated carcinomas (43 of 72 (59.7%) were positive compared to 6 of 19 (31.5%) poorly differentiated cases (P = 0.029)); among node negative cases (21 of 24 (87.5%) N0 cases were PD-L1 positive in contrast to 28 of 67 (41.8%) N1/N2 cases (P = <0.001)); among mucinous subtype (12 of 15 (80%) of cases (P = 0.02)); and among mismatch repair deficient (dMMR) (16 of 16 (100%) versus 11 of 30 (36.6%) MMR proficient (P = <0.001)). Age, gender, localization, and T or M stages were not significantly associated with PD-L1 expression. PD-L1 expression in CRC is associated with favorable prognostic features; namely, lower grade, N0, mucinous variant, and dMMR tumors.

Impact of Neoadjuvant Chemotherapy on SATB2 Expression in Colorectal Carcinomas: SATB2 Positivity is Preserved in Most Cases, but Down-Expressed in Effective Cases of Chemotherapy.

Special AT-rich sequence-binding protein 2 (SATB2) is a novel, diagnostically useful, and highly sensitive immunohistochemical marker for both primary and metastatic colorectal or appendiceal tumors. In the present study, we aimed to assess the impact of neoadjuvant chemotherapy on SATB2 expression in primary colorectal carcinomas and their corresponding liver metastases. Forty-four patients with colorectal carcinomas who received neoadjuvant chemotherapy were included. SATB2 expression in specimens of biopsy, resected primary colorectal carcinomas, and resected metastatic foci were examined by immunohistochemistry and compared to caudal-type homeobox transcription factor 2 (CDX2). Using a modified H-score, expressions were scored semiquantitatively for both staining intensity and tumor cell proportion with nuclear staining. SATB2 was positive in 43/44 cases (98%) in biopsy specimens, 42/44 cases (96%) in resected colorectal carcinomas with neoadjuvant chemotherapy, and 9/9 cases (100%) with liver metastases. However, these expressions were variably decreased, and the H-score was lower in resected colorectal carcinomas (158 ± 69) than in biopsy specimens (174 ± 60) (p < 0.01). The proportion of SATB2-positive area of colorectal carcinoma was 93% in metastatic foci, while the CDX2-positive area was 78%. When categorized by histopathological tumor regression, the most effective tumors of chemotherapy showed the lowest H-score in resected colorectal carcinomas among the three groups (p < 0.01). SATB2 is a useful marker for both primary colorectal carcinoma and corresponding liver metastases, even with neoadjuvant chemotherapy. However, caution should be exercised when performing needle biopsy for metastatic foci with neoadjuvant therapy because expressions could be decreased, especially in chemotherapy-effective cases, and show immunohistochemically negative results.

Human Epidermal Growth Factor Receptor 2 (HER2) Expression in Colorectal Carcinoma: A Potential Area of Focus for Future Diagnostics.

ObjectiveIn this study, we aimed to explore the potential diagnostic utility of human epidermal growth factor receptor 2 (HER2) expression in colorectal carcinoma. We investigated the association of HER2 expression with the type and grade of the tumor along with the pattern, staining intensity, and the percentage of cells stained.MethodsThis was an observational study involving 50 cases of colorectal carcinoma that underwent immunohistochemistry to analyze the HER2 expression.ResultsThe positive expression of HER2 was seen in 16 (32%) cases. The majority of the study population was between the fifth-seventh decades of life. The most commonly diagnosed tumor was conventional adenocarcinoma with grade II, cytoplasmic pattern, +2 positivity, and moderate intensity. The maximum positivity for HER2 was seen in tumors of the rectum in eight (16%) cases.ConclusionA substantial rate of HER2 overexpression paves the way for it to become a potential future target in cancer therapeutics.

Diagnostic value of IMP3 expression in colorectal carcinoma and adenoma

Diagnostic value of IMP3 expression in colorectal carcinoma and adenoma

Immunohistochemical expression of CD8+ tumor-infiltrating lymphocytes and Foxp3 expression in colorectal carcinoma

Background/aim Colorectal cancer (CRC) is third among diagnosed tumors (6.1%) and second according to mortality (9.2%). Disease prognosis is determined not only by the histologic and molecular features of the tumor but also by the host response. Histologic distributions of tumor-infiltrating lymphocytes (TIL) in the microenvironment can be correlated with staging and prognosis of CRC patients. Abundance of CD8+ T lymphocytes has been associated with good prognosis in different types of solid tumors. The association between tumor cell expression of FoxP3 and tumor infiltration by FoxP3-expressing T lymphocytes with prognosis is still controversial. The aim of this study is to evaluate CD8+ TILs and expression of FoxP3 in CRC and to correlate their expression with patients’ clinicopathological parameters.Materials and methods Tumor paraffin blocks and clinicopathological data of 60 patients with CRC were obtained from the Pathology Department at Cairo University. The density of CD8 TILs and FoxP3 expression was assessed immunohistochemically and evaluated by image analysis in CRC specimens using area percentage parameter. The CD8+ cell tumor infiltrate and FoxP3 expression were classified into scanty, moderate, and abundant.Results CD8+ TIL in the present study was insignificantly correlated with the clinicopathological parameters, and no correlation was detected between FoxP3 and CD8 expression (P>0.05). However, FOXP3 expression was significantly correlated with tumor grade, nodal status, distant metastasis, tumor stage, and Dukes’ classification (P<0.01).Conclusion The presence of FoxP3 expression in CRC correlates with favorable pathological prognostic parameters. Cancer colon progression is influenced by host immune response. More studies are needed to assess the role of tumor microenvironment in CRC prognosis.

Loss of SATB2 Occurs More Frequently Than CDX2 Loss in Colorectal Carcinoma and Identifies Particularly Aggressive Cancers in High-Risk Subgroups

Simple SummaryThe immunohistochemical analysis of Special AT-rich sequence-binding protein 2 (SATB2) is increasingly being used to detect colorectal differentiation. Our study aimed to investigate SATB2 expression levels and the prognostic relevance of SATB2 loss in colorectal carcinoma (CRC), especially in comparison with CDX2, the standard marker of colorectal differentiation. We tested SATB2 expression in 1039 CRCs and identified SATB2 as a strong prognosticator in the overall cohort as well as in specific subcohorts, including high-risk subgroups. Compared to CDX2, SATB2 showed a higher prognostic power but was lost at a much higher frequency, generally rendering SATB2 as the less sensitive marker for colorectal differentiation compared to CDX2.Background: Special AT-rich sequence-binding protein 2 (SATB2) has emerged as an alternative immunohistochemical marker to CDX2 for colorectal differentiation. However, the distribution and prognostic relevance of SATB2 expression in colorectal carcinoma (CRC) have to be further elucidated. Methods: SATB2 expression was analysed in 1039 CRCs and correlated with clinicopathological and morphological factors, CDX2 expression as well as survival parameters within the overall cohort and in clinicopathological subgroups. Results: SATB2 loss was a strong prognosticator in univariate analyses of the overall cohort (p < 0.001 for all survival comparisons) and in numerous subcohorts including high-risk scenarios (UICC stage III/high tumour budding). SATB2 retained its prognostic relevance in multivariate analyses of these high-risk scenarios (e.g., UICC stage III: DSS: p = 0.007, HR: 1.95), but not in the overall cohort (DSS: p = 0.1, HR: 1.25). SATB2 loss was more frequent than CDX2 loss (22.2% vs. 10.2%, p < 0.001) and of higher prognostic relevance with only moderate overlap between SATB2/CDX2 expression groups. Conclusions: SATB2 loss is able to identify especially aggressive CRCs in high-risk subgroups. While SATB2 is the prognostically superior immunohistochemical parameter compared to CDX2 in univariate analyses, it appears to be the less sensitive marker for colorectal differentiation as it is lost more frequently.

High-throughput virtual screening and preclinical analysis identifies CB-1, a novel potent dual B-Raf/c-Raf inhibitor, effective against wild and mutant variants of B-Raf expression in colorectal carcinoma.

Paradoxical Raf activation via Raf dimerization is a major drawback of wild/mutant B-Raf inhibitors. Herein, we report that CB-1 a novel, potent B-Raf/c-Raf dual inhibitor, effective against colon cancer cells, irrespective of their genetic status. High-throughput virtual screening of the ChemBridge library against wild B-Raf (B-RafWT), mutant B-Raf (B-RafV600E), and c-Raf was performed using an automated protocol with the AutoDock-VINA. Caco-2 and HT-29 cells were used. Of the 23,365 compounds screened computationally, CB-1 showed the highest binding energy towards B-RafWT with a ΔGbinding score of -13.0kcal/mol. The compound was also predicted to be effective against B-RafV600E and c-Raf molecules with ΔGbinding energies of -10.6 and -10.1kcal/mol, respectively. The compound inhibited B-RafWT, B-RafV600E and c-Raf kinases with IC50 values of 27.13, 51.70, and 40.23 nM, respectively. The GI50 value of CB-1 was 247.9 nM in B-RafWT-expressing Caco-2 cells and 352.4 nM in B-RafV600E-expressing HT-29 cells. Dose-dependent increases in total apoptosis and G1 cell cycle phase arrest was observed in CB-1-treated colon cancer cells. The compound decreased B-Raf expression in both wild and mutant colon cancer cells. CB-1, a novel, potent dual B-Raf/c-Raf inhibitor was effective against colon cancer cells bearing wild-type and mutant variants of B-Raf expression.

When Activator and Inhibitor of PPARα Do the Same: Consequence for Differentiation of Human Intestinal Cells.

Peroxisome proliferator-activated receptor α (PPARα) is a ligand-dependent transcription factor that plays a role in various processes including differentiation of several cell types. We investigated the role of PPARα in the differentiation of intestinal cells using HT-29 and Caco2 cell lines as a model as well as human normal colon and colorectal carcinoma tissues. We detected a significant increase in PPARα expression in differentiated HT-29 cells as well as in normal surface colon epithelium where differentiated cells are localised. Thus, it seems that PPARα may play a role in differentiation of intestinal cells. Interestingly, we found that both PPARα activators (fenofibrate and WY-14643) as well as its inhibitor (GW6471) regulated proliferation and differentiation of HT-29 cells in vitro in the same way. Both compounds led to a decrease in proliferation accompanied by a significant increase in expression of villin, intestinal alkaline phosphatase (differentiation markers). Moreover, the same trend in villin expression was observed in Caco2 cells. Furthermore, villin expression was independent of subcellular localisation of PPARα. In addition, we found similar levels of PPARα expression in colorectal carcinomas in comparison to adjacent normal epithelium. All these findings support the hypothesis that differentiation of intestinal epithelium is PPARα-independent.