Dukes Stage Research Articles

Global Distribution of Colorectal Cancer Staging at Diagnosis: An Evidence Synthesis.

Stage at diagnosis is a key prognostic factor for colorectal cancer (CRC) survival. We aimed to assess the global distribution of CRC staging at diagnosis using population-based or hospital-based registry data. We systematically searched in MEDLINE and Embase from their inception until 6 December 2023. Grey literature was searched through published cancer reports. Studies from population-based or hospital-based cancer registries reporting the stage distribution of diagnosed CRC were included. We extracted stage-specific proportions among patients with CRC based on TNM, SEER, or Dukes staging systems. Subgroup analyses by sex, age, tumor site, calendar period, and status of population-based screening were performed. Correlations between the Human Development Index (HDI), Socio-Demographic Index (SDI), and the distant metastatic CRC proportion were also evaluated. 84 eligible studies/reports from 46 countries were analyzed, covering 3.8 million patients. Among 36 countries included in the main analysis, the most recent distant metastatic CRC proportions varied from 16.2% in Puerto Rico to 28.2% in Oman and Latvia, with a median (interquartile range) of 23.7% (21.8%-26.3%). Higher metastatic proportions were observed in younger patients, those with colon cancer, and those without screening implementation, with no apparent difference between males and females. Over time, some countries/regions, such as Southern Portugal (36.5% in 2000 to 22.2% in 2016), showed declining proportions of metastatic CRC, while others remained stable (e.g., Austria, Belgium) or increased (e.g., United States, Slovenia, Spain). Higher HDI and SDI were correlated with lower proportions of distant metastatic CRC (HDI: ρ=-0.48; SDI: ρ=-0.26). Global disparities in CRC staging exist, indicating a need for targeted interventions to enhance early detection and management, especially in high-metastasis areas.

Combined High-Throughput Proteomics and Random Forest Machine-Learning Approach Differentiates and Classifies Metabolic, Immune, Signaling and ECM Intra-Tumor Heterogeneity of Colorectal Cancer.

Colorectal cancer (CRC) is a frequent, worldwide tumor described for its huge complexity, including inter-/intra-heterogeneity and tumor microenvironment (TME) variability. Intra-tumor heterogeneity and its connections with metabolic reprogramming and epithelial-mesenchymal transition (EMT) were investigated with explorative shotgun proteomics complemented by a Random Forest (RF) machine-learning approach. Deep and superficial tumor regions and distant-site non-tumor samples from the same patients (n = 16) were analyzed. Among the 2009 proteins analyzed, 91 proteins, including 23 novel potential CRC hallmarks, showed significant quantitative changes. In addition, a 98.4% accurate classification of the three analyzed tissues was obtained by RF using a set of 21 proteins. Subunit E1 of 2-oxoglutarate dehydrogenase (OGDH-E1) was the best classifying factor for the superficial tumor region, while sorting nexin-18 and coatomer-beta protein (beta-COP), implicated in protein trafficking, classified the deep region. Down- and up-regulations of metabolic checkpoints involved different proteins in superficial and deep tumors. Analogously to immune checkpoints affecting the TME, cytoskeleton and extracellular matrix (ECM) dynamics were crucial for EMT. Galectin-3, basigin, S100A9, and fibronectin involved in TME-CRC-ECM crosstalk were found to be differently variated in both tumor regions. Different metabolic strategies appeared to be adopted by the two CRC regions to uncouple the Krebs cycle and cytosolic glucose metabolism, promote lipogenesis, promote amino acid synthesis, down-regulate bioenergetics in mitochondria, and up-regulate oxidative stress. Finally, correlations with the Dukes stage and budding supported the finding of novel potential CRC hallmarks and therapeutic targets.

Pokemon inhibits Bim transcription to promote the proliferation, anti-anoikis, invasion, histological grade, and dukes stage of colorectal neoplasms

PurposeThis study aims to determine whether Pokemon regulates Bim activity in colorectal carcinoma (CRC) carcinogenesis.MethodsClinical tissue samples were analyzed to detect the expression and clinicopathological significance of Pokemon and Bim in CRC. Proliferation, apoptosis, and invasion assays were conducted to identify the regulatory effect of Pokemon on Bim. The combined treatment effects of Pokemon knockdown and diamminedichloroplatinum (DDP) were also examined.ResultsImmunohistochemical analysis of 80 samples of colorectal epithelia (CRE), 80 cases of colorectal adenoma (CRA), and 160 of CRC samples revealed protein expression rates of 23.8%, 38.8%, and 70.6% for Pokemon, and 88.8%, 73.8%, and 31.9% for Bim, respectively. A significant negative correlation was observed between Pokemon and Bim expression across the CRE, CRA, and CRC lesion stages. In CRC, higher Pokemon and lower Bim expression correlated with higher histological grades, advanced Dukes stages, and increased cancer invasion. In both LoVo and HCT116 cells, overexpression of Pokemon significantly reduced Bim expression, leading to increased proliferation, resistance to anoikis, and cell invasion. Additionally, Pokemon overexpression significantly decreased DDP-induced Bim expression, reduction of anti-apoptosis and invasion, whereas Pokemon knockdown resulted in the opposite effects.ConclusionThese findings suggest that Pokemon inhibits Bim transcription, thereby promoting CRC proliferation, resistance to apoptosis, invasion, and advancing histological grade and Dukes staging. Pokemon knockdown enhances the therapeutic efficacy of DDP in the treatment of CRC.

P4HA3 promotes colon cancer cell escape from macrophage phagocytosis by increasing phagocytosis immune checkpoint CD47 expression.

Cancer immunotherapies have greatly changed the prospects for the therapy of many malignancies, including colon cancer. Macrophages as the effectors of cancer immunotherapy provide considerable promise for cancer treatment. Prolyl 4-hydroxylase subunit alpha 3 (P4HA3) plays a cancer-promoting role in a variety of cancers, including colon cancer. In the present work, we provided evidence for the first time that P4HA3 promoted colon cancer cell escape from macrophage phagocytosis, and preliminarily explored its possible molecular mechanism. Immunohistochemistry was used to detect the expression of P4HA3 in tissues. Bioinformatics methods were used to analyze the tumor public databases (including TCGA database and GEO database). Macrophage phagocytosis assay and flow cytometric analysis were used to detect the phagocytic capacity of macrophages. Western blot and qRT-PCR were used to detect the expression of related markers (such as P4HA3, CD47, CD24, IL-34, and M-CSF). First, we found that P4HA3 was significantly and highly expressed in both colon cancer tissues and cells, and that P4HA3 had a positive correlation with lymph node metastasis, Dukes stage and also strongly correlated with poorer survival. Subsequently, we found that P4HA3 was strongly associated with the macrophage infiltration level in colon cancer. Immediately we also found that decreasing P4HA3 expression promoted macrophage phagocytosis in colon cancer cells, whereas P4HA3 overexpression produced the opposite effect. Finally, we demonstrated that P4HA3 promoted the expression of cluster of differentiation 47 (CD47) in colon cancer cells. Moreover, P4HA3 caused colon cancer cells to secrete Interleukin 34 (IL34) and Macrophage colony stimulating factor (M-CSF), which further induced macrophages to differentiate to M2 type and thereby contributed to the progression of colon cancer. We have demonstrated that P4HA3-driven CD47 overexpression may act as an escape mechanism, causing colon cancer cells to evade phagocytosis from macrophages.

Clinicopathological Profile of Colorectal Adenocarcinoma in the Anatomical Pathology Laboratory of Dr. Soedarso Hospital Pontianak

Background: Colorectal adenocarcinoma is ranked third globally in the category of the most commonly diagnosed cancers, as reported by the Global Burden of Cancer (GLOBOCAN) survey in 2020. This condition ranks as the second leading cause of death from cancer with a mortality of 9.40%. Therefore, this study aimed to discover the clinicopathological profile of colorectal adenocarcinoma in the Anatomical Pathology Laboratory of Dr. Soedarso Hospital Pontianak in 2017–2020. Methods: This descriptive study used secondary data from the histopathology reports of the Anatomical Pathology Laboratory obtained by total sampling, which were processed using Microsoft Excel. Results: The total number of colorectal adenocarcinoma patients was 148, and the majority were in the category of 53–61 years, accounting for 30.41%, and 51.35% were female. The majority of the anatomical sites of tumors were in the “Rectum Not Otherwise Specified (NOS)”, with a percentage of 35.14%. The most frequent histopathological subtype was adenocarcinoma NOS, accounting for 85.81%, while the majority of differentiation grade was well differentiated, with a percentage of 90.54%. The most TNM (Tumor, Node, Metastatic) staging system of the American Joint Committee on Cancer (AJCC) was IIA (33.78%). Based on the classification using the Dukes staging system, the most stage suffered by patients was C2 with a percentage of 39.86%. Conclusions: In conclusion, the incidence of diagnosed colorectal adenocarcinoma cases increased yearly in the Anatomical Pathology Laboratory of Dr. Soedarso Hospital Pontianak. Further study of the clinical characteristics and analysis of risk factors in colorectal adenocarcinoma patients were important to support the development of effective preventive strategies.

Evaluating the Efficacy of Fecal Occult Blood Test and Tumor Marker Combined Screening for Colorectal Cancer

Objective: To analyze the screening effectiveness of combining the fecal occult blood test with tumor marker detection for colorectal cancer. Methods: A total of thirty patients with colorectal cancer and thirty patients with benign colon hyperplasia who received treatment from January 2020 to January 2023 were selected. These patients were assigned to the observation group and the control group, respectively. All patients in both groups underwent both fecal occult blood tests and tumor marker detection. The levels of tumor markers between the two groups were compared, the tumor marker levels in different stages were assessed within the observation group, and the positive detection rates for single detection and combined detection were compared. Results: The levels of various tumor markers in the observation group were significantly higher than those in the control group (P < 0.05). Furthermore, as the Duke stage increased within the observation group, the levels of various tumor markers also increased (P < 0.05). The positive detection rate of the combined test was notably higher than that of single detection (P < 0.05). Conclusion: Combining the fecal occult blood test with tumor marker detection in colorectal cancer screening can significantly improve the overall detection rate.

Biliary Carcinoembryonic Antigen (CEA) Levels: The Role in Detection of Occult Hepatic Metastases in Colorectal Carcinoma

Objective: Colorectal carcinoma (CRC) is a major public health concern, often complicated by hepatic metastasis. Despite the widespread use of serum Carcinoembryonic Antigen (CEA) for postoperative monitoring, early detection of hepatic metastasis remains elusive. This study aims to evaluate the prognostic significance of CEA levels in both peripheral blood and gallbladder bile for predicting hepatic metastasis in CRC patients, and to explore its potential utility for personalized treatment regimens. Materials and Methods: A prospective randomized study was conducted over three years, enrolling 31 participants, 21 of whom underwent curative operations for CRC. Preoperative and postoperative CEA levels were assessed with rigorous diagnostic imaging and histological examinations. Patients were stratified into groups based on liver metastasis, postoperative chemotherapy, and CEA levels in both blood and bile. Results: A statistically significant difference was found in biliary CEA levels between patients with (mean = 129.22 ng/ml) and without intraoperative liver metastasis (mean = 33.16 ng/ml), p-value = 0.01. Serum and biliary CEA levels were robustly correlated, p > 0.05. Biliary CEA values differed significantly across Duke's stages, with particular significance between the control group and Duke's stage D (p < 0.001). Among patients without intraoperative liver metastasis but with biliary CEA >10 ng/ml, 80% developed liver metastasis postoperatively despite undergoing chemotherapy. Conclusion: This research highlights the pivotal role of biliary CEA levels in diagnosing latent hepatic metastases in colorectal cancer. Demonstrating superior sensitivity compared to traditional serum tests, biliary CEA presents a potential shift in early detection. Elevated levels suggest expanded therapeutic options, from timely hepatectomies to adopting radiofrequency ablation and anti-CEA monoclonal antibodies, enhancing the likelihood of better patient outcomes. Essentially, these findings enhance our understanding of colorectal cancer, emphasizing the importance of personalized care, early diagnosis, and customized treatments, thereby directing us toward better patient outcomes.

Differential expression of PLAC1 and Netrin-1 in liver metastasis of colorectal cancer and its predictive value

ObjectiveTo explore the differential expression of placental specific gene 1 (PLAC1) and neurite guidance factor 1 (netrin-1) in colorectal cancer (CRC) liver metastasis and its predictive value.MethodsParaffin specimens of primary CRC were selected, including 60 simple colorectal cancer specimens and 47 liver metastasis specimens. At the same time, 40 cases of normal colorectal mucosa were taken as the control group. The expression of PLAC1 and Netrin-1 in tissue was detected by immunohistochemistry (IHC). The correlation between PLAC1 and Netrin-1 expression and clinicopathological characteristics of patients with CRC liver metastases was analyzed. Logistic analysis was adopted to analyze the influencing factors of liver metastasis in CRC. A prediction model was established and ROC curve was used to detect the discrimination of the prediction model. The clinical value of PLAC1 and netrin-1 in predicting liver metastasis of CRC was analyzed using ROC curve. The relationship between the expression of PLAC1 and netrin-1 and the prognosis of CRC patients with liver metastasis was analyzed using Kaplan Meier survival curve.ResultsThe positive staining of PLAC1 and netrin-1 was mainly located in the cytoplasm by IHC detection. Positive expression of PLAC1 and netrin-1 in CRC tissues was markedly higher than that in normal colorectal mucosal epithelium (P < 0.05). Positive expression of PLAC1 in metastatic group was higher than that in non-metastatic group without significant difference (P > 0.05). The metastasis group had much higher positive expression of netrin-1 than the non-metastasis group (P < 0.05). The content of PLAC1 in the tissues of CRC with liver metastasis had a close relationship with differentiation degree and lymph node metastasis (P < 0.05). The expression of Netrin-1 in the tissues of CRC with liver metastasis was associated with Dukes stage, differentiation degree and lymph node metastasis (P < 0.05). Logistic regression analysis showed that Dukes stage, differentiation, lymph node metastasis, CEA, Alb and D-dimer were the independent risk factors for liver metastasis of CRC (P < 0.05). The model was constructed according to the regression coefficients and constant terms, and the discrimination of the prediction model was evaluated using ROC curve, with the AUC of 0.903 (95% CI 0.831 ~ 0.975), the sensitivity of 93.80%, the specificity of 80.00%, and the Jordan index of 0.738. The AUC of PLAC1 and netrin-1 alone and combined detection to predict liver metastasis of CRC were 0.805, 0.793 and 0.921, respectively. The survival time of patients with positive PLAC1 and netrin-1 expression were sharply shorter than that of the patients with negative expression (P < 0.05).ConclusionsThe expression of PLAC1 and netrin-1 was strongly increased in CRC with liver metastasis, which had a certain clinical value in predicting liver metastasis of CRC. Dukes stage, differentiation degree, lymph node metastasis, CEA, Alb and D-dimer were independent risk factors for liver metastasis of CRC, and the model based on these indicators had good discrimination for effectively evaluating the risk of liver metastasis in CRC.

Intussusception revealing right colonic adenocarcinoma in a 61-year-old woman: a case report

IntroductionAdult Intussusception is an uncommon diagnosis, with one to three cases occurring in a population of 1,000,000 per year, primarily due to underlying pathological lead points, of which 70% are malignant. Lipoma is the most common benign tumour, and primary adenocarcinoma is the most common malignant one. Early diagnosis and treatment are essential to reducing poor outcomes, including ischemia, perforation, and sepsis. Computed tomography imaging is a modality of choice for diagnosis. With a diagnostic accuracy of up to 100% and a specificity of up to 71%. Surgical intervention is the definitive treatment, and the decision is taken according to the situation.Case presentationA 61-year-old Black African female presented to our surgical casualty with intermittent colicky abdominal pain for 1 month. After that, she started to experience abdominal swelling in the right iliac fossa. A CT scan confirmed the presence of colo-colic Intussusception, cecum-ascending-transverse colon. Laparotomy was scheduled, and a right hemicolectomy was done accordingly. Diagnosis of adenocarcinoma (Dukes stage B2) was made histologically.ConclusionIntussusception in adults is a challenging diagnosis requiring high clinical suspicion and has a high incidence of fatal complications. Colonic cancer can be worsening by any infection and chronic medical problem. CT imaging is the lifesaving modality of choice for diagnosis. Good patient outcomes depend on timely diagnosis and recruitment of a multi-disciplinary team.

Prognostic Value of BRAF, Programmed Cell Death 1 (PD1), and PD Ligand 1 (PDL1) Protein Expression in Colon Adenocarcinoma.

Patients with colorectal cancer in different stages show variable outcomes/therapeutic responses due to their distinct tumoral biomarkers and biological features. In this sense, this study aimed to explore the prognostic utility of BRAF, programmed death-1 (PD1), and its ligand (PDL1) protein signatures in colon adenocarcinoma. The selected protein markers were explored in 64 archived primary colon adenocarcinomas in relation to clinicopathological features. BRAF overexpression was found in 39% of the cases and was significantly associated with grade 3, N1, advanced Dukes stage, presence of relapse, and shorter overall survival (OS). PD1 expression in the infiltrating immune cells (IICs) exhibited significant association with T2/T3, N0/M0, early Dukes stage, and absence of relapse. PDL1 expression in IICs is significantly associated with advanced nodal stage/distant metastasis, advanced Dukes stage, and shorter OS. Meanwhile, PDL1 expression in neoplastic cells (NC) was associated with the advanced lymph node/Dukes stage. A positive combined expression pattern of PDL1 in NC/IICs was associated with poor prognostic indices. Tumor PDL1 expression can be an independent predictor of OS and DFS. The multivariate analyses revealed that short OS was independently associated with the RT side location of the tumor, PD1 expression in stromal IICs, and PDL1 expression in NC. In conclusion, overexpression of BRAF in colon adenocarcinoma is considered a poor prognostic pathological marker. In addition, PDL1 expression in NC is considered an independent prognostic factor for DFS/OS. Combined immunohistochemical assessment for BRAF and PD1/PDL1 protein expressions in colon adenocarcinoma might be beneficial for selecting patients for future targeted therapy.

A clinicopathological study of the correlation of level of carcinoembryonic antigen in colorectal carcinoma in a tertiary care medical college in Eastern India

Background: For different cancers, different tumor markers are overexpressed. Carcinoembryonic antigen (CEA) levels are raised commonly in adenocarcinoma of colon, breast, and lung. About 80% of primary colorectal cancers (CRCs) show high serum CEA levels. Hence, CEA levels can be used as a prognostic factor - for both metastasis and recurrence - with persistent high levels. Currently, the American Society of Clinical Oncology recommends CEA as tumor marker and prognostic indicator in CRC patients. Aim and Objectives: The aim of the study was to determine the high level of CEA in CRCs and to study the level of CEA in cases of metastases and recurrence and to determine the sensitivity of CEA value in prognosis of CRCs. Materials and Methods: It was a prospective observational study. All the patients more than 20 years of age and of both sexes were included in this study who were diagnosed with primary localized CRC, metastatic CRC, or recurrent disease. The study was performed in the Department of General Surgery, R. G. Kar Medical College and Hospital between January 2020 and August 2021. Results: Total cases were classified according to Duke staging - 10% in Group A, 19% in Group B, 38% in Group C, and 33% in Group D. In Group A and B, about 35% of patients had increased CEA levels, whereas in Group C and D, more than 90% of patients had increased CEA levels with significant P &lt; 0.05. Overall 80% of the patients had elevated CEA levels, so remaining 20% had normal values. In cases of recurrent colorectal carcinoma, &gt;90% of patients had raised CEA levels, and in metastatic disease cases, CEA level was significantly raised in 82% of patients. Conclusion: In case of colorectal CA, preoperative raised serum CEA level is associated with poor prognosis, as chance of recurrence increased. Along with that increased CEA level is also associated with increased chance of metastatic disease. Hence, serial monitoring of serum CEA level in postoperative period may help to detect any recurrence early, thus reducing chance of mortality and morbidity.

LINC01285 promotes proliferation and metastasis of colorectal cancer cells by regulating epithelial-mesenchymal transition

To clarify the mechanism by which LINC01285 regulates proliferation and migration of colorectal cancer (CRC) cells and the clinical implications. We analyzed the expression of LINC01285 in CRC tissues and normal tissues using data from Starbase public database. We also examined the expression levels of LINC01285 in 70 pairs of CRC and adjacent tissue samples collected from our center and in different CRC cell lines using RT-qPCR, and analyzed the correlation of LINC01285 expression with the clinicopathological parameters and tumor-free survival time of the patients. In CRC cell lines (SW620 and HT-29), the changes in cell proliferation, apoptosis, metastasis and epithelial-mesenchymal transition (EMT) phenotype following LINC01285 knockdown were analyzed using CCK-8 assay, flow cytometry, Transwell assay and Western blotting. The TCGA-COAD transcriptome sequencing data obtained from the Starbasev3.0 public database revealed a significantly higher expression level of LINC01285 in CRC tissues than in adjacent tissues (P=0.00016), which was verified by RT-qPCR results of the clinical samples (P=0.0002). In CRC patients, the expression level of LINC01285 was closely correlated with histological differentiation of the tumor (P=0.036), T classification (P=0.000), lymph node metastasis (P=0.001), TNM stage (P=0.000), Duke stage (P=0.009) and relapse-free survival (P=0.0102). In SW620 and HT-29 cells, which expressed significantly higher levels of LINC01285 than normal colorectal mucosal cells (P < 0.001), LINC01285 knockdown significantly inhibited cell proliferation (P < 0.001), increased early apoptosis, late apoptosis and total apoptosis rates (P < 0.05), suppressed cell migration and invasion (P < 0.001), upregulated the expression of E-cadherin (P < 0.001), and downregulated the expression of N-cadherin (P < 0.001). The expression level of LINC01285, which modulates the EMT pathway to regulate the proliferation, apoptosis and metastasis of CRC cells, is closely correlated with the prognosis of CRC patients.

Trends in the full blood count blood test and colorectal cancer detection: a longitudinal, case-control study of UK primary care patient data [version 2; peer review: 2 approved, 1 not approved

The full blood count (FBC) is a common blood test performed in general practice. It consists of many individual parameters that may change over time due to colorectal cancer. Such changes are likely missed in practice. We identified trends in these FBC parameters to facilitate early detection of colorectal cancer. We performed a retrospective, case-control, longitudinal analysis of UK primary care patient data. LOWESS smoothing and mixed effects models were derived to compare trends in each FBC parameter between patients diagnosed and not diagnosed over a prior 10-year period. There were 399,405 males (2.3%, n = 9,255 diagnosed) and 540,544 females (1.5%, n = 8,153 diagnosed) in the study. There was no difference between cases and controls in FBC trends between 10 and four years before diagnosis. Within four years of diagnosis, trends in many FBC levels statistically significantly differed between cases and controls, including red blood cell count, haemoglobin, white blood cell count, and platelets (interaction between time and colorectal cancer presence: p <0.05). FBC trends were similar between Duke's Stage A and D colorectal tumours, but started around one year earlier in Stage D diagnoses. Trends in FBC parameters are different between patients with and without colorectal cancer for up to four years prior to diagnosis. Such trends could help earlier identification.

Benefit of Biennial Fecal Occult Blood Screening on Colorectal Cancer in England: A Population-Based Case-Control Study.

BackgroundThe English national bowel cancer screening program offering a guaiac fecal occult blood test began in July 2006. In randomized controlled trials of guaiac fecal occult blood test screening, reductions in mortality were accompanied by reductions in advanced stage colorectal cancer (CRC). We aimed to evaluate the effect of participation in the national bowel cancer screening program on stage-specific CRC incidence as a likely precursor of a mortality effect.MethodsIn this population-based case-control study, cases were individuals diagnosed with CRC aged 60-79 years between January 1, 2012, and December 31, 2013. Two controls per case were matched on geographic region, gender, date of birth, and year of first screening invitation. Screening histories were extracted from the screening database. Conditional logistic regression with correction for self-selection bias was used to estimate odds ratios (odds ratios corrected for self-selection bias [cOR]) and 95% confidence intervals (CIs) by Duke stage, sex, and age.Results14 636 individuals with CRC and 29 036 without were eligible for analysis. The odds of CRC (any stage) were increased within 30 days of a screening test and decreased thereafter. No reduction in CRC (any stage) among screened individuals compared with those not screened was observed (cOR = 1.00, 95% CI = 0.89 to 1.15). However, screened individuals had lower odds of Duke stage D CRC (cOR = 0.68, 95% CI = 0.50 to 0.93). We estimate 435 fewer Duke D CRC by age 80 years in 100 000 people screened biennially between ages 60 and 74 years compared with an unscreened cohort.ConclusionThe impact of colorectal screening on advanced CRC incidence suggests that the program will meet its aim of reducing mortality.

SRSF9 promotes colorectal cancer progression via stabilizing DSN1 mRNA in an m6A-related manner

BackgroundSerine/arginine-rich splicing factor 9 (SRSF9) is a classical RNA-binding protein that is essential for regulating gene expression programs through its interaction with target RNA. Whether SRSF9 plays an essential role in colorectal cancer (CRC) progression and can serve as a therapeutic target is largely unknown. Here, we highlight new findings on the role of SRSF9 in CRC progression and elucidate the underlying mechanism.MethodsCRC cell lines and clinical tissue samples were used. qRT-PCR, Western blotting, immunohistochemistry (IHC), gain- and loss-of-function assays, animal xenograft model studies, bioinformatic analysis, methylated single-stranded RNA affinity assays, gene-specific m6A quantitative qRT-PCR, dual-luciferase reporter assays and RNA stability assays were performed in this study.ResultsThe expression level of SRSF9 was higher in CRC cell lines than that in an immortal human intestinal epithelial cell line. Overexpression of SRSF9 was positively associated with lymph node metastasis and Dukes stage. Functionally, SRSF9 promoted cell proliferation, migration and invasion in vitro and xenograft growth. The results of bioinformatic analysis indicated that DSN1 was the downstream target of SRSF9. In CRC cells and clinical tissue samples, the expression of SRSF9 was positively associated with the expression of DSN1. Knockdown of DSN1 partially inhibited the SRSF9-induced phenotype in CRC cells. Mechanistically, we further found that SRSF9 is an m6A-binding protein and that m6A modifications were enriched in DSN1 mRNA in CRC cells. Two m6A modification sites (chr20:36773619–36773620 and chr20:36773645–chr20:36773646) in the SRSF9-binding region (chr20:36773597–36773736) of DSN1 mRNA were identified. SRSF9 binds to DSN1 in an m6A motif- and dose-dependent manner. SRSF9 modulates the expression of DSN1 in CRC cells. Such expression regulation was largely impaired upon methyltransferase METTL3 knockdown. Moreover, knockdown of SRSF9 accelerated DSN1 mRNA turnover, while overexpression of SRSF9 stabilized DSN1 mRNA in CRC cells. Such stabilizing was also weakened upon METTL3 knockdown.ConclusionOverexpression of SRSF9 was associated with lymph node metastasis and Dukes stage in CRC. Knockdown of DSN1 eliminated the effects by SRSF9 overexpression in CRC. Our results indicated that SRSF9 functions as an m6A-binding protein (termed “reader”) by enhancing the stability of DSN1 mRNA in m6A-related manner. Our study is the first to report that SRSF9-mediated m6A recognition has a crucial role in CRC progression, and highlights SRSF9 as a potential therapeutic target for CRC management.

A CLINICOPATHOLOGICAL STUDY OF THE CORRELATION OF CARCINOEMBRYONIC ANTIGEN IN COLORECTAL CARCINOMA: IN A TERTIARY CARE MEDICAL COLLEGE SET-UP IN EASTERN INDIA

Introduction: Carcino embryonic antigen(CEA) is overexpressed primarily by adenocarcinomas including colon, rectum, breast and lung. It should be noted that more than 90% of primary colorectal carcinomas produce CEA.CEA is a classic tumour marker for CRC, which is used to monitor the recurrence of the disease and as a prognostic factor for the patients. In the present scenario, the serum CEAtest is recommended by the American Society of Clinical Oncology and the European Group on Tumor Markers 7 as a prognostic biomarker for recurrent CRC following curative resection. Aims: To study the distribution of carcinoembryonic antigen in colorectal carcinoma, the role of carcinoembryonic antigen in the prognosis of colorectal carcinoma, to the variability of carcinoembryonic antigen values in recurrence of colorectal carcinoma and the sensitivity and specicity of carcinoembryonic antigen estimation in colorectal cancer Material And Method: This Institution based observational and prospective study was all the adult patients(&gt;18yrs) who have been diagnosed clinically with colorectal carcinoma and with recurrent disease in the Department of General surgery of R.G.Kar medical college and hospital from January 2019 to August 2020 were included in this study. Result: 10% of the patients were in Duke stage A, 16% patients were Duke stage B; 40% patients were with Duke stage C and 34%patients were Duke stage D of colorectal carcinoma. Among those 13 patients with Duke A and B staging 30% had increased CEA, whereas among those 37 patients with Duke stage C and D 92% had increased CEA levels. The p-value was found to be 0.00033 that is&lt;0.05,which meant the test was statistically signicant. 80%(n=38) of the patients with colon carcinoma had increased CEAvalues whereas 20%(n=2) of the patients had normal CEAvalues. 77%(n=7) patients in the rectal carcinoma group had increased CEAlevels whereas in 23%(n=3) patients it was normal. Conclusion:This study shows an increased amount of CEAlevels in the preoperative period then the patient's chances of recurrence and metastasis increases, hence having a poor prognosis. Regular surveillance through serial monitoring of the CEAlevels in the postoperative period is essential to be vigilant of recurrences and metastasis, which may increase chances of mortality in the patient.

TEAD4 nuclear localization and regulation by miR-4269 and miR-1343-3p in colorectal carcinoma

Background and aimsTEAD4 transcription factor belonging to TEAD-family, is a key downstream element of the Hippo Signalling pathway and is very important for YAPinduced tumor progression. YAP-TEAD interaction is required to promote tumor progression and metastasis in various cancers. This study aims to investigate the role of TEAD4 in CRC progression and to compare the TEAD4 expression with different clinicopathological parameters of the study population. We also aim to explore the expression pattern of miR-4269 and miR-1343–3p and their functional role in TEAD4 mediated CRC progression. Furthermore, we intend to evaluate the prognostic significance of TEAD4, miR-4269, and miR-1343–3p in colorectal carcinoma. MethodsReal-time PCR, Immunohistochemical Staining, and Western Blotting were performed on 71 human CRC tissue specimens and their adjacent normal tissues to evaluate the TEAD4 expression and the results were statistically analyzed against the clinicopathological variables of patient data and also with survival data using STATA software. miRNA expression was analyzed by quantitative real-time PCR. ResultsTEAD4 expression levels in tumor specimens were significantly higher than their paired normal specimens. The higher protein expression levels showed a significant association with TNM stage, Duke Stage, tumor grade, invasion depth, node status, necrosis of tumor tissue, lymphovascular and perineural invasion. As per the cox-regression model and classification tree analysis, TNM stage and perineural invasion were important predictors for TEAD4 expression and prognosis of CRC patients. Survival analysis indicated that TEAD4 overexpression was associated with poorer overall and disease-free survival. miR-4269 and miR-1343–3p were downregulated in CRC tumors and showed a negative correlation with TEAD4. The nuclear overexpressed TEAD4 and downregulated miR-4269 and miR-1343–3p evaluated for the first time in CRC, are believed to serve as important prognostic markers in CRC. ConclusionExpression of TEAD4 was increased in CRC and was negatively regulated by miR-4269 and miR-1343–3p. The overexpression of TEAD4 is linked with poor overall and disease-free survival of CRC patients. These findings support prior observations and thus TEAD4 may be a possible prognostic marker in CRC.

Prevalence and Management of Cancer of the Rectal Stump after Total Colectomy and Rectal Sparing in Patients with Familial Polyposis: Results from a Registry-Based Study.

Simple SummaryProphylactic total colectomy with ileo-rectal anastomosis (IRA) is the surgical approach that guarantees a better quality of life compared to proctocolectomy and ileo-anal anastomosis (IPAA) in familial adenomatous polyposis (FAP) patients. However, previous studies have warned about the high risk of cancer of the rectal stump, especially considering the young age of these patients. This is a retrospective study whose aim was to assess both clinical and surgical features of patients who developed cancer of the rectal stump. The data reported show that IRA is a safe approach from an oncological perspective. Since early tumors of the rectal stump may be easily detected via endoscopic or minimally invasive approaches, strict surveillance is necessary.Background: The balance between quality of life and colorectal cancer risk in familial adenomatous polyposis (FAP) patients is of primary importance. A cut-off of less than 30 polyps under 1 cm of diameter in the rectum has been used as an indication for performing ileo-rectal anastomosis (IRA) in terms of lower rectal cancer risk. This study aimed to assess clinical and surgical features of FAP patients who developed cancer of the rectal stump. Methods: This retrospective study included all FAP patients who underwent total colectomy/IRA from 1977 to 2021 and developed subsequent rectal cancer. Patients’ features were reported using descriptive statistics by considering the overall case series and within pre-specified classes of age (<20, 20–30, and >30 years) at first surgery. Results: Among the 715 FAP patients, 47 (6.57%, 95% confidence interval: 4.87; 8.65) developed cancer in the rectal stump during follow-up. In total, 57.45% of the population were male and 38.30% were proband. The median interval between surgery and the occurrence of rectal cancer was 13 years. This interval was wider in the youngest group (p-value: 0.012) than the oldest ones. Twelve patients (25.53%) received an endoscopic or minimally invasive resection. Amongst them, 61.70% were Dukes stage A cancers. Conclusions: There is a definite risk of rectal cancer after total colectomy/IRA; however, the time interval from the index procedure to cancer developing is long. Minimally invasive and endoscopic treatments should be the procedures of choice in patients with early stage cancers.

Expression and Clinical Significance of Micro Ribonucleic Acid-132 and Sex-Determining Region Y-Box 4 in Colon Cancer

This study aimed to investigate the expression and clinical significance of micro ribonucleic acid-132 and sex-determining region Y-box 4 gene in colon cancer. Luciferase reporter system was used to validate the target gene directly regulated by micro ribonucleic acid-132. Transwell invasion assay was used to test the effect of micro ribonucleic acid-132 on invasiveness of SW620 cells. Micro ribonucleic acid-132 and sex-determining region Y-box 4 expression levels in cancerous and peri-cancerous tissues were measured using in situ hybridization and EleVisionTM immunohistochemistry staining. Micro ribonucleic acid-132 downregulated sex-determining region Y-box 4 protein expression and inhibited the invasiveness of colon cancer cells. In the 72 cancerous tissue samples of colon cancer, micro ribonucleic acid-132 positive rate was 18.1 % (13/72) and sex-determining region Y-box 4 protein positive rate was 73.6 % (53/72), both expressions showed significant differences as compared to peri-cancerous tissues (p<0.01). Micro ribonucleic acid-132 expression was significantly lower in colon cancer tissues with lymph node metastasis than without lymph node metastasis (p<0.01). Sex-determining region Y-box 4 expression was elevated along with Dukes stage increase and higher expression was accompanied by more tumor invasion depth (both p<0.05). Sex-determining region Y-box 4 expression was significantly higher in colon cancer tissues with lymph node metastasis than without lymph node metastasis (p<0.01). Correlation analysis displayed significant negative correlation of micro ribonucleic acid-132 expression with sex-determining region Y-box 4 expression (r=-0.594, p=0.013). Micro ribonucleic acid-132 inhibits invasion and metastasis of colon cancer cell line SW620 via targeting sex-determining region Y-box 4. Low expression of micro ribonucleic acid-132 and high expression of sex-determining region Y-box 4 protein could be important biomarkers for malignant transformation of colonic mucosa and for invasion and metastasis of colon cancer. Measuring expression levels of both molecules is valuable for predicting invasion and metastasis of colon cancer.

Thymosin β4 and β10 are highly expressed at the deep infiltrative margins of colorectal cancer - A mass spectrometry analysis.

Colorectal cancer (CRC) is a complicated tumor, involving several oncogenic signaling pathways, and with a molecular mechanism not fully understood yet. The implication of thymosin β4 (Tβ4) with tumor insurgence and in migration of CRC cells was evidenced in the past with different methodologies, while Tβ10 connection with CRC has been sporadically investigated. This study focused on the implication of both types of thymosin in CRC progression and invasion by analyzing the changes in their levels according to different zones of the tumor, and to Dukes stage and budding index. Tβ4 and Tβ10 were analyzed in deep and superficial tumor samples, and normal mucosa from 18 patients. Concentrations of Tβ4 and Tβ10 have been measured by high-pressure liquid chromatography (HPLC) coupled to electrospray-ion trap mass spectrometry (ESI-IT-MS). MS data were compared by t-test and ANOVA statistical analysis. Identification of thymosin and their proteoforms has been performed by HPLC-high resolution-ESI-IT-MSMS. Both Tβ4 and Tβ10, exhibited intra-tumoral quantitative differences, being upregulated in the deep part of the CRC. They exhibited, moreover, strong association with the Dukes stage and the budding grade, being more concentrated in patients at Dukes stage B and with budding index "2". The results obtained in the present investigation encouraged the hypothesis that the two thymosin are involved in colorectal cancer progression, and in promoting cancer invasion. Thus, they are good candidates to be diagnostic/prognostic biomarkers and therapy targets.