The pathological and immunohistochemical profile of tumor angiogenesis in perforated sigmoid carcinoma–Case report and short literature review

Objective To observe and compare the clinical characteristics of colorectal related multiple primary carcinoma (MPC) and colorectal carcinoma (CRC) for promoting early diagnosis and treatment of colorectal related MPC. Methods Pathological and clinical documents of clearly staged CRC and colorectal related MPC cases from Jul. 1997 to Nov. 2007 were retrospectively analyzed. Results 573 colorectal carcinoma cases were analyzed, including 45 MPC (7.85 %). Parenteral multiple carcinoma originated most frequently from stomach, and then breast, ovary, lung, small intestine and other sites. Among all multiple primary colorectal carcinomas (MPCC), ascending colon carcinoma was most frequent (34.0 %).While among CRC cases, rectal cancer cases was most frequent(36.5 %). Comparing CRC and MPCC, there were no significant difference in terms of tumor family history. Median morbidity age was 57 years and 63 years respectively. Cases with previous colonic polyps accounted for 20.0 % of all MPC cases, while only 0.9 % of all CRC cases. The mOS of CRC and MPC was 93.7 month and 64.8 month respectively. Most frequent pathological type of CRC and MPC were both well-moderately differentiated adenocarcinoma, but more mucinous adenocarcinoma cases were observed in MPC. Conclusion Colorectal related MPC are relatively common among colorectal carcinoma patients. More patients with MPC especially MPCC has colonic polyp. mOS of MPC is shorter than that of CRC, indicating the poor prognosis of MPC compared with CRC.MPCC has multiple colonic polyps, shorter interval of secondary carcinomas, and shorter mOS, worse prognosis than MPC with parenteral tumor. Key words: Neoplasms, multiple primary; Colorectal neoplasms; Clinical feature

Evaluation of the relationship between clinicopathological and immunohistochemical risk factors for liver metastasis, including CD10 expression, is meaningful in colorectal carcinoma (CRC). The purpose of the present study was to clarify what kind of risk factors are significant and independent factors for the development of liver metastasis in CRC. Sixty cases of advanced CRC with synchronous liver metastasis and sixty cases of advanced CRC without liver metastasis at least 5 years after resection of the primary CRC were randomly selected. We analysed the clinicopathological factors and the expression of four biological factors, CD44, transforming growth factor alpha (TGF-alpha), vascular endothelial growth factor (VEGF) and CD10, by immunohistochemistry. Univariate analysis revealed that the incidence of vascular invasion, the incidence of lymph node metastasis and the expression of CD44, TGF-alpha, VEGF and CD10 were all significantly higher in the cases of CRC with liver metastasis than in cases of non-metastatic CRC. Logistic regression analysis showed that lymph node metastasis, the expression of CD10 and the expression of VEGF were significant factors and independent of the other variables. If all three markers are positive, the probability of liver metastasis becomes 92.7%. In this study, lymph node metastasis, CD10 and VEGF were all found to be significant risk factors for the development of liver metastasis in the cases of CRC. These risk factors according to multivariate analysis are candidate markers for predicting the development of liver metastasis.

To determine whether DNA analysis can be performed using the supernatants of body fluids after centrifugation at 2,000 rpm for 10 minutes, peritoneal or pleural effusions or bile were examined for K-ras mutations in 34 cases of pancreatic, colorectal, gastric, esophageal, or hepatocellular carcinoma and 15 noncancer cases. The polymerase chain reaction products for K-ras gene codons 2 to 97 of exons 1 and 2 were generated with 41 (93%) of 44 body cavity fluid and 5 (100%) of 5 bile samples. By the single strand conformation polymorphism method, point mutations were detected in the ascites supernatants of 8 (89%) of 9 cases of pancreatic carcinoma. In the remaining case, no point mutation was demonstrated because few malignant cells were present in the ascites fluid. Furthermore, K-ras point mutations were observed in the ascites supernatants of 2 cases of colorectal carcinoma and 1 case of gastric carcinoma. The DNA analysis of the supernatant of ascites fluid showed a K-ras point mutation in 3 cases of false-negative cytologic diagnosis (2 cases of pancreatic carcinoma and 1 case of colorectal carcinoma). Direct sequencing confirmed identical point mutations in the supernatants, whole cell pellets, malignant cells from the cytologic smears of ascites fluid, and cancer tissues. This novel method allows simultaneous testing for genetic abnormalities in supernatants of body fluid, after removing cells for cytologic diagnosis.

To evaluate epidermal growth factor receptor and Ki-67 immunohistochemical expression in colorectal adenoma and carcinoma cases, and to relate their expression with the available clinicopathological data. The retrospective study was conducted at the Faculty of Medicine, Kafrelsheikh University, Egypt, from September 2019 to October 2020, and comprised formalin-fixed and paraffin-embedded specimens related to cases of colorectal adenoma and those of colorectal carcinoma, who had no previous radiation or chemotherapeutic treatment. Immunohistochemical staining of all thecases was done using anti-epidermal growth factor receptor and anti-Ki-67 antibodies. Data was analysed using SPSS 20. Of the 70 cases, 20(28.5%) were of colorectal adenoma; mean age 52.95±13.47 years, and male-to-female ratio 1:1. The remaining 50(71.5%) cases had colorectal carcinoma; mean age 51.08±13.49 years, and male-to-female ratio 1.17:1. Epidermal growth factor receptor and Ki-67 overexpression related significantly to villous histopathological type and high-grade dysplasia in colorectal adenoma cases(p<0.05). In colorectal carcinoma cases, epidermal growth factor receptor overexpression related significantly to tumour grade (p<0.05). Ki-67 overexpression related significantly to increased pathological stage (p<0.05). Overexpression of epidermal growth factor receptor and Ki-67 was found to be an ominous sign of colorectal adenoma aggressiveness, and the risk of progression to colorectal carcinoma.

A principal molecular mechanism by which cells respond to hypoxia is by activation of the transcription factor hypoxia-inducible factor 1alpha (HIF-1alpha). Several studies describe a binding of p53 to HIF-1alpha in a protein complex, leading to attenuated function, half-life, and abundance of HIF-1alpha. However, these reports almost exclusively utilized transformed cell lines, and many employed transfection of p53 or HIF-1alpha plasmid constructs and/or p53 and HIF-1alpha reporter constructs as surrogates for endogenous protein activity and target expression, respectively. Thus, it remains an open and important question as to whether p53 inhibits HIF-1alpha-mediated transactivation of endogenous HIF-1alpha targets in nontransformed cells. After determining in primary astrocyte cultures the HIF-1alpha targets that were most dependent on HIF-1alpha function, we examined the effect of the loss of p53 function either alone or in combination with MDM2 on expression of these targets. Although p53 null astrocyte cultures resulted in markedly increased HIF-1alpha-dependent target expression compared with controls, this altered expression was determined to be the result of increased cell density of p53 null cultures and the accompanying acidosis, not loss of p53 protein. Although activation of p53 by DNA damage induced p53 target expression in astrocytes, it did not alter hypoxia-induced HIF-1alpha target expression. Finally, a combined loss of MDM2 and p53 did not alter HIF-1alpha target expression compared with loss of p53 alone. These data strongly suggest that p53 and MDM2 do not influence the hypoxia-induced transactivation of HIF-1alpha targets, regardless of p53 activation, in primary astrocytes.

Objectives The aim of this work was to study the immunohistochemical expression of CD44 in colorectal carcinoma (CRC) cases. Background Cancer stem cells (CSCs) have a role in cancer initiation, progression, and metastasis of CRC through their ability of self-renewal and unlimited proliferation, and they seem to be responsible for local relapse and metastasis. Several markers for CSCs have been investigated in CRC, and CD44 was the most likely marker for colorectal CSCs. Materials and methods This retrospective study included 71 colorectal specimens (49 CRC, 13 adenoma, and nine normal cases). The studied cases were collected from the Pathology Department, Faculty of Medicine, Menoufia University, during the period from 2011 to 2015. All cases were stained with CD44 antibody. Survival data were available for 31 of 49 studied CRC cases. Results All cases of normal colonic epithelium and normal colon adjacent to CRC showed no membranous CD44 immunoreactivity; 84.6% of the studied adenoma cases showed no expression of CD44, whereas 57.1% of CRC cases showed positive expression. There was a statistically significant association between positive epithelial expression of CD44 and left-sided tumors (P = 0.01). There was a statistically significant difference between CRC and colorectal adenoma (P = 0.007) and between CRC and normal cases (P = 0.000) as regards CD44 expression. There was a highly statistically significant association between stromal expression of CD44 and absence of lymph node invasion (P = 0.002) as well as early Dukes' stage (P = 0.01). Conclusion CD44 may be involved in the pathogenesis of CRC. Stromal expression only of CD44 is associated with better prognostic factors.

p53 aerobics: The major tumor suppressor fuels your workout

‘Orphan arteriole’ sign – a helpful clue in detecting extramural venous invasion in colorectal carcinomas

The endoplasmic reticulum (ER) is the central intracellular organelle responsible for proper folding, localization, and post-translational modifications of proteins in eukaryotic cells. During conditions of cellular stress, incorrectly folded proteins can accumulate in the ER and trigger the unfolded protein response (UPR), which involves the activation of signal transduction pathways designed to restore correct protein folding and recovery from ER stress [1]. In rapidly dividing tumor cells, UPR can become constitutively activated, presumably because higher levels of proliferation require increased protein synthesis [1, 2]. In addition, the tumor micro-environment is often host to additional adverse conditions including nutrient and oxygen deprivation, leading to further perturbations of ER function. Therefore, it may come as no surprise that UPR has been shown to play a crucial cytoprotective role in tumor cells [2]. We have recently unveiled an unexpected role for the tumor suppressor p53 in regulating ER function via the IRE1α/XBP1 pathway [3]. IRE1α is an ER stress sensor that activates the UPR to restore ER function and maintain cellular homeostasis. It was observed that wild-type p53 suppresses IRE1α expression and activation, while the loss of active p53 potentiates the IRE1α/XBP1 pathway, enhancing ER function [3]. This finding suggests a survival benefit for cancer cells that can inactivate p53. Because cancer cells with high protein load on the ER would need enhanced ER function to cope with constant ER stress, losing p53 function would assist tumors to grow in adverse conditions by increasing IRE1α/XBP1 pathway activation. In addition, we found that a p53 target, CDIP1, acts as a proapototic signal mediator in conditions of ER stress [4]. The CDIP1/BAP31 complex was shown to transduce ER stress-mediated apoptotic signaling to the mitochondria and activate the intrinsic apoptosis pathway, suggesting that loss of p53 function may contribute to the survival of cancer cells by enhancing ER function as well as hindering ER stress-mediated apoptosis signaling. The loss of p53 function in relation to ER stress is likely to have important consequences for the sustenance of cancer cell proliferation. Previous studies have demonstrated that the activation of p53 suppresses mTOR activity and consequently inhibits protein translation [5, 6]. The elimination of wild-type p53 function may therefore provide an opportunity for cancer cells to further promote unregulated proliferation, via elevated protein synthesis and a subsequent increase in ER function. We have revealed that wild-type p53 directly interacts with synoviolin (SYVN1) and can thereby stimulate SYVN1-dependent degradation of IRE1α. The loss of p53 disrupts SYVN1-mediated IRE1α downregulation, producing an ER stress-resistant phenotype by increased IRE1α/XBP1 pathway activity [3]. However, although the loss of p53 function facilitates enhanced ER function to manage high ER stress, this also generates an overreliance on IRE1α in cancer cells with p53 mutations. Indeed, we have found that pharmacological inhibition of IRE1α strongly suppresses protein secretion and potently induces cell death in such p53-deficient cancer cells [3]. Importantly, the IRE1α inhibitor exhibited significant inhibitory effects against p53-deficient human tumors in vivo compared to those with wild-type p53. These results illuminate an important proof-of-concept for targeting tumors with mutant p53 via inhibition of the IRE1α/XBP1 pathway. Malignant transformation occurs through a combination of oncogenic pathway activation and the loss of tumor suppression. Understanding the consequences and identifying vulnerabilities caused by such changes can provide promising opportunities to selectively target cancer cells. Mutation in the p53 gene is the most frequent genetic alteration found in human cancer and inactivation of p53 is known as one of the strongest driving factors in cancer development and chemoresistance [7]. The new finding that loss of p53 activates the IRE1α/XBP1 pathway leading to enhanced ER function broadens our understanding of how cancer cells with mutant p53 survive, and also highlights a possibility for the development of novel therapeutic strategies.

Loss of the Ras Regulator RASAL1: Another Route to Ras Activation in Colorectal Cancer

Specific Detection of Cytokeratin 20-Positive Cells in Blood of Colorectal and Breast Cancer Patients by a High Sensitivity Real-Time Reverse Transcriptase-Polymerase Chain Reaction Method

Background: Serrated adenocarcinoma (SA), a subtype of colorectal carcinoma, and the KRAS mutation, a strong marker for the patient’s response to anti-epidermal growth factor receptor therapy, have a clinical importance because of its progressive nature and tendency for chemoresistance. The purposes of this study were to (1) determine the prevalence of SA, (2) evaluate the histomorphological characteristics of SA and classical adenocarcinoma based on its prognostic factors, (3) determine the prevalence of the KRAS mutation in SA cases, and (4) identify the main characteristics of SA cases and classical adenocarcinoma with a KRAS mutation.Methods: This study was conducted by reviewing hematoxylin-eosin-stained slides of colorectal carcinoma (CRC) cases from January 2013 to July 2015 at the Department of Anatomical Pathology Cipto Mangunkusumo General Hospital. The final diagnosis of SA was based on the Tuppurainen et al criteria and the KRAS mutation was evaluated using real-time polymerase chain reaction.Results: Among the 117 adenocarcinoma cases, there were 41 unequivocal SA, 11 equivocal SA, and 65 classical adenocarcinoma. The prevalence rates of unequivocal and equivocal SA among all CRC cases were 7.7% and 2.1%, respectively. There were 11 (28.2%) cases of wild-type KRAS and 28 (71.7%) cases of mutated KRAS among all unequivocal SA cases. Tumor budding (TB) was the predominant prognostic factor.Conclusion: The prevalence of SA among all CRC cases was 7.7%. The KRAS mutation was found in almost three-quarters of all SA cases.

Since early detection increases survival rates, colorectal carcinoma (CRC) is a major concern for researchers. CD10 is a cell membrane-bound metalloproteinase involved in carcinogenesis. Studies have associated it with the progression of CRC to advanced stages, metastasis, and venous invasions. We aimed to evaluate the immunohistochemical expression of CD10 in the tumor and stromal cells of colorectal adenoma and CRC and its correlation with the pathological prognostic factors. This cross-sectional study was conducted on radical resection specimens of CRC and polypectomy specimens of the colorectal adenomas received for routine histopathological evaluation in the Department of Pathology, Ramaiah Medical College and Hospitals, Bengaluru, from March 2021 to October 2022. Tumor morphology was examined by light microscopy, and CD10 expression was evaluated by immunohistochemistry. Descriptive statistics in terms of percentage and Chi-square test/ Fisher exact tests were used for the statistical analysis. The study includes 46 cases of adenomas and CRCs each. Stromal CD10 expression was significantly higher in the carcinomas (63.4%) than in the adenomas (41.3%). Proliferative CRCs showed a significantly higher tumoral CD10 expression. The increase in the stromal CD10 expression in CRCs with increasing grades was found to be statistically significant. No significant association was seen between CD10 expression and other factors. The results indicate a potential role of CD10 in the adenoma-carcinoma sequence. The significant increase in proliferating and high-grade CRCs suggests that CD10 could prove to be a potential biomarker for aggressiveness and also a therapeutic target in CRCs.

The aim of this study was to evaluate cyclooxygenase-2 (COX-2) immunoreactivity in colorectal adenocarcinomas and to find correlations with different pathological features. This study included 35 cases of colorectal carcinoma for which surgical colectomy specimens were collected. Immunohistochemical staining of COX-2 (cyclooxygenase-2) is done by using the Streptavidin-biotin technique. This work reveals that COX-2 is positive in most cases of colorectal carcinoma and negative in normal colon tissue with statistically non significant relations between COX-2 immunostaining and different pathological features. Our data suggest overexpression of COX-2 protein in colorectal carcinoma in contrast to normal mucosa, with a possible role in cell proliferation in carcinogenesis.

Telomere shortening and telomerase activation occur frequently in cases of colorectal carcinoma. In this study, we correlated the clinicopathological parameters with the telomere length in colorectal carcinomas, colonic polyps, and normal colonic tissues. We also investigated whether the telomere length changes reflect the biologic behavior of tumors and different modes of tumor development. Telomere length was determined by terminal restriction fragment Southern blot analysis in 20 invasive colorectal carcinomas and normal mucosa from the same patients. We also examined 20 colonic polyps and associated normal mucosa. Telomere shortening was detected in 16/20 (80%), and telomere elongation in 2/20 (10%) cases of colorectal carcinoma, and no changes in 2 subjects. In the colonic polyp patients, shortening was detected in 4/20 (20%), elongation in 6/20 (30%), and no change in 10/20 (50%). The frequency of telomere shortening was significantly different between colorectal carcinoma and polyp groups. Decreased telomere length was noted in 92.9% (13/14) of Dukes' C and 50% (3/6) of Dukes' B. The difference between these two sub-groups was statistically significant. This study suggests that the telomere length in colorectal carcinomas is decreased upon the development of malignancy. A significant difference in telomere length between polyps and invasive colorectal carcinomas may reflect a different biologic behavior of colorectal carcinomas.