Michał Mik

Matrix Metalloproteinase-2 Cleavage of the β1 Integrin Ectodomain Facilitates Colon Cancer Cell Motility

Background: Cancer cell invasion requires integrins for adhesion/de-adhesion and MMPs for focalized proteolysis. Results: MMP-2 is up-regulated in invasive colorectal tumors and degrades β1 integrins. Conclusion: Shedding of the I-like domain from β1 integrins results in decreased adhesion and enhanced cell motility. Significance: MMP-2 amplifies the motility of cancer cells, not only degrading extracellular matrix but also reducing the β1 integrin expression. Cancer cell invasion is a key element in metastasis that requires integrins for adhesion/de-adhesion, as well as matrix metalloproteinases (MMPs) for focalized proteolysis. Herein we show that MMP-2 is up-regulated in resected colorectal tumors and degrades β1 integrins with the release of fragments containing the β1 I-domain. The β1 cleavage pattern is similar to that produced by digestion of α5β1 and α2β1 with MMP-2. Two such fragments, at 25 and 75 kDa, were identified after immunoprecipitation, with monoclonal antibody BD610468 reacting with the NH2-terminal I-like ectodomain followed by SDS-PAGE and microsequencing using electrospray (ISI-Q-TOF-Micromass) spectrometry. Cleavage of the β1 integrin can be abolished by inhibition of MMP-2 activity; it can be induced by up-regulation of MMP-2 expression, as exemplified by HT29 colon cancer cells transfected with pCMV6-XL5-MMP-2. Co-immunoprecipitation studies of colon cancer cells showed that the β1 integrin subunit is associated with MMP-2. The MMP-2-mediated shedding of the I-like domain from β1 integrins resulted in decreased adhesion of colon cancer cells to collagen and fibronectin, thus abolishing their receptivity. Furthermore, such cells showed enhanced motility as evaluated by a “wound healing-like” assay and time-lapse microscopy, indicating their increased invasiveness. Altogether, our data demonstrate that MMP-2 amplifies the motility of colon cancer cells, not only by digesting the extracellular matrix components in the vicinity of cancer cells but also by inactivating their major β1 integrin receptors.

A/G Polymorphism of the MMP-7 Gene Promoter Region in Colorectal Cancer

In gastrointestinal malignancies increased expression of matrilysin - MMP-7 - is often observed. Its high level positively correlates with clinical stage of malignancy and is a negative prognostic factor. This suggests a possible relationship between functional polymorphisms of the MMP-7 gene and susceptibility to development of colorectal cancer and an aggressive course of the disease.The aim of the study was to assess the effects of A/G functional polymorphism at -181 site of the MMP-7 gene promoter region on development and progression of colorectal cancer.Material and methods. In total, 184 patients treated surgically for colorectal cancer at the Department of General and Colorectal Surgery of the Medical University in Łódź in the years 2006-2009 and a control group of 205 cancer-free individuals with a negative family history for malignancy have been investigated. Polymorphic variants of the MMP-7 gene promoter region have been analysed using the RFLP-PCR method.Results. A statistically significant difference in distribution of genotypes has been found between the investigated group and the control group, and the OR analysis confirmed a relationship between the A/G [1.67 (1.03-2.72); p= 0.038] and G/G [2.12 (1.34-3.38); p = 0.018] genotypes and an increased risk of colorectal cancer. The risk of lymph node involvement was more than twice higher for the G/G genotype (OR = 2.83 (1.18-6.79); P = 0.017). In addition, the analysis of genotype distribution in patients divided into groups according to the T parameter of the TNM classification revealed a relationship between the G/G genotype and advanced tumour infiltration. No relationship between the investigated A/G polymorphism and the presence of distant metastases has been found.Conclusions. Obtained results indicate a possible relationship between -181 A/G polymorphism of the MMP-7 gene and malignant transformation of colorectal epithelial cells and progression of colorectal cancer. This suggests applicability of this polymorphism as a predisposing factor for the disease and a prognostic factor, which in the future may be useful in the management algorithm for colorectal cancer.

Resection of the primary tumour or other palliative procedures in incurable stage IV colorectal cancer patients

Objective  The aims of the study were to analyse the early and late results of surgical treatment in patients with stage IV colorectal cancer (CRC) and to evaluate the effect of primary tumour resection and other clinical factors on survival.

Incidence of colorectal cancer in Poland in 1999-2008.

Introduction Malignant neoplasm of the colon is one of the most common gastrointestinal cancers and takes the second place in terms of incidence in the world. In Asian countries compared with Western countries the incidence is a bit lower. In recent years in Poland there has been a disturbing increase in the incidence of this cancer, particularly in the voivodships Mazowieckie, Slaskie, and Wielkopolskie. Material and methods Statistical data from the National Cancer Registry on the incidence of colorectal cancer in Poland in 1999-2008, including the provinces which are grouped into provinces of Eastern, Western and Central Poland. We analysed data on both men and women, with the division of colon cancer, rectal folds esico and rectum. The analysis took into account the recognized incidence in absolute numbers and age-standardized incidence rates. Results The incidence of colon cancer in 1999 was 3438 cases among men and 3476 women, while in 2008 this number increased in both men and women and for men was 4763, and 4340 for women. In all Polish provinces, in 1999, 2165 men and 1719 women, and in 2008, 3188 men and 2150 women suffered from rectal cancer. Conclusions In the years 1999-2008 there was an increase in incidence of cancer of the colon. In Poland, there are territorial differences in the incidence of colorectal cancer described by the standardized incidence ratio. The incidence in Western and Central Poland is generally higher than for Eastern Poland. Probably, these differences have multiple bases.

Alterations of Chk1 and Chk2 expression in colon cancer

Background and aimsCheckpoint kinases 1 and 2 (Chk1 and Chk2) are emerging as key mediators in diverse cellular responses to genotoxic stress, guarding the integrity of the genome. Recent studies suggest the fundamental role of Chk1 and Chk2 in the network of genome surveillance pathways which coordinate cell cycle progression with DNA repair and cell survival or death. Defects in these two serine/threonine kinases are suggested contributors to the development of both hereditary and sporadic human cancer. Little is known about physiologic activities of Chk1 and Chk2 in the colorectal cancer or their role in tumorigenesis.Patient/methodsExpression of Chk1 and Chk2 and their phosphorylated, i.e., active forms (pChk1, pChk2) was examined by Western blot and ELISA analysis in colorectal carcinomas and normal colonic mucosa.Results/findingsExpression of Chk2 and pChk2 was noted to be decreased in around 50% of studied cancer cases. Quantitative studies of phosphorylated Chk2 revealed significant decrease of pChk2 in early stages of colorectal carcinomas. Furthermore, tumor invasion to local lymph nodes was found to correlate with the increase of pChk2 pool in the studied cases.Interpretation/conclusionReduced expression of Chk2 and activated Chk2 may be an important inactivating mechanism, contributing to the development of colorectal neoplasm. However, during progression of neoplasia, activated Chk2 may contribute to the invasiveness of tumor.

Right- and left-sided colon cancer – clinical and pathological differences of the disease entity in one organ

Introduction Some researchers suggest that cancers located in the right vs. the left side of the colon are different and they can be regarded as distinct disease entities. The aim of this study was to analyze differences in clinical, epidemiological and pathological features of patients with right-sided (RCC) and left-sided (LCC) colon cancer. Material and methods One thousand two hundred and twenty-four patients were operated on due to colorectal cancer. A group of 477 patients (254 women, mean age 65.5 ±11 for the whole group) with colon cancer was included (212 RCC vs. 265 LCC). Results Right colon cancer patients were older (67.8 ±11.3 vs. 63.2 ±11.2; p = 0.0087). Left colon cancer patients underwent surgery for urgent indications more often (17.0% vs. 8.5%; p = 0006). Tumor diameter was greater in the RCC group (55 ±60 mm vs. 38 ±21 mm; p = 0.0003). Total number of removed lymph nodes was higher in the RCC group (11.7 ±6 vs. 8.3 ±5; p = 0.0001). Lymph node ratio was higher in the LCC group (0.45 ±0.28 vs. 0.30 ±0.25; p = 0.0063). We found a strong positive correlation between tumor diameter and the number of removed lymph nodes in the LCC group (r = 0.531). Conclusions These differences may result from the fact that RCC patients are diagnosed at an older age. The smaller number of removed lymph nodes in LCC patients may result in incorrect staging. It is still necessary to find other biological dissimilarities of adenocarcinoma located on different sides of the colon.

Neutrophil to Lymphocyte Ratio and C-Reactive Protein as Two Predictive Tools of Anastomotic Leak in Colorectal Cancer Open Surgery?

Purpose: To assess the prognostic value of postoperative C-reactive protein (CRP) and neutrophil to lymphocyte ratio (NLR) in the development of anastomotic leak (AL) in patients after surgery for colorectal cancer (CRC). Methods: Patients operated on for CRC between 2010 and 2014 were enrolled into the study. The sensitivity, specificity, positive predictive value (PPV) and negative predictive values (NPVs) were calculated for the CRP and NLR measured on the 4th postoperative day (POD). Results: Among 724 patients, AL was diagnosed in 33 (4.6%). The accuracy of CRP in the detection of AL using area under curve was 0.83 with the optimal cut-off value of 180 mg/L, sensitivity 75%, specificity 91%, PPV 52% and NPV 87%. Also, NLR on POD4 was higher in the AL group: 9.03 ± 4.13 vs. 4.45 ± 2.25; p = 0.0012; sensitivity 69%, specificity 78%, PPV 49%, NPV 88% at cut-off point of 6.5. Moreover, CRP and NLR on POD4 were significantly higher in patients who died in the postoperative period: 239 ± 24 mg/L vs. 199 ± 41 mg/L; p = 0.034 and 10.71 ± 2.08 vs. 8.65 ± 4.67; p = 0.029, respectively). Conclusions: CRP and NLR on POD4 possess the ability to predict the development of AL and postoperative mortality after CRC operation. Based on our results, high NPV might be indicative of patients with low risk of AL in their postoperative period.

Risk factors of 30-day mortality following surgery for colorectal cancer.

UNLABELLED The 30-day mortality is one of the factors reflecting the quality of treatment. All these efforts focused on decreasing 30-day mortality will directly improve quality of care. The aim of the study was to identify risk factors of 30-day postoperative mortality in a cohort of patients operated on for colorectal cancer in one tertiary colorectal centre. MATERIAL AND METHODS Patients operated on due to colorectal cancer (CRC) between 2008 and 2014 were included in the study. 30-day mortality was assessed as an endpoint of the retrospective study. All records were collected from prospective database. RESULTS 1744 patients were operated on due to CRC. The 30-day mortality was noted in 65 patients (3.5%). In multivariable analyses we revealed that spread disease and poor general condition at admission were risk factors of 30-day mortality: OR 2.35; 2.01-2.57 95%CI, p=0.03 and OR 2.18; 1.95-2.41 95% CI; p=0.01, respectively. Emergency surgery significantly increased the risk of 30-day mortality: OR 2.64; 2.45-2.87 95%CI; p=0.009. Low serum albumin concentration level and diabetes mellitus were additional risk factors for 30-day mortality, OR 1.65; 1.52-1.78 95%CI; p=0.01 and OR 1.67; 1.41-1.82 95%CI; p=0.03, respectively. Mortality was significantly higher after resection procedures than after only palliative operations: 4.21% vs 1.57%; p=0.002. CONCLUSIONS Emergent patients, patients with advanced disease and in poor general state have to be assessed by multidisciplinary team to prepare them to operation. Additionally to reduce the risk of 30-day mortality decision of extend of surgery should be made by experienced surgeons.

Does C‐reactive protein monitoring after colorectal resection with anastomosis give any practical benefit for patients with intra‐abdominal septic complications?

This study aimed to assess the influence of the C‐reactive protein (CRP) level on the early outcome after elective colorectal resection.

Surgery in Jehovah's Witnesses - our experience

Introduction Surgeons face a special challenge in treating Jehovahs Witnesses who refuse blood transfusion. Aim To present our surgical experience with this group of patients operated on in our department. Material and methods A retrospective study of 16 unselected Jehovahs Witnesses patients was conducted between October 2004 and February 2012. We analysed gender, age, haemogram before and after surgery, types of surgery, postoperative complications and the need for blood transfusion, and/or other drugs stimulating erythrogenesis. Results Eighty-one percent of patients were women; the average age of all patients was 57.3 years. Mean haemoglobin level, preoperative, postoperative, and on the day of discharge from hospital, was 12.5 g/dl, 9.7 g/dl, and 9.29 g/dl, respectively. Over the same time period, mean red blood cell count was 4.53 mln/µl, 3.58 mln/µl, and 3.37 mln/µl, respectively. Two out of 16 patients agreed to have blood transfusion. Drugs used for erythropoiesis stimulation included rEPO, ferrum, and folic acid. No surgical death was noted. Conclusions We found that abdominal surgery was safe in our small group of Jehovahs Witness patients. However, all Jehovahs Witness patients should be fully informed about the type of procedure and possible consequences of blood transfusion refusal. Two of our patients agreed to blood transfusion in the face of risk of death.