Publications by authors named "Agnieszka Halon"

122 Publications

Morin-5'-Sulfonic Acid Sodium Salt (NaMSA) Attenuates Cyclophosphamide-Induced Histological Changes in Genitourinary Tract in Rats-Short Report.

Pharmaceuticals (Basel) 2021 Feb 26;14(3). Epub 2021 Feb 26.

Department of Pharmacology, Wroclaw Medical University, ul. Mikulicza-Radeckiego 2, 50-345 Wrocław, Poland.

Cyclophosphamide (CPX) exerts toxicity in the urogenital system. The current study was designed to evaluate the effect of morin-5'-sulfonic acid sodium salt (NaMSA) on CPX-induced urogenital toxicity in rats. NaMSA (100 mg/kg/daily) and CPX (15 mg/kg/daily) alone or in combination and 0.9% NaCl (as a control) were given intragastrically for 10 days. Testes and epididymes from male and urinary bladders from male and female rats were evaluated histologically. In testes and epididymes, morphological changes and relative decrease in sperm count were assessed. In urinary bladders edema, hemorrhage and urothelium erosions were described by 0-2 points scoring system. Reproductive score (RS-in total 6 points) and urinary bladder score (BS-in total 6 points) were thereafter calculated. In CPX-receiving group RS (2.7) and BS (3.3) were significantly higher than in the control (0.5 and 0.25 for RS and BS, respectively). Co-administration of NaMSA reversed most of the morphological changes, which was reflected by lower RS and BS score (0.5 and 1.2 for RS and BS, respectively). The preliminary findings suggest that NaMSA may attenuate CPX-induced histological changes in rat urogenital tract.
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http://dx.doi.org/10.3390/ph14030192DOI Listing
February 2021

Endothelin A Receptors Expressed in Glomeruli of Renal Transplant Patients May Be Associated with Antibody-Mediated Rejection.

J Clin Med 2021 Jan 22;10(3). Epub 2021 Jan 22.

Department of Nephrology and Transplantation Medicine, Wroclaw Medical University, 50-367 Wrocław, Poland.

Background: Non-human leukocyte antigen (HLA) anti-endothelin A receptor antibodies are presented as being potentially important, but the expression of the endothelin A receptor in glomeruli (ETA receptor (g+)) has not yet been described. We decided to evaluate the presence and relevance of the ETA receptor in for-cause renal transplant biopsies. The aim of our study was to evaluate the immunoreactivity of the ETA receptor and its significance in patients who underwent a renal transplant biopsy due to the deterioration of transplant function, with detailed characterization of staining in glomeruli.

Methods: The immunohistochemical expression of ETA receptor (ETAR) was analyzed in renal transplant biopsies. Microscopic evaluation was performed on paraffin sections in glomeruli. The analysis was performed using a two-step scale (0: lack of ETAR expression; 1: the presence of ETAR expression-mild to moderate immunoreactivity).

Results: We analyzed 149 patients who underwent renal allograft biopsy after renal transplantation. Positive staining of ETA receptors in glomeruli (ETA receptor (g+)) was noticed in 13/149 (8.7%) patients. Five of these 13 (38.5%) patients with ETA receptor (g+) developed antibody-mediated rejection (AMR), while 13 of the remaining 136 (9.5%) ETA receptor (g-) patients developed AMR ( = 0.0022). Graft loss was noticed in all but one ETA receptor (g+) patient with AMR (4/5; 80%), but only in 2/13 (15%) ETA receptor (g-) patients with AMR ( = 0.009) during the first year after biopsy.

Conclusions: The expression of endothelin A receptors in glomeruli seems to be a potentially important feature in the diagnosis of damage during antibody-mediated rejection. It may help to identify patients at a higher risk of allograft rejection and injury.
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http://dx.doi.org/10.3390/jcm10030422DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7865600PMC
January 2021

SNP-SNP Interaction in Genes Encoding PD-1/PD-L1 Axis as a Potential Risk Factor for Clear Cell Renal Cell Carcinoma.

Cancers (Basel) 2020 Nov 26;12(12). Epub 2020 Nov 26.

Laboratory of Genetics and Epigenetics of Human Diseases, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Ul. Weigla 12, 53-114 Wroclaw, Poland.

PD-1/PD-L1 axis plays an important role in maintaining homeostasis and prevention from autoimmunity; however, in the tumor microenvironment, PD-1/PD-L1 interaction is responsible for the evasion of immune surveillance by tumor cells. We therefore hypothesized that single nucleotide polymorphisms (SNPs) in genes encoding PD-1 and PD-L1 molecules are associated with the development and outcome of renal cell carcinoma (RCC). Here we genotyped nine polymorphisms: five of : rs36084323G>A, rs11568821G>A, rs2227981C>T, rs10204525G>A, rs7421861T>C and four of : rs822335C>T, rs4143815G>C, rs4742098A>G, rs10815225G>C in 237 RCC patients (including 208 with clear cell RCC (ccRCC)) and 256 controls, with application of allelic discrimination method with use of TaqMan Assays. Interestingly, we found the SNP-SNP interaction between rs10815225 and rs7421861 polymorphisms associated with ccRCC risk. The rs7421861 TC genotype decreased the risk of ccRCC development compared to TT and CC genotypes in the group of rs10815225 GC + CC individuals (OR = 0.21, CI95% = 0.08; 0.54). While possessing of rs10815225 GC or CC genotype increased susceptibility to ccRCC when compared to rs10815225 GG genotype in individuals with rs7421861 TT or CC genotype (OR = 2.40, CI95% = 1.25; 4.61). In conclusion, genetic variants in and genes, especially taken together as SNP-SNP interactions, can be considered to be ccRCC risk factors.
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http://dx.doi.org/10.3390/cancers12123521DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7760680PMC
November 2020

Colorectal Adenocarcinomas Harboring ALK Fusion Genes: A Clinicopathologic and Molecular Genetic Study of 12 Cases and Review of the Literature.

Am J Surg Pathol 2020 09;44(9):1224-1234

Laboratory of Pathology, National Cancer Institute, Bethesda, MD.

This study determined the frequency and the clinicopathologic and genetic features of colorectal carcinomas driven by oncogenic fusions of the anaplastic lymphoma kinase gene (ALK). Of the 8150 screened tumors, 12 (0.15%) were immunohistochemically ALK-positive with D5F3 antibody. These cancers harbored CAD-ALK (n=1), DIAPH2-ALK (n=2), EML4-ALK (n=2), LOC101929227-ALK (n=1), SLMAP-ALK (n=1), SPTBN1-ALK (n=4), and STRN-ALK (n=1) fusions, as detected by an RNA-based next-generation sequencing assay. ALK fusion carcinomas were diagnosed mostly in older patients with a 9:3 female predominance (median age: 72 y). All tumors, except a rectal one, occurred in the right colon. Most tumors were stage T3 (n=7) or T4 (n=3). Local lymph node and distant metastases were seen at presentation in 9 and 2 patients. These tumors showed moderate (n=6) or poor (n=3) glandular differentiation, solid medullary growth pattern (n=2), and pure mucinous morphology (n=1). DNA mismatch repair-deficient phenotype was identified in 10 cases. Tumor-infiltrating lymphocytes were prominent in 9 carcinomas. In 4 carcinomas, tumor cells showed strong, focal (n=3), or diffuse programmed death-ligand 1 immunoreactivity. CDX2 expression and loss of CK20 and MUC2 expression were frequent. CK7 was expressed in 5 tumors. Four patients died of disease within 3 years, and 7 were alive with follow-up ranging from 1 to 8 years. No mutations in BRAF, RAS, and in genes encoding components of PI3K-AKT/MTOR pathway were identified. However, 1 tumor had a loss-of-function PTEN mutation. Aberration of p53 signaling, TP53 mutations, and/or nuclear accumulation of p53 protein was seen in 9 cases. ALK fusion colorectal carcinomas are a distinct and rare subtype of colorectal cancers displaying some features of mismatch repair-deficient tumors.
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http://dx.doi.org/10.1097/PAS.0000000000001512DOI Listing
September 2020

Histopathological assessment of oral leukoplakia. Osteonectin as possible biomarker for further diagnostics.

Pol J Pathol 2020 ;71(2):138-145

Department of Pathomorphology and Oncological Cytology, Faculty of Dentistry, Wroclaw Medical University, Wroclaw, Poland.

Clinical evaluation of oral leukoplakia (OL), confirmed by the histological evaluation of the suspected area, provides the gold standard for diagnostics of this pathology. The aim of present study was to encrypt the significance of the histopathological results (oral intraepithelial neoplasia - OIN, WHO 2005, Ljubljana classification systems) of OL. The usefulness of osteonectin as a biomarker of changes in the oral cavity epithelium was evaluated. IRS Score to evaluate osteonectin (SPARC - secreted protein acidic and rich in cysteine) production in oral mucous tissues was modified, with the aim of adapting the diagnostic measurements to the OL cell environment. In total, 37 formalin-fixed, paraffin-embedded (FFPE) blocks from patients with clinically diagnosed OL, and 29 FFPE blocks from patients with OSCC were evaluated. The OIN and system from Ljubljana were compared, to adjudicate which was most compatible with WHO 2005 histopathological assessment. Increased production of SPARC was observed, with the progression in severity of pathological changes in the oral mucosa, from simple hyperplasia, through dysplasia, to OSCC. The WHO 2005 and the OIN classification systems can be applied interchangeably.
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http://dx.doi.org/10.5114/pjp.2020.97021DOI Listing
October 2020

Histology of Liver of the Deceased Due to Harmful Use of Alcohol.

Alcohol Alcohol 2020 Aug;55(5):518-523

Department of Forensic Medicine, Wroclaw Medical University, J. Mikulicza-Radeckiego 4 Str., 50-345 Wrocław, Poland.

Aim: To study types and incidence of histological changes in liver of people deceased due to harmful use of alcohol.

Methods: A retrospective review of medico-legal autopsy of 236 adults who died in the years 2015-2016 due to harmful use of alcohol was done. Histopathological liver samples taken during autopsies were evaluated. Blood alcohol content was analyzed. Serological tests for hepatitis B surface antigen and anti-hepatitis C virus (HCV) were performed.

Results: The most common liver pathology (83.1%) was steatosis, mainly mixed type (50%); 66.9% had high-grade steatosis. Liver fibrosis was detected in 39.4% of cases, with fibrosis of higher than or equal to third grade in 14%, hepatitis in 44.5% and steatohepatitis in 19.1%. Toxic hepatocyte injury features (ballooning degeneration, Mallory-Denk bodies) were found in 20.8% cases and degenerative-damage changes in 41.1%. The correlation between the grade of steatosis and fibrosis (P = 0.0005), toxic injury (0.00000101) and degenerative-traumatic changes (P = 0.00000741) was found. The correlation was found between hepatitis and higher than or equal to third grade steatosis (P = 0.037), cholestasis (P = 0.0139), toxic injury features (P = 2.58 × 10-13), degenerative-damage changes (P = 7.9 × 10-12) and presence of anti-HCV (P = 0.00723) and between progression of fibrosis and presence of toxic injury features (2.28 × 10-19), degenerative-damage changes (P = 4.25 × 10-11) and anti-HCV (P = 0.0263).

Conclusions: Spectrum of histopathological liver changes is broad regardless of sex, and various traits are present in various patterns. Comorbidities have strong influence on the picture of changes in the liver. Exact evaluation how often and what histopathological changes will develop in alcohol liver disease is not possible by reason of variability of external factors.
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http://dx.doi.org/10.1093/alcalc/agaa059DOI Listing
August 2020

Clinical Significance of Nucleoli Cytomorphology Assessment in Patients With Uveal Melanoma.

Anticancer Res 2020 Jun;40(6):3505-3512

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University, Wroclaw, Poland.

Aim: To assess the prognostic significance of nucleolar morphological parameters in a large cohort of patients with uveal melanoma.

Patients And Methods: The presence, size and number of nucleoli of cancer cells were assessed in haematoxylin and eosin (HE)-stained slides of 164 formalin-fixed paraffin-embedded primary uveal melanoma tissue specimens. The results were correlated with clinicopathological features and patient survival.

Results: The presence of macronucleoli and multiple nucleoli significantly correlated with the epithelioid type of uveal melanoma, high mitotic rate, and marked pleomorphism. There was a positive correlation between the presence of macronucleoli as well as the number of nucleoli and the largest tumour basal diameter. The increased nucleolus count in tumour cells positively correlated with primary tumour (pT) staging. The presence of both prominent and multiple nucleoli was associated with significantly reduced overall and disease-free survival.

Conclusion: Histological assessment of nucleolar morphology in routine HE staining would be a helpful low-cost method to obtain reliable prognostic information.
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http://dx.doi.org/10.21873/anticanres.14338DOI Listing
June 2020

Angiotensin II Type 1 Receptor Expression in Renal Transplant Biopsies and Anti-AT1R Antibodies in Serum Indicates the Risk of Transplant Loss.

Transplant Proc 2020 Oct 21;52(8):2299-2304. Epub 2020 May 21.

Department of Nephrology and Transplantation Medicine, Wroclaw Medical University, Wrocław, Poland.

The manifestation of anti-angiotensin II type 1 receptor (AT1R) antibodies is considered a risk factor for transplant injury; however, the occurrence of angiotensin II type 1 (AT1)-Receptor expression in renal transplant biopsy may help to predict transplant loss. The aim of our study was to evaluate the expression of AT1-Receptors together with their antibodies and assess the risk of transplant loss in patients who had a renal transplant indication biopsy.

Methods: AT1-Receptor immunoreactivity was analyzed in renal transplant biopsies. Additionally, we analyzed the presence of anti-AT1R antibodies in these patients using the enzyme-linked immunosorbent assay (ELISA) method. A result ≥ 10 was assessed as positive. An immunohistochemical evaluation of AT1-Receptor expression was performed on 4 μm-thick paraffin sections mounted on salinized slides.

Results: We checked 156 samples of biopsies for the immunoreactivity of the AT1-Receptor. Additionally, we analyzed the presence of anti-AT1R antibodies in these patients using the ELISA method. A group of 67 patients had positive AT1-Receptor expression, and 16 patients had positive anti-AT1R antibodies (R+Ab+) results. A group of 89 patients had no expression of AT1-Receptor, among which 51 had also no anti-AT1R (R-Ab-). One-year postbiopsy graft loss in the R+Ab+ patients was 37% (6/16) compared to 10% (7/69) in the R-Ab- patients (P = .006). Two-year and 3-year graft loss was 43% versus 17% and 50% versus 21%, respectively.

Conclusions: The presence of anti-AT1R antibodies in serum together with the expression of AT1-Receptor in transplant biopsies was associated with a significantly higher graft loss. The relevance of AT1-Receptor expression analyzed together with anti-AT1R antibodies should be considered for better transplant immunologic risk assessment.
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http://dx.doi.org/10.1016/j.transproceed.2020.01.126DOI Listing
October 2020

Autoimmune hepatitis and IgG4-related disease: a diagnostic challenge.

Pol Arch Intern Med 2020 08 11;130(7-8):695-696. Epub 2020 May 11.

2nd Department of General and Oncological Surgery, Wroclaw Medical University, Wrocław, Poland

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http://dx.doi.org/10.20452/pamw.15350DOI Listing
August 2020

ROCK1 and ROCK2 Are Down-regulated in Aggressive and Advanced Skin Melanomas - A Clinicopathological Perspective.

Anticancer Res 2020 Apr;40(4):1931-1942

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University, Wroclaw, Poland.

Background: RhoA and its downstream effectors Rho-associated coiled-coil kinases (ROCK) 1 and 2 are central controllers of cytoskeleton dynamics, and therefore influence cell shape, adhesion and migration. Since modulation of these processes holds promise for an effective anticancer strategy, effects of ROCK inhibition have been evaluated in a number of malignancies.

Materials And Methods: Using immunohistochemistry, ROCK1 and ROCK2 expression was semi-quantitatively assessed in 129 patient-derived primary melanomas.

Results: There was a striking predilection for low melanocytic expression of both kinases in thick, ulcerated and mitogenic tumors, as well as in nodular histological type. ROCK1 and -2 expression in tumor-infiltrating lymphocytes (TILs) was preferentially down-regulated in advanced and aggressive tumors. Moreover, diminished ROCK2 reactivity in melanoma cells and TILs was associated with shorter melanoma-specific and recurrence-free survival.

Conclusion: This is the first analysis of ROCK1 and -2 protein expression in clinical melanoma samples and the results indicated the suppression of ROCK signaling in melanocytes of aggressive and late-stage tumors. Functional models that more accurately represent the clinical setting are necessary to dissect the role of ROCK1 and -2 in melanoma. Additionally, our study indicates that ROCK activity in TILs may be involved in the pathogenesis of cancer, and thus merits further investigations.
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http://dx.doi.org/10.21873/anticanres.14148DOI Listing
April 2020

Prevalence and clinical presentation of lymphoproliferative disorder in patients with primary Sjögren's syndrome.

Rheumatol Int 2020 Mar 1;40(3):399-404. Epub 2020 Feb 1.

Department of Rheumatology and Internal Medicine, Medical University of Wroclaw, Borowska 213, 50-556, Wroclaw, Poland.

Lymphomas are one of the serious complications of the primary Sjörgen's Syndrome (pSS). The aim of the study was to evaluate the frequency of lymphoma in pSS. The singe-center retrospective study included 198 Caucasian patients, who met diagnostic criteria for pSS. The type of lymphoproliferative disorder was classified according to the WHO 2016 classification. The mean time of observation, after pSS diagnosis, was 48 weeks. Focus score (FS) ≥ 1 was present in 85% of the patients, and anti-SSA antibodies were detected in 84%. Rheumatoid factor was detected in 130 (65%) patients. Mean disease activity index, according to EULAR Sjörgen's Syndrome disease activity index (ESSDAI), was 8.3 points at the moment of pSS diagnosis. Complement C3 was decreased in 14% of the patients, while 10% showed reduced complement C4. Four patients (2%) were diagnosed with a lymphoma. Most of the patients were diagnosed with mucosa-associated lymphoid tissue lymphoma (MALT), in whom the tumour was located in the parotid gland, and in one patient the stomach was involved. Finally, one patient was diagnosed with a rare B-cell small lymphocytic lymphoma located in the lungs. In this article, we present detailed characteristics of each case. In analysed population the frequency of lymphoma in the course of pSS in patients with pSS is 2%. The variety of lymphoma types in pSS patients imposes individual monitoring in each patient at every check-up visit for disease activity.
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http://dx.doi.org/10.1007/s00296-020-04522-7DOI Listing
March 2020

Filler Migration and Florid Granulomatous Reaction to Hyaluronic Acid Mimicking a Buccal Tumor.

J Craniofac Surg 2020 Jan/Feb;31(1):e78-e79

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University.

Hyaluronic acid is among the most commonly used cosmetic fillers. Although considered biocompatible and safe, it may rarely cause a wide range of complications. The authors report a case of migration of hyaluronic acid concomitant with granulomatous inflammatory response that mimicked a buccal tumor. A 52-year-old female presented with a solid painless mass of the right buccal area. The patient denied any history of trauma and cosmetic procedures of the affected area. Skin and mucosal membrane were intact and the lesion was firm and well fixed in the deep plane. Due to worrisome clinical presentation and the patient's history of breast cancer, the lesion was excised radically. Histopathological examination revealed multiple granulomas surrounding amorphous lakes of hyaluronic acid. During repeated, thorough anamnesis the patient admitted having underwent lip augmentation and nasolabial fold correction with HA two years before, after which the filler must have migrated posteriorly. Physicians need to be aware of various complications associated with cosmetic fillers as they may mimic severe clinical conditions.
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http://dx.doi.org/10.1097/SCS.0000000000005928DOI Listing
March 2020

The effect of YAP expression in tumor cells and tumor stroma on the prognosis of patients with squamous cell carcinoma of the oral cavity floor and oral surface of the tongue.

Oncol Lett 2019 Oct 31;18(4):3561-3570. Epub 2019 Jul 31.

Department of Radiotherapy, Lower Silesian Oncology Centre, 53-413 Wroclaw, Poland.

Classic prognostic factors, such as clinical advancement of the disease and histological grade of the tumor, continue to have a decisive role in the selection of therapeutic strategy in patients with carcinoma of the oral cavity floor and oral surface of the tongue (OCC). YAP1/Yes-associated protein 1 (YAP) and transcriptional co-activator with PDZ-binding motif, WWTR1 (TAZ) proteins, appear to be promising markers that may be used to develop personalized therapies. The aim of the present study was to analyze the associations between the levels of YAP, TAZ and tyrosine-protein phosphatase non-receptor type 14 (PTPN14) and to determine whether the increased expression of YAP and TAZ had an effect on tumor cell proliferation, as determined by minichromosome maintenance 7, DNA replication licensing factor 7 expression. Their prognostic value was also assessed. In total, 127 patients who underwent radical surgery and were subjected to adjuvant radiation therapy due to squamous cell OCC were enrolled in the present study. The results demonstrated an evident effect as YAP expression increased in cancer-associated fibroblasts, which induced unfavorable prognosis in patients. In addition, a positive association between proliferation in cancer cells and YAP expression in stromal cells was observed. A lack of YAP expression in the cytoplasm of tumor cells was a factor for poor prognosis with regard to disease-free survival and disease specific survival. No statistically significant correlations between YAP and TAZ expression and PTPN14 expression were identified, nor was a correlation between cell proliferation and the presence of YAP and TAZ in tumor cells observed. The results indicated that YAP expression levels may support the development of personalized therapies for patients.
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http://dx.doi.org/10.3892/ol.2019.10695DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6757271PMC
October 2019

Colonic Adenocarcinomas Harboring NTRK Fusion Genes: A Clinicopathologic and Molecular Genetic Study of 16 Cases and Review of the Literature.

Am J Surg Pathol 2020 02;44(2):162-173

Laboratory of Pathology, National Cancer Institute, Bethesda, MD.

This study was undertaken to determine the frequency, and the clinicopathologic and genetic features, of colon cancers driven by neurotrophic receptor tyrosine kinase (NTRK) gene fusions. Of the 7008 tumors screened for NTRK expression using a pan-Trk antibody, 16 (0.23%) had Trk immunoreactivity. ArcherDx assay detected TPM3-NTRK1 (n=9), LMNA-NTRK1 (n=3), TPR-NTRK1 (n=2) and EML4-NTRK3 (n=1) fusion transcripts in 15 cases with sufficient RNA quality. Patients were predominantly women (median age: 63 y). The tumors involved the right (n=12) and left colon unequally and were either stage T3 (n=12) or T4. Local lymph node and distant metastases were seen at presentation in 6 and 1 patients, respectively. Lymphovascular invasion was present in all cases. Histologically, tumors showed moderate to poor (n=11) differentiation with a partly or entirely solid pattern (n=5) and mucinous component (n=10), including 1 case with sheets of signet ring cells. DNA mismatch repair-deficient phenotype was seen in 13 cases. Tumor-infiltrating CD4/CD8 lymphocytes were prominent in 9 cases. Programmed death-ligand 1 positive tumor-infiltrating immune cells and focal tumor cell positivity were seen in the majority of cases. CDX2 expression and loss of CK20 and MUC2 expression were frequent. CK7 was expressed in 5 cases. No mutations in BRAF, RAS, and PIK3CA were identified. However, other genes of the PI3K-AKT/MTOR pathway were mutated. In several cases, components of Wnt/β-catenin (APC, AMER1, CTNNB1), p53, and TGFβ (ACVR2A, TGFBR2) pathways were mutated. However, no SMAD4 mutations were found. Two tumors harbored FBXW7 tumor suppressor gene mutations. NTRK fusion tumors constitute a distinct but rare subgroup of colorectal carcinomas.
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http://dx.doi.org/10.1097/PAS.0000000000001377DOI Listing
February 2020

Low RhoA expression is associated with adverse outcome in melanoma patients: a clinicopathological analysis.

Am J Transl Res 2019 15;11(7):4524-4532. Epub 2019 Jul 15.

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University Borowska 213, Wroclaw, Poland.

RhoA GTPase is physiologically involved in the formation of stress fibers, cellular contractility and polarity, maintenance of cell cycle and transcriptional control. During tumorigenesis, it plays roles in cancer cell proliferation, apoptosis, adhesion, invasion and metastasis. While RhoA seems to act as a tumor promotor in most malignancies, data regarding its function in skin melanoma are fragmentary and conflicting. We aimed to clarify the clinical significance of RhoA expression in melanoma by immunohistochemical evaluation of 134 primary tumors and subsequent statistical analysis with clinicopathological profiles of patients. Increased RhoA expression was associated with thinner tumors, higher grade of tumor-infiltrating lymphocytes and lack of disease recurrence. Moreover, we observed a trend towards higher RhoA expression in cases without concurrent metastases. Recurrence-free survival and melanoma-specific survival of patients with high RhoA-expressing tumors were significantly prolonged. Multivariable regression model adjusting for melanoma thickness and status of regional lymph nodes confirmed independent prognostic value of RhoA immunoreactivity. In summary, we found associations between RhoA expression and histopathological phenotype of primary tumors as well as patient survival which suggest a suppressive role of RhoA in skin melanoma.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6684925PMC
July 2019

The immunoreactivity of TGF-b1 in non-alcoholic fatty liver disease.

Folia Histochem Cytobiol 2019 12;57(2):74-83. Epub 2019 Jun 12.

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University, Wroclaw, Poland.

Introduction: Non-alcoholic fatty liver disease (NAFLD) is a common chronic liver disease which becomes a rapidly growing health problem in the Western countries. The development of the disease is most often connected to obesity. NAFLD is also considered as the hepatic manifestation of metabolic syndrome. Transforming growth factor b1 (TGF-b1) plays an important role in the pathogenesis of liver fibrosis, being involved in activation of hepatic stellate cells, stimulation of collagen gene transcription, and suppression of matrix metalloproteinase expression. The objective of the study was to evaluate by immunohistochemistry the expression of TGF-b1 in the liver tissue of NAFLD patients and correlate it with anthropometric, biochemical and routine histological parameters.

Material And Methods: The study group consisted of 48 patients with diagnosed NAFLD. Liver steatosis, NAFLD Activity Score (NAS) and METAVIR score of fibrosis were evaluated in liver biopsies. The immunoreactivity of TGF-b1 was evaluated semi-quantitatively separately in portal, septal, lobular hepatocytic and lobular sinu-soidal liver compartments. The results were analyzed in regard to patients' clinical and biochemical parameters.

Results: Neither steatosis nor NAS correlated with TGF-b1 expression in any liver compartment, whereas METAVIR score of fibrosis was associated with increased immunoreactivity of TGF-b1 in most of the studied liver compartments. TGF-b1 immunoreactivity showed positive correlation with patients' age and its expression in septal compartment disclosed positive correlation with body mass index, and waist and hip circumference. Hyaluronic acid serum level was positively and iron concentration was negatively associated with TGF-b1 ex-pression in the selected consecutive liver compartments.

Conclusions: The immunohistochemical expression of TGF-b1 may be complementary to routine methods of liver fibrosis evaluation.
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http://dx.doi.org/10.5603/FHC.a2019.0008DOI Listing
July 2019

Evaluation of adipose tissue and liver radiodensity in overweight or obese patients with nonalcoholic fatty liver disease.

Pol Arch Intern Med 2019 05 22;129(5):354-356. Epub 2019 Feb 22.

Department of Gastroenterology and Hepatology, Wroclaw Medical University, Wrocław, Poland

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http://dx.doi.org/10.20452/pamw.4457DOI Listing
May 2019

Biological Aggressiveness of Subclinical No-Mass Ductal Carcinoma In Situ (DCIS) Can Be Reflected by the Expression Profiles of Epithelial-Mesenchymal Transition Triggers.

Int J Mol Sci 2018 Dec 7;19(12). Epub 2018 Dec 7.

Breast Unit, Department of Surgical Oncology, Lower Silesia Oncology Center, 53-413 Wroclaw, Poland.

Epithelial-mesenchymal transitions (EMTs) have been recently implicated in the process of cancer progression. The aim of this study was to assess how the preoperative expression patterns of EMT biomarkers correlate with the risk of postoperative invasion in ductal carcinoma in situ (DCIS) found on stereotactic breast biopsies. N-cadherin, Snail1, and secreted protein acidic and rich in cysteine (SPARC) immunoreactivity was observed in 8%, 62%, and 38% of tumors, respectively. Snail1 and SPARC expressions were significantly related to N-cadherin expression and to each other. The postoperative upgrading rate was associated with a positive preoperative expression of all biomarkers. Significance of Snail1 and SPARC persisted in multivariate analysis, but the impact of SPARC on invasion was more significant. When these two EMT triggers were considered together, the risk of invasion did not significantly differ between the subtypes of DCIS with single positive expression (SPARC-/Snail1+ vs. SPARC+/Snail1-). However, it was significantly lower in single-positive DCIS when compared to lesions of a double-positive profile (SPARC+/Snail1+). Moreover, there were no cases in the double-negative DCIS (SPARC-/Snail1-), with foci of infiltrating cancer found postoperatively in residual postbiopsy lesions. In contrast, DCIS with a combined high SPARC and Snail1 expression (intermediate or strong) had an invasive component in 66⁻100% of tumors.
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http://dx.doi.org/10.3390/ijms19123941DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6320898PMC
December 2018

Immunohistochemical and ultrastructural analysis of sporadic inclusion body myositis: a case series.

Rheumatol Int 2019 07 8;39(7):1291-1301. Epub 2018 Dec 8.

Division of Histology and Embryology, Department of Human Morphology and Embryology, Wroclaw Medical University, Chałubińskiego Street 6a, 50-368, Wrocław, Poland.

Sporadic inclusion body myositis (s-IBM) is a progressive, skeletal muscle disease with poor prognosis. However, establishing the final diagnosis is difficult because of the lack of clear biomarkers in the blood serum and very slow development of clinical symptoms. Moreover, most other organs function normally without any disturbance. Here, in patients with this untreatable disease, we have underlined the importance of immunohistochemical and ultrastructural assessment of skeletal muscle in patients diagnosed with s-IBM. The goal of this study was to identify the distribution of specific antigens and to determine morphological features in order to localize pathological protein aggregates, rimmed vacuoles, and loss of myofibrils, which are key elements in the diagnosis of s-IBM. All studied patients were between 48 and 83 years of age and were hospitalized in the Department of Rheumatology and Internal Medicine between 2011 and 2016. Anamneses revealed an accelerated progression of muscle atrophy, weakness of limb muscles, and difficulties with climbing stairs. Based on histopathology and transmission electron microscopy examination, inflammatory infiltrations consisting of mononuclear cells, severe atrophy and focal necrosis of myofibers, splitting of myofilaments, myelinoid bodies and rimmed vacuoles were observed. Primary antibodies directed against CD3, CD8, CD68, cN1A, beta-amyloid, Tau protein and apolipoprotein B made it possible to identify types of cells within infiltrations as well as the protein deposits within myofibers. Using a combination of immunohistochemistry and electron microscopy methods, we were able to establish the correct final diagnosis and to implement a specific treatment to inhibit disease progression.
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http://dx.doi.org/10.1007/s00296-018-4221-zDOI Listing
July 2019

SMAD7 is a novel independent predictor of survival in patients with cutaneous melanoma.

Transl Res 2019 02 27;204:72-81. Epub 2018 Sep 27.

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University, Wroclaw, Poland.

Overexpression of SMAD7-a hallmark inhibitor of transforming growth factor β (TGFβ) signaling-has been documented and related with adverse prognosis in a number of epithelial malignancies, suggesting that it may be responsible for resistance to TGFβ-induced growth arrest of cancer cells. The involvement of SMAD7 in development and progression of malignant melanoma is unclear, and its expression has not been characterized so far at the protein level in clinical melanoma tissue samples. We evaluated SMAD7 expression in 205 skin melanoma primary tumors by immunohistochemistry and correlated the findings with clinicopathological profiles of patients. Melanocytic SMAD7 was evidenced in 204 cases, and the expression pattern was predominantly nuclear. High expression of SMAD7 was positively associated with several features of tumor aggressiveness, for example, presence of ulceration (P < 0.001), higher tumor thickness (P < 0.001), and mitotic rate (P < 0.001), but not presence of regional or distant metastases. Moreover, high SMAD7 expression independently predicted unfavorable outcome: melanoma-specific survival (hazard ratio = 3.16, P < 0.001) and recurrence-free survival (hazard ratio = 2.88, P < 0.001). Taken together, our results underline the importance of TGFβ signaling in cancer and define SMAD7 as a marker of aggressive tumor behavior and adverse clinical outcomes in melanoma patients.
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http://dx.doi.org/10.1016/j.trsl.2018.09.002DOI Listing
February 2019

Diagnostic Difficulties in Primary Pauci-Melanotic Leptomeningeal Melanomatosis.

Eur Neurol 2018 19;80(1-2):68-70. Epub 2018 Sep 19.

Department of Pathomorphology and Oncologic Cytology, Wroclaw Medical University, Wroclaw, Poland.

We present a rare case of primary malignant melanoma of the central nervous system. We underline the difficulties we faced during diagnostic procedures. Finally, postmortem examination revealed the diagnosis of primary pauci--melanotic leptomeningeal melanomatosis.
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http://dx.doi.org/10.1159/000493197DOI Listing
January 2019

Upregulation of FOXP1 is a new independent unfavorable prognosticator and a specific predictor of lymphatic dissemination in cutaneous melanoma patients.

Onco Targets Ther 2018 14;11:1413-1422. Epub 2018 Mar 14.

Lower Silesian Oncology Centre, Wroclaw, Poland.

Background: FOXP1 is a pleiotropic protein that plays important roles in immune responses (B-cell development regulation and differentiation of monocyte), organ development (cardiac valves, lung, and esophagus), and neuronal development. Besides being the primary regulator of normal human tissue development, FOXP1 also plays a role in tumorigenesis. However, the potential value of FOXP1 expression in tumor prognosis remains controversial. FOXP1 expression was assessed in tumor cells (TCs) and stromal cells (SCs) of cutaneous melanomas with the aim of analyzing the associations between FOXP1 expression and clinicopathological characteristics. We believe this article to be the first report analyzing the correlations between FOXP1 expression and clinicopathological, as well as histological, characteristics in melanoma.

Materials And Methods: In total, 96 formalin-fixed, paraffin-embedded primary cutaneous melanoma tissue specimens were subjected to immunohistochemical analysis for FOXP1, and the results were correlated with classical clinicopathological features and patient survival.

Results: FOXP1 overexpression in TCs was strongly associated with the presence of metastases in sentinel lymph nodes (=0.0003, OR=11.66) and positive status of regional lymph nodes (=0.0006, OR=22.15). In 96% (52 of 54) of patients presenting with low FOXP1 expression, no clinical or histopathological features of lymphatic dissemination were observed. However, thinner and nonulcerated tumors were reported to have increased numbers of FOXP1-positive SCs. In addition, a strong association was observed between FOXP1 upregulation in SCs and the absence of regional lymph node metastases. There was a significant correlation between FOXP1 upregulation in TCs and shorter cancer-specific overall survival (log-rank test, =0.0040) and disease-free survival (log-rank test, =0.0021). FOXP1 expression was confirmed in multivariate analysis as a factor that significantly unfavorably impacts prognosis in melanoma patients (HR=3.14, =0.0299, adjusted for age, Breslow thickness, and sex).

Conclusion: The findings from this study indicate that FOXP1 has a major role in melanoma progression, which makes it a candidate for molecular target-based cancer therapy.
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http://dx.doi.org/10.2147/OTT.S151286DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5857151PMC
March 2018

Nuclear pseudoinclusions in melanoma cells: prognostic fact or artifact? The possible role of Golgi phosphoprotein 3 overexpression in nuclear pseudoinclusions generation.

Pathol Int 2018 Feb 29;68(2):117-122. Epub 2018 Jan 29.

Department of Oncology, Wroclaw Medical University, pl. Hirszfelda 12, 53-413 Wroclaw, Poland.

Nuclear pseudoinclusions (NPIs) are classically found in papillary thyroid carcinoma and meningioma. Although NPIs have been described in melanocytic lesions, there is no systematic analysis of potential relationship between NPIs and other clinicopathological characteristics of melanoma. We examined the presence of NPIs in H&E-stained tissue sections form 96 melanomas and analyzed statistical associations with important clinicopathological parameters and tissue immunoreactivity for selected proteins involved in epithelial-mesenchymal transition (SPARC, N-cadherin), cell adhesion and mobility (ALCAM, ADAM-10), regulation of mitosis (PLK1), cell survival (FOXP1) and functioning of Golgi apparatus (GOLPH3, GP73). NPIs were observed in 20% of melanomas and their presence correlated with high mitotic rate and ulceration of the tumor, but not with Breslow thickness, histologic type, or presence of metastases. We observed a significant correlation with shorter cancer-specific survival, but not disease-free survival. Presence of NPIs was related to high expression of GOLPH3 in melanoma cells, whereas their absence was linked to enhanced immunoreactivity of GOLPH3 in tumor-associated macrophages. NPIs are not an uncommon finding in skin melanoma and their diagnostic and prognostic utility could be helpful in the daily routine histopathological practice. The possible explanation of NPI generation is associated with enhanced activity of Golgi apparatus in melanoma cells.
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http://dx.doi.org/10.1111/pin.12629DOI Listing
February 2018

Nucleoli cytomorphology in cutaneous melanoma cells - a new prognostic approach to an old concept.

Diagn Pathol 2017 Dec 29;12(1):88. Epub 2017 Dec 29.

Lower Silesian Oncology Centre, pl. Hirszfelda 12, 53-413, Wroclaw, Poland.

Background: The nucleolus is an organelle that is an ultrastructural element of the cell nucleus observed in H&E staining as a roundish body stained with eosin due to its high protein content. Changes in the nucleoli cytomorphology were one of the first histopathological characteristics of malignant tumors. The aim of this study was to assess the relationship between the cytomorphological characteristics of nucleoli and detailed clinicopathological parameters of melanoma patients. Moreover, we analyzed the correlation between cytomorphological parameters of nucleoli and immunoreactivity of selected proteins responsible for, among others, regulation of epithelial-mesenchymal transition (SPARC, N-cadherin), cell adhesion and motility (ALCAM, ADAM-10), mitotic divisions (PLK1), cellular survival (FOXP1) and the functioning of Golgi apparatus (GOLPH3, GP73).

Methods: Three characteristics of nucleoli - presence, size and number - of cancer cells were assessed in H&E-stained slides of 96 formalin-fixed paraffin-embedded primary cutaneous melanoma tissue specimens. The results were correlated with classical clinicopathological features and patient survival. Immunohistochemical analysis of the above mentioned proteins was described in details in previous studies.

Results: Higher prevalence and size of nucleoli were associated with thicker and mitogenic tumors. All three nucleolar characteristics were related to the presence of ulceration. Moreover, microsatellitosis was strongly correlated with the presence of macronucleoli and polynucleolization (presence of two or more nucleoli). Lack of immunologic response manifested as no TILs in primary tumor was associated with high prevalence of melanoma cells with distinct nucleoli. Interestingly, in nodular melanoma a higher percentage of melanoma cells with prominent nucleoli was observed. In Kaplan-Meier analysis, increased prevalence and amount, but not size of nucleoli, were connected with shorter cancer-specific and disease-free survival.

Conclusion: (1) High representation of cancer cells with distinct nucleoli, greater size and number of nucleoli per cell are characteristics of aggressive phenotype of melanoma; (2) higher prevalence and size of nucleoli are potential measures of cell kinetics that are strictly correlated with high mitotic rate; and (3) high prevalence of cancer cells with distinct nucleoli and presence of melanocytes with multiple nucleoli are features associated with unfavorable prognosis in patients with cutaneous melanoma.
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http://dx.doi.org/10.1186/s13000-017-0675-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5747151PMC
December 2017

Clinical, Histopathological and Cytogenetic Prognosticators in Uveal Melanoma - A Comprehensive Review.

Anticancer Res 2017 12;37(12):6541-6549

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University, Wroclaw, Poland.

Uveal melanoma is the most prevalent primary intraocular cancer in adults. Although it accounts for only 5% of all melanomas, it is responsible for 13% of deaths due to this type of cancer. A wide variety of therapeutic options of primary tumor is available and progress in its management is noticeable. The fact still remains, however, that almost half of patients develop metastases which may be due to practically undetectable cancer spread present as early as at diagnosis of the primary focus. Metastatic disease is uniformly fatal despite systemic therapy. Prediction of metastasis is crucial for prognosis. It also allows targeting of emerging new therapeutic methods to the appropriate group of patients. The Authors reviewed literature concerning epidemiology and etiopathogenesis of uveal melanoma, and its clinical, histopathological and cytogenetic prognosticators.
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http://dx.doi.org/10.21873/anticanres.12110DOI Listing
December 2017

A review on Eph/ephrin, angiogenesis and lymphangiogenesis in gastric, colorectal and pancreatic cancers.

Chin J Cancer Res 2017 Aug;29(4):303-312

2-nd Department of General and Oncological Surgery.

Erythroprotein-producing human hepatocellular carcinoma receptors (Eph receptors) compose a subfamily of transmembrane protein-tyrosine kinases receptors that takes part in numerous physiological and pathological processes. Eph family receptor-interacting proteins (Ephrins) are ligands for those receptors. Eph/ephrin system is responsible for the cytoskeleton activity, cell adhesion, intercellular connection, cellular shape as well as cell motility. It affects neuron development and functioning, bone and glucose homeostasis, immune system and correct function of enterocytes. Moreover Eph/ephrin system is one of the crucial ones in angiogenesis and lymphangiogenesis. With such a wide range of impact it is clear that disturbed function of this system leads to pathology. Eph/ephrin system is involved in carcinogenesis and cancer progression. Although the idea of participation of ephrin in carcinogenesis is obvious, the exact way remains unclear because of complex bi-directional signaling and cross-talks with other pathways. Further studies are necessary to find a new target for treatment.
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http://dx.doi.org/10.21147/j.issn.1000-9604.2017.04.03DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5592818PMC
August 2017

The variety of clinical presentations in IgG4-related disease in Rheumatology.

Rheumatol Int 2018 Feb 30;38(2):303-309. Epub 2017 Aug 30.

Department of Rheumatology and Internal Medicine, Wroclaw Medical University, Borowska 213, 50-556, Wroclaw, Poland.

IgG4-related disease (IgG4-RD) belongs to the group of rare diseases in which the identification of the characteristic histology and immunohistochemistry provides with the gold standard in the diagnosis. The variable organ dysfunction reflects the clinical presentation. The examples of different IgG4-RD presentations in the Rheumatology Unit were discussed in this article. The spectrum of IgG4-RD is wide-ranging and manifested in one or more organs synchronously or metachronously. In the presented article, we described five different cases of IgG4-RD. Four cases were reaffirmed in the histopathological assessment. The clinical and laboratory findings were analyzed and the assigned therapy was discussed. According to our experience, the diagnosis of IgG4-RD requires the careful clinicopathological correlation. The diagnosis relies on the coexistence of various clinical, laboratory, radiological, and histopathological findings, although none of them is pathognomonic itself. The time needed for the diagnosis and variety of clinical forms of IgG4-RD shows that there is need of the cooperation among many specialists for the better and earlier recognition of the disease.
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http://dx.doi.org/10.1007/s00296-017-3807-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5773660PMC
February 2018

Recurrent brown tumor of the vertebral column.

Neurol India 2017 Jul-Aug;65(4):908-909

Department of Neurology, Medical University, Wrocław, Poland.

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http://dx.doi.org/10.4103/neuroindia.NI_1060_16DOI Listing
July 2019

Ki-67 evaluation in breast cancer: The daily diagnostic practice.

Indian J Pathol Microbiol 2017 Apr-Jun;60(2):177-184

Department of Pathomorphology and Oncological Cytology, Wroclaw Medical University, Wroclaw, Poland.

Context: Breast cancer is the most common malignancy in females. It is routinely classified according to the WHO histological typing. However, there is also a molecular classification of breast cancer which is routinely substituted with surrogate subtypes based on expression of estrogen, progesterone, and human epidermal growth factor receptor 2 receptors and proliferation index (PI). PI is defined as the percentage of Ki-67-positive cells among overall cell population. The method commonly applied by pathologists to determine PI is visual scoring of the sample. Strict recommendations for PI assessment do not exist. Thus, the mode of PI evaluation differs significantly between pathologists.

Aims: The aim of our study was to evaluate the daily approach to defining the PI.

Settings And Design: Four practicing nonscholar pathologists were asked to evaluate PI in cases of invasive breast carcinoma.

Subjects And Methods: The study was performed on a group of 98 patients diagnosed with invasive breast carcinoma. Immunohistochemical reaction was performed with monoclonal antibody against human Ki-67 antigen using Ventana BenchMark XT.

Statistical Analysis Used: Results were compared using Pearson's and Spearman's rank correlation coefficients and Fleiss and Cohen's kappa values.

Results: Statistical analysis showed pairwise Pearson's coefficients ranging between 0.77 and 0.84 (P < 0.001) and Spearman's rank correlation coefficients ranging between 0.68 and 0.83 (P < 0.001). The Fleiss kappa value for the 14% cutoff point was 0.58 whereas for the 20% cutoff point was 0.60. The pairwise Cohen's kappa values ranged from 0.45 to 0.69 for the 14% cutoff point and 0.53 to 0.67 for the 20% cutoff point. Friedman's rank ANOVA test showed significant differences among the four pathologists (P < 0.001).

Conclusions: Our study shows a significant difference in results and methods of evaluation of PI between pathologists.
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http://dx.doi.org/10.4103/IJPM.IJPM_732_15DOI Listing
February 2018

Polo-like kinase-1 immunoreactivity is associated with metastases in cutaneous melanoma.

J Cutan Pathol 2017 Oct 19;44(10):819-826. Epub 2017 Jul 19.

Department of Oncology and Division of Surgical Oncology, Wroclaw Medical University, Wroclaw, Poland.

Background: Polo-like kinase-1 (PLK-1) is one of the key regulators of cell cycle progression. Increased expression of PLK-1 was observed in several tumor types.

Methods: We immunohistochemically assessed PLK-1 expression in neoplastic and stromal compartments of 96 cutaneous melanomas, and analyzed associations between PLK-1 expression and clinicopathological characteristics.

Results: PLK-1 expression in cancer cells was not associated with basic clinical (eg, age, gender and tumor location) or histopathological (eg, Breslow thickness, mitotic rate and ulceration) parameters. However, increased PLK-1 was more frequent in tumors with concurrent regional nodal metastases and positive sentinel lymph node biopsy status. All primary tumors associated with co-existing distant metastases exhibited high PLK-1 expression in melanoma cells. Conversely, PLK-1 expression in stromal cells was more frequent in tumors without nodal metastases. PLK-1 expression in both compartments was not associated with survival.

Conclusion: PLK-1 expression is associated with metastatic potential in cutaneous melanoma.
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http://dx.doi.org/10.1111/cup.12985DOI Listing
October 2017