role of RCC, vimentin and cytokeratin 7 in the differential diagnosis of conventional renal cell carcinoma

The present study investigates the value of RCC, Cytokeratin 7 and vimentin immunohistochemical staining in the distinction between granular variant of renal cell carcinoma, chromophobe renal cell carcinoma and oncocytoma. Chromophobe renal cell carcinomas were characterized by positive cytoplasmic immunoreactivity to CK7 [100%] while they were negative to vimentin, and RCC. Conventional renal cell carcinoma showed positive immunoreactivity to; vimentin [100%], RCC [73%], and CK7 [14%]. On the other hand, oncocytoma showed focal weak staining to RCC while they were negative to the other markers. A panel of antibodies including RCC, Cytokeratin 7, and vimentin is useful tool in differentiating chromophobe renal cell carcinoma from granular type renal cell carcinoma and oncocytoma

Multilocular cystic renal cell carcinoma: clinicopathological and immunohistochemical characters

Multilocular cystic renal cell carcinoma [MCRCC] is a rare and distinct subtype of renal cell carcinoma [RCC]. It is characterized by unique gross and microscopic features. As well as the outcome of this tumor is excellent in comparison to the conventional type of RCC. Additionally, there is no evidence of progression or recurrence is seen after surgery among MCRCC, differing from conventional RCC. The aim of this work is to evaluate the clinicopathological features of this tumor. Also, to study their immunohistochemical characters comparing with conventional RCC. As well as try to see the differential diagnosis between MCRCC and other renal cystic lesions in particular RCC with cystic areas, cystic nephroma, and multilocular renal cysts. This work was performed on 20 cases of different renal tumors. These cases were collected retrospectively from the surgical pathology files at Department of Pathology, King AbdulAziz University Hospital, Jeddah through the period 2000 to 2007. The clinicopathological parameters including gross and microscopic features were studied. Then panel of immunohistochemical markers including epithelial membrane antigen [EMA], cytokeratin pan [CK-pan], renal cell carcinoma marker [RCC], CD10, vimentin, and cytokeratin 7 [CK7] were preformed. Other additional markers were added when needed. According to the criteria of the World Health Organization [WHO] 2004, the properly selected cases with full criteria of MCRCC were detected in 4 cases [20%]. The remaining of the studied cases were 13 cases [65%] conventional type RCC, one case [5%] renal leiomyosarcoma, one case [5%] sarcomatoid RCC and one case [5%] was clear cell renal cell sarcoma. The age of all cases was ranged from 2 years to 60 years [the mean was 50 years]. The gender of the included cases was 14 cases [70%] male and 6 cases [30%] were female. Yet, the gender of the included MCRCC cases in our study was 3 cases [75%] male and one case [25%] was female. Their ages were ranged from 30 years to 60 years. 3 cases [75%] of MCRCC were Fuhrman grade 1 and one case was Fuhrman grade2. As well as 3 cases of them were suffered from loin pain and in one case the tumor was discovered incidentally during routine abdominal ultrasound check-up. All cases were undergone for nephrectomy. Histologically there were variable sized cysts lined by single layer of neoplastic cells. The tumor stage in all cases of MCRCC was 1. So, MCRCC has a lower pathologic nuclear grade and stage. Immunohistochemically MCRCC cases were immunoreactive for EMA, RCC marker, vimentin, and CK7. Yet, MCRCC cases showed lowered immunoreactivity for CD10, and all eases were lacked cytokeratin 20 [CK20]. MCRCC represents a distinct subtype of RCC. The prognosis of MCRCC is excellent in comparison to other types of RCC. Most of MCRCC cases were Fuhrman grade 1, and stage 1. Some cases of MCRCC can be seen incidentally during investigation for unrelated conditions. The diagnosis of MCRCC is strictly related to its unique gross and microscopic features. As well as this tumor must be differentiated from other cystic renal lesions. Immunohistochemically these tumors were immunoreactive for the same markers of conventional RCC. Yet, CD10 showed lowered percentage of positive immunoreactivity, and CK7 showed strong positivity in comparison to conventional RCC

Expression of CD44 protein and cytokeratin 20 in non Bilharzial and Bilharzial bladder carcinoma

Carcinoma of the urinary bladder is one of the most common cancers world wide. It is the fourth most common malignancy in males and the ninth most common malignancy in females. CD44 is a family of cell-surface transmembrane glycoproteins that serve as receptors for hyaluronate and bind extracellular matrix components. CD44 plays a definite role in cell- cell and cell- matrix interactions. Thus, its down-regulation would facilitate loss of cell - cell cohesion, detachment from the basement membrane, and subsequent infiltration of the underlying tissues. It is suggested that the expression of CD44 is associated with differentiation and prognosis in bladder carcinoma. Cytokeratins [CKs] are a family of proteins that form the intermediate filament cytoskeleton of epithelial cells. Cytokeratin 20 [CK20] has been proposed as a marker of neoplastic change as well as a predictor for progression of urothelial carcinoma. The aim of the work is to study the immunohistochemical expression of CD44 and CK20 in carcinoma of the urinary bladder and correlate the immunohistochemical expression of CD44 and CK20 with different prognostic parameters including the grade and stage of the studied tumors. The studied 71 specimen were subjected to the ordinary H and E staining and immunohistochemical staining for CD44 and CK20. Correlative studies between CD44 and CK20 expression with different prognostic parameters including the grade and stage of the studied tumors revealed statistically significant correlation between CD44 and CK20 expression and the tumor grade and stage of urothelial carcinoma cases. No relation was found between the expression of CD44 and CK20 and the presence of Bilharziasis. Loss or reduction of CD44 immunoreactivity and increasing CK20 positiviiy were significantly associated with increasing tumor grade and stage in the studied cases of urothelial carcinoma and to each other, so, they have combined utility in predicting the behaviour and prognosis of urothelial carcinoma, with no significant difference in their expression between non Bilharzial and Bilharzial bladder carcinomas

Avaliação dos valores sérico e pleural dos marcadores tumorais CEA, CYFRA21-1 e CA 15-3 em portadores de derrame pleural / Evaluation of serum and pleural levels of the tumor markers CEA, CYFRA21-1 and CA 15-3 in patients with pleural effusion

OBJETIVO: Dosar os marcadores tumorais antígeno carcinoembrionário (CEA), fragmento da citoqueratina 19 (CYFRA21-1) e antígeno glicosídico associado a tumor 15-3 (CA 15-3) em sangue e líquido pleural de portadores de derrames pleurais benignos e malignos, avaliando a sensibilidade de cada um deles nesses fluidos. MÉTODOS: Avaliamos prospectivamente 85 pacientes com derrame pleural. O estudo do líquido pleural obedeceu a critérios determinados pela literatura. A dosagem dos marcadores foi realizada por eletroquimioluminescência. A sensibilidade foi determinada sob a condição de que a especificidade fosse > 90 por cento. RESULTADOS: Foram diagnosticados 36 casos malignos (42,4 por cento), 30 benignos (35,3 por cento); em 19 pacientes (22,3 por cento), o diagnóstico foi inconclusivo. Nos casos malignos, os valores de CEA e CYFRA21-1 foram maiores no líquido pleural do que no sangue, fato não observado para o CA 15-3. Nos casos benignos, os valores do CYFRA21-1 foram maiores no líquido pleural do que no soro, enquanto que para o CEA e o CA 15-3, ocorreu o oposto. Todos os marcadores apresentaram diferença significativa entre os casos malignos e benignos, em líquido pleural e soro. Foi encontrada sensibilidade para CEA, CYFRA21-1 e CA 15-3 no líquido pleural de 69,4 por cento, 69,4 por cento e 66,7 por cento, respectivamente e quando associados, foi 80,6 por cento. No soro, a sensibilidade foi 57,1, 71,4 e 48,6 por cento para CEA, CYFRA21-1 e CA 15-3, respectivamente, e quando associados, foi 77 por cento. CONCLUSÃO: Os resultados sugerem que a utilização desses marcadores pode ser útil na diferenciação entre derrames pleurais malignos e benignos.

OBJECTIVE: To determine the levels of the tumor markers carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21-1) and carbohydrate antigen 15-3 (CA 15-3) in the blood and pleural fluid of patients with benign or malignant pleural effusion, evaluating the sensitivity of each marker in these fluids. METHODS: We prospectively evaluated 85 patients with pleural effusion. The study of the pleural fluid observed the criteria established in the literature. Levels of the markers were determined using electrochemiluminescence. The sensitivity was determined on the condition that the specificity was > 90 percent. RESULTS: Of the 85 cases, 36 (42.4 percent) were malignant, 30 (35.3 percent) were benign, and the results were inconclusive in 19 (22.3 percent). In the malignant cases, the CEA and CYFRA21-1 levels were higher in the pleural fluid than in the blood, which was not observed for CA 15-3. In the benign cases, the CYFRA21-1 levels were higher in the pleural fluid than in the blood, whereas the opposite was found for CEA and CA 15-3. There were significant differences between malignant and benign cases for all markers, in pleural fluid and blood. In the pleural fluid, the sensitivity of CEA, CYFRA21-1 and CA 15-3 was 69.4, 69.4 and 66.7 percent, respectively, and the combined sensitivity was 80.6 percent. In the blood, the sensitivity was 57.1 percent, 71.4 percent and 48.6 percent for CEA, CYFRA21-1 and CA 15-3, respectively, and the combined sensitivity was 77 percent. CONCLUSION: The results suggest that these markers might be useful in the differentiation between malignant and benign pleural effusion.

Cytokeratin 20: a new promising marker for colorectal carcinomas

Colorectal carcinoma [CRC] is one of the most common malignancies. Inspire of successful resection of the primary Tumour, metastasis is one of the most important factors determining the patient's prognosis. The present study was initiated to detect disseminated cancer cells in LNs and peripheral blood of patient's with CRC via detection of cytokeratin 20 [CK20] mRNA expression by conventional and real time reverse transcriptase polymerase chain reaction [RT-PCR]. This study included 34 patients with early stage CRC who have undergone total tumor resection, and were proved to be node negative histopathology. The study also included two control groups; a pathological control group [n = 10] suffering from ulcerative colitis and a healthy control group [n = 21], who have undergone pelvic repair for rectal prolapse and were otherwise completely free. For all subjects, serum CEA was measured in addition to cull routine laboratory investigations. Conventional PCR for detection of CK20 mRNA was [lone in both peripheral blood and lymph node samples for all participants. Whereas, quantitative real time RT-PCR for detection of CK20 mRNA was done for lymph node samples of the patient group only. Results of' the present study showed that all peripheral blood samples tested by conventional PCR were consistently negative for CK20 mRNA expression, whereas, mRNA was detected in the LN tissues of 16 of the 34 patients included in this study. Quantitative real-time RT-PCR appeared to be more sensitive than conventional PCR as it was able to detect CK20 mRNA in 20 out of the 34 patients included in this study. Based on the results of our realtime PCR, time patient group was divided into 20 patients with detectable mRNA and 14 with undetectable CK20 mRNA. The results of CEA levels were elevated in the former goup than time latter one, however this elevation was not statistically significant. on comparing nodal expression of CK20 mRNA in different histologic tumour types, 85. 7% of mucinous carcinoma patients showed nodal expression versus 51.9% of adenocarcinoma patients. Patients with Dukes' stag B2 showed 92.8% nodal expression versus only 35% patients with Dukes' B1 stage. Poorly differentiated tumours showed 100% nodal expression of CK20 mRNA versus 38.1% for time moderately differentiated and 66.7% for the well differentiated tumour group. In conclusion, CK20 mRNA is a valuable tool for monitoring early stage dissemination of CRC malignant cells. Real time PCR provides a more sensitive and reliable technique for detection of' disseminated cancer cells in CRC patients than the traditional PCR technique. Besides, CK20 mRNA is a promising predictor of poor prognosis being increased in mucinous, poorly differentiated and Dukes' B2 CRC patients

Can immunohistochemistry differentiate between cells and malignant epithelial cells?

Background: Malignant epithelial cells can be difficult to distinguish from reactive benign mesothelial cells in cytological smears prepared from body fluid effusions. Calretinin and cytokeratin 5/6 are valuable markers in the immunohistochemical distinction between malignant mesothelioma and adenocarcinoma in tissue sections. However, there is limited and conflicting data regarding their utility in the differential diagnosis between reactive benign mesothelial cells and malignant cells in cytologic material

Aim: The aim of the current work is to evaluate the efficiency of IHC staining of calretinin, CK5/6, CK7, CK20 and TTF-1 in the distinction between reactive mesothelial cells and metastatic adenocarcinoma and the prediction of origin of primary carcinoma in cell blocks prepared from ascitic and pleural effusions

Material and Methods: Formalin-fixed paraffin-embedded cell block preparations of 20 cytologic fluids were stained immunohistochemically for monoclonal antibodies to cairetinin, CK5/6, CK7, CK20, TTF-1 and CD68

Results: Mesothelial cells showed positive immunoreactivity to calrelinin and CK5/6 [100%], and CD68 [30%] while they were totally negativei to CK7, CK20, and TTF-1 is useful contrast, malignant epithelial cells [in all studied cell block preparations] were negative to cairetinin, CK5/6 and CD68 while 50% were immunopositive to CK7, 34% to CK 20, and 15% to TTF-1

Conclusions: Cell block preparation is an excellent method for immunohistochemical staining of cytologic specimens. Calretinin and CK5/6 are valuable markers in the identification of reactive mesothelial cells. The use of an immunohistochemical panel of antibodies to calretinin, CK516, CK7, CK20, and TTF-1 is useful in the differential diagnosis between reactive mesothelial cells and metastatic adenocarcinoma in cytologic cell block preparations

Serum and biliary cytokeratin fragment CYFRA 21-1, as a marker of resectability for extrahepatic cholangiocarcinoma

Serum levels of CEA and CA 19.9 are occasionally elevated in cases of cholangiocarcinoma [CC]. They, however, have low sensitivity and specificity, which makes the diagnosis of CC difficult, and better tumor markers are needed. CYFRA 21-1, a cytokeratin fragment, is being evaluated as a tumor marker with diagnostic potential for small cell lung carcinoma. The resectability of CC is difficult to assess accurately pre-operatively using available laboratory and imaging techniques. The aim of the present study was to assess the diagnostic value of CYFRA 21-1, CA 19.9, and CEA in serum and bile of patients with extrahepatic CC. and to evaluate whether any of these markers can be used to assess resectabilily. Fifty patients with extrahepatic obstructive jaundice were included in this study. Group 1: 30 patients with histologically proven CC had serum and bile samples measured for CYFRA 21-1, CA 19.9; and CEA. Group II: 20 patients with calcular obstructive jaundice had serum and bile samples obtained during endoscopic removal of stones to control for the effect of biliary obstruction on CYFRA 21-1, CA 19.9, and CEA. Sixteen patients in whom imaging studies showed resectability were explored and operative findings were correlated to tumor markers. CEA and CA 19.9 were measured using enzyme immnunoassay [EIA,], and CYFRA 21-1 level was measured using electrochemiluminescence immunoassay [ECLIA] in serum and bile of the CC patients and the caicular obstructive jaundice controls. Patients with CC had significantly higher serum levels of CEA [24.5 +/- 6 vs 4 +/- 1.3 ng/ml. p<0.01] CA 19.9 [118.9 +/- 25.4 vs 31.2 +/- 8.4 U/ml, p< 0.01] and CYFRA 21-1 [8.3 +/- 2.7 vs 2.1 +/- 0.5 ng/ml, p<0.01] than patients with benign calcular cholestatic disorder, and bile concentration of the 3 markers was sign higher than serum concentrations. Among the 16 patients who underwent surgery, serum and biliary levels of CYFRA 21-1 were significantly higher with irresectable tumors. CYFRA 21-1 may be a useful marker for resectabilily saving many unnecessary operations. This, however, needs further larger studies to be documented

Expression of intercellular adhesion molecule ICAM-1 and cytokeratin as Potential markcrs of disease activity in the Egyptian pattern of chronic hepatitis C

Chronic hepatitis C [ch.HCV] and schistosomal hepatic fibrosis or both as a mixed hepatic lesion [MHL] are among the most common causes of endemic chronic hepatic disease in Egypt. Adhesion molecules especially ICAM-1 play an important role in inflammatory and immunological responses of chronic liver disease. Cytokeratin 18 [CK-18], although normally expressed in hepatic tissue, yet it is altered during chronic inflammatory hepatic lesions. This work was planned to study ICAM-1 as expressed in hepatic tissue in the different grades of the disease activity, in relation to its circulating levels in patients sera, and to evaluate the level of CK-18 expression in relation to the different grades of chronic inflammation and stages of fibrosis in the examined liver biopsies. The material for this study comprised 33 patients [17 ch.HCV and 16 MHL]. Seven cases, that proved to have nearly normal serological data and insignificant histopathological hepatic features, were considered as controls. All patients were assessed for HCV serological markers as well as serum levels of soluble ICAM-1 [sICAM-1] by the Enzyme Linked Immunosorbent. Assay [ELISA]. Liver needle biopsy specimens were processed and assessed for the histopathological grade of the disease activity and stage of fibrosis of the hepatic lesion. Tissue expression of ICAM-1 and CK-18 was detected using immunohistochemical techniques. Our results revealed significantly higher levels of serum sICAM-1 in both ch.HCV and MHL groups compared to controls [p<0.01]. Meanwhile, higher levels of sICAM-1 were recorded in the MHL cases relative to ch.HCV cases. ICAM-1 expression was not detected in any of the control cases, while it was positively expressed in all ch. HCV and MHL cases, with a higher score recorded in the later group [P>0.001 compared to the control group]. ICAM-1 expression was detected mainly within the sinusoidal cells [endothelial and Kupffer cells], hepatocytes, mononuclear inflammatory cells and vascular endothehail cells in portal areas. On classifying patients according to their grades of active inflammation and stages of fibrosis, higher scores of ICAM-1 expression within the hepatic tissue were recorded in cases with more active inflammatory grades and higher fibrotic stages. On the other hand, serum of ICAM-1 levels though were significantly elevated in patients with higher grades of inflammatory activity yet, they were decreased in patients with higher fibrotic stages. CK18 expression was mainly detected within hepatocytes of the periportal areas [in a combined membranous and intracytoplasmic pattern], as well as within the bile ducts epithelium in the portal areas. Over expression of CKI 8 was detected in both the ch.HCV and MHL groups [p<0.001 relative to controls], but the expression scores were higher in the MHL group. From these results we may conclude that MHL is a more aggressive and active chronic inflammatory hepatic disease than ch.HCV alone. Also, serum levels of sICAM-1 as well as hepatic expression of both ICAM-1 and CK-18 are related to the degree of disease activity, which may point out to the possibility of using serum sICAM-1 levels as well as the expression scores of hepalic ICAM-1 and CK-18 as efficient tools for monitoring the disease activity in ch. HCV and MHL patients. While ICAM-1 expression in tissue could be used as indicator for the stage of fibrosis in those patients also recommend that both ICAM-1 in serum andhepatic tissue could be used for monitoring the effect of therapy on the studied pattern of chronic hepatitis C

Detection of K19 mRNA compared with tumour markers in peripheral blood from breast cancer patients

A rise in peripheral blood of the glycoproteins carcino-embryonic antigen [CEA] and CA15.3 may be indicative of a subsequent clinical relapse and provides a useful adjunct in the management of metastatic breast cancer. We wished to study the relationship of these tumour markers with the expression of keratin 19 [K19] mRNA as a marker of epithelial cancer cells in periodic blood samples taken from patients undergoing postoperative therapy. CEA and CA15.3 were measured by radio-immunoassay while K19 mRNA was assessed by gel electrophoretic separation after a 40-cycle polymerase chain reaction amplification. Analysis of 395 samples [regardless of patient] showed concurrence in the detection of K19 and CEA and CA15.3 [using positivity cut-offs of 3.2 ng/ml and 32 U/l, respectively] in <20% of cases that were K19+ and in >80% for samples that were K19-. The frequency of CEA and CA15.3 positivity was related to the amount of K19 product. For 65 patients with 2-3 samples collected at 6 monthly intervals, we observed complete agreement for detection of these markers [at each occasion] in about 30% of cases only. Conclusions: These discrepancies may be due partly to the currently qualitative nature of the K19 assessment and emphasize the need for quantification of this but may also reflect a difference in the source of the signals, i.e. K19 originating from circulating cells and CEA/CA15.3 shed from solid tumour deposits. It is likely that K19 detection is more sensitive, but it is less certain which of these markers reflects a significant systemic tumour load and hence is of greater value in predicting relapse

Utility of serum cytokeratin 19 fragment (CYFRA 21-1) and carcinoembryonic antigen (CEA) as tumour markers for non-small cell lung cancer.

Serum cytokeratin 19 fragment (CYFRA 21-1) and carcinoembryonic antigen (CEA) levels were determined with an enzyme immunoassay in 51 patients with non-small cell lung cancer (NSCLC), 26 patients with benign lung diseases and 26 normal individuals in order to evaluate their clinical utility in the diagnosis of NSCLC. Patients with NSCLC demonstrated higher serum CYFRA 21-1 and CEA levels than both patients with benign lung diseases and normal group. We used the cut off value which was derived from the 95th percentile value of CYFRA 21-1 and CEA levels in the group of patients with benign lung diseases; CYFRA 21-1 at 3.13 ng/ml and CEA at 7.7 ng/ml. The sensitivity and diagnostic accuracy of CYFRA 21-1 and CEA for the group of NSCLC patients were 66.7 per cent, 76.6 per cent and 35.3 per cent, 55.8 per cent, respectively. When combining CYFRA 21-1 with CEA, the sensitivity and diagnostic accuracy were 68.6 per cent and 66 per cent. These results suggest that CYFRA 21-1 and CEA are useful serum markers for the diagnosis of NSCLC; especially subtype squamous cell and adenocarcinoma, respectively. The usefulness is not enhanced by combining the assay of CYFRA 21-1 and CEA.

Análise em pacientes do hospital A. C. Camargo de dois marcadores tumorais: CA-72.4 e CYFRA-21.1 / Analysis in patients of the Hospital A. C. Camargo of two tumor markers: CA-72.4 and CYFRA-21.1

O comportamento dos marcadores CA-72,4 e CYFRA-21.1 foi analisado em pacientes do Hospital a. C. Camargo. A dosagem dos níveis séricos de CA-72,4 foi realizada em 49 pacientes por ensaio automatizado de quimioluminescência e mostrou-se útil nos dois grupos de pacientes analisados, com tumores de esôfago e gástricos. Em ambos os casos, houve um aumento de sensibilidade quando se acrescentou a dosagem deste marcador à do antígeno carcinoembriônico (CEA) ou ao marcador CA-19.9. Foram mais sensíveis as associaçöes CA-72.4 e CA-19.9 para os tumores de esôfago, e CA-72.4 e CEA para tumores gástricos. Já o marcador CYFRA-21.1 foi analisado por radioimunoensaio em 102 doadores normais do banco de sangue do HACC e em 20 pacientes com tumores de pulmäo. Nestes últimos, o CYFRA-21.1 esteve alterado em 55 por cento dos casos. Desta forma, o marcador CYFRA-21.1 pode ser considerado, como a enolase neurônio-específica (NSE), uma ferramenta muito útil no seguimento de pacientes com tumores de pulmäo

Cytokeratin 20 [CK-20] a prognostic marker in TCC of the bladder

This immunohistochemical study has examined the degree and pattern of expression of a recently identified cytokeratin [CK-20] in 10 cases of normal bladder urothelium and 15 cases of TCC with well documented clinical course, using immunoperoxidase technique. In normal urothelium the expression of CK-20 was found to be differentiation related and confined to the superficial highly differentiated "umbrella" cells. Correlation of the expression of CK-20 with the clinical course in TCC of the bladder revealed that 85.71 of superficial bladder tumour with good outcome had retained a high degree of expression of CK-20: in the surface cell layer, whereas 80% of muscle invasive tumours, with aggressive clinical course, showed negative expression of the antigen. These observations suggest that the degree and pattern of expression of CK-20 is parallel to the outcome of the tunour. which may make CK-20 a useful prognositc marker in TCC of the bladder. There was another group which showed heterogenous pattern of expression of CK-20. This group was represented mainly by 3 cases of T1 tunours and who had one or more recurrences during the period of follow-up