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R Zemer

Publications and source records attributed to R Zemer.

11 recordsLinked to original sources
PubMed2000

Detection of micrometastasis by cytokeratin-20 (reverse transcription polymerase chain reaction) in lymph nodes of patients with endometrial cancer.

BACKGROUND: Cytokeratins are constituents of the intermediate filaments (IFs) of epithelial cells which are expressed in various combinations, depending on the type of epithelium and degree of differentiation. We have reported (R. Zemer, A. Fishman, J. Bernheim, S. Zimlichman, O. Markowitz, M. Altaras, and A. Klein, Gynecol Oncol 70:410-413, 1998) on the determination of cytokeratin-20 (CK-20) by reverse transcription polymerase chain reaction (RT-PCR) in the detection of endometrial cancer cells as a potential biomarker. In that study, we also found that by using immunocytochemistry, most carcinomas were found to be negative for CK-20. The sensitivity and specificity rates obtained by using the RT-PCR method were 94.4 and 91%, respectively. OBJECTIVE: The aim of this study is to investigate the feasibility and potential of the specific mRNA marker, CK-20, to detect endometrial cancer cells-micrometastases (MMs)-by RT-PCR in lymph node (LN) samplings of patients undergoing hysterectomy for endometrial carcinoma. METHOD: We used the RT-PCR method to determine the expression of CK-20 in the LNs of 20 patients [study group (SG)] who were being surgically staged and treated for endometrial carcinoma. The specificity of the mRNA CK-20 marker was examined in LNs obtained from five healthy patients [control group (CG)] who underwent abdominal hysterectomy and bilateral salpingo-oopherectomy for benign gynecologic conditions. The LNs obtained from the SG and CG patients were prepared together before mRNA extraction. RNA of the various cell pellets was extracted and RT-PCR was performed with CK-20 primers. RT-PCR products were analyzed by agarose gel electrophoresis and ethidium bromide staining against PCR size markers. Specificity of the RT-PCR products was examined by Southern blotting. RESULTS: Histopathologic examinations demonstrated the presence of metastases in two (10%) SG patients. These patients were also CK-20 positive. Of the remaining 18 patients with negative histopathologic results, 6 (33%) were CK-20 positive and 12 (67%) were negative. All the CG patients were CK-20 negative (specificity, 100%). CONCLUSIONS: The results obtained in this study suggest that RT-PCR of CK-20 is more sensitive than traditional histopathologic methods in the diagnosis of MMs in LNs of patients with endometrial cancer. Thus, due to the aforementioned characteristics of CK-20, it may be considered a powerful biomarker in the detection of MMs in LNs of patients with endometrial cancer.

Adenocarcinoma↗
PubMed2000

Detection of tumor circulating cells by cytokeratin 20 in the blood of patients with endometrial carcinoma.

BACKGROUND: Cytokeratins are constituents of the intermediate filaments of epithelial cells which are expressed in various combinations depending on the epithelial type and the degree of differentiation. Using the reverse transcriptase-polymerase chain reaction technique (RT-PCR) we recently demonstrated that: (1) Cytokertin 20-the most recent discovered cytokeratin-is expressed in endometrial carcinoma tumors but not in the endometrium of patients with benign diseases, and (2) CK-20 is not expressed in blood cells. The aim of this study is to examine whether CK-20 expression in blood can be used as a biomarker for the detection of the dissemination of malignant cells in patients treated for endometrial carcinoma. METHODS: In the present study, we have used RT-PCR to determine the expression of CK-20 in the peripheral blood of the following groups: (1) preop new diagnosed patients (n = 20), (2) patients with no clinical evidence of disease following completion of definitive treatment (n = 33; 17 at low risk; 16 at high risk), (3) patients with recurrent disease (n = 6), and (4) a control group of healthy subjects (n = 16). RNA was extracted from cell pellets and analysed by RT-PCR using primers for CK-20. RESULTS: Of the 20 patients of the first group 7 (35%) were CK-20 positive. Of the 33 patients of the second group 17 (51%) were CK-20 positive. Subdivision of this group showed that 9 of 17 (53%) were positive in the low-risk subgroup, and 8 of 16 (50%) were positive in the high-risk subgroup. All 6 patients with recurrent disease were positive, and all subjects in the control group were negative. CONCLUSION: These results indicate that RT-PCR of CK-20, because of its high sensitivity, is a potential biomarker for detecting metastasis in blood samples of patients with endometrial carcinoma.

Biomarkers, Tumor↗
PubMed1999

Expression of cytokeratin 20 in the blood of patients with disseminated carcinoma of the pancreas, colon, stomach, and lung.

BACKGROUND: Cytokeratins are constituents of the intermediate filaments of epithelial cells that are expressed in various combinations depending on the epithelial type and the degree of differentiation. The recently identified cytokeratin 20 (CK-20) was found to be expressed in colonic, gastric, and pancreatic carcinoma tumor tissues. A low rate of incidence of expression of CK-20 was found in tumor tissue from lung carcinoma but no expression was found in blood even with the sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) method. The objective of the current study was to examine whether CK-20 expression in the blood can be used as a biomarker for the detection of dissemination in patients with carcinoma of the colon, stomach, and pancreas. METHODS: In the current study, RT-PCR was used to determine the expression of CK-20 in the blood cells from patients with metastatic colon carcinoma (n = 22), metastatic pancreatic carcinoma (n = 28), metastatic gastric carcinoma (n = 18), metastatic lung carcinoma (n = 13), no metastatic colon carcinoma (n = 13) and no known malignant diseases (n = 22). RNA was extracted from cell pellets and analyzed by RT-PCR using primers for CK-20. RESULTS: In the group of 22 patients with metastatic colon carcinoma, 14 were found to be CK-20 positive (sensitivity of 63.6% and specificity of 92.3%), 22 of the 28 pancreatic carcinoma patients showed positive CK-20 expression, and 12 of 18 patients with gastric carcinoma showed positive CK-20 expression. All patients with metastatic lung carcinoma except 1 were negative (12 of 13 patients), and 12 of 13 patients with colonic carcinoma with no known metastases also were negative. Negative CK-20 results were obtained in all 22 patients with no known malignant diseases. CONCLUSIONS: The results of the current study indicate that because of its high sensitivity, RT-PCR of CK-20 is a potential biomarker for detecting metastases in blood samples from patients with carcinoma of the colon, stomach, and pancreas.

Adenocarcinoma↗
PubMed1999

Telomerase activity in ductal carcinoma in situ of the breast.

Telomerase plays an important role in maintaining the stability of the chromosomes. Activity of telomerase has been detected in proliferating and immortalized cell lines and in a number of malignant tumors including invasive breast cancer. The aim of the study was to examine telomerase activity in ductal carcinoma in situ (DCIS), which is considered to be a precursor lesion of infiltrating breast carcinoma, using a PCR-based telomerase activity protocol (TRAP). We examined 35 samples obtained from histologically confirmed breast biopsies, including 13 normal breast tissues, 11 infiltrating ductal carcinoma (IDC), nine DCIS, and two DCIS with microinvasion. Telomerase activity was demonstrated in 8/9 samples of DCIS, both samples of DCIS with microinvasion, and all but one sample of IDC. Normal breast tissue had no demonstrable telomerase activity. Our results indicate that telomerase is activated frequently in early breast carcinogenesis, although its utilization as a biomarker in DCIS is questionable.

Adult↗
PubMed1998

Expression of cytokeratin 20 in urinary cytology of patients with bladder carcinoma.

BACKGROUND: Of the 20 known cytokeratins, CK-19 is expressed in normal urothelium, whereas the recently identified CK-20 is expressed in urothelial carcinoma cells but not in normal urothelial cells. The aim of this study was to examine whether CK-20 expression could serve as a noninvasive test in which malignant urothelial cells in urine are detected and monitored. METHODS: In the current study, the authors used reverse transcriptase-polymerase chain reaction (RT-PCR) methods to determine the expression of CK-20 in cells separated from the urine of patients with bladder carcinoma. Cells were obtained from the urine of 87 patients divided into the following 2 groups: 1) 14 healthy volunteers without any known history of transitional cell carcinoma (TCC), and 2) 73 patients with hematuria suspected for TCC of the bladder. For control purposes, CK-20 expression was examined in cells of 1) bladder carcinoma tumors of 5 patients, 2) blood of either patients with bladder carcinoma (n = 5) or healthy controls (n = 5), and 3) three different cell lines. RNA of the various cell pellets was extracted and RT-PCR was performed with CK-20 and CK-19 primers (CK-19 was used as a marker for normal epithelial cells). RESULTS: CK-20 amplification band (370 bp) was obtained with mRNA extracted from TCC cells of either bladder tumor or HT-29 line (a CK-20 colon carcinoma line). Sensitivity of the method was found to be 91%, whereas specificity was 67%. Among the 7 false-positive cases, 3 showed atypia, 3 hyperplasia, and 1 metaplasia, and 2 underwent previously successful TCC tumor removals, suggesting that the CK-20 test also responded to premalignant lesions. No false-positive cases were found in the healthy control group. No other preparation, including blood of the patients of with TCC, showed the CK-20 amplification band. CONCLUSIONS: These results indicate that CK-20 is a potential biomarker for noninvasive detection of bladder carcinoma by assaying uroepithelial cells from the voided urine specimen with RT-PCR.

Adult↗
PubMed1998

Expression of cytokeratin-20 in endometrial carcinoma.

OBJECTIVE: Cytokeratins are constituents of the intermediate filaments of epithelial cells which are expressed in various combinations depending on the epithelial type and the degree of differentiation. The recently identified cytokeratin-20 (CK-20) was found with immunohistochemical methods to be expressed in gastrointestinal epithelium, uroepithelial cells, and Merkel cells. Clues were also found for low expression of CK-20 in endometrial carcinoma cells. The aim of this study was to examine whether CK-20 expression can be measured in endometrial carcinoma with the more sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) methods and therefore used as a potential diagnostic tumor biomarker for endometrial carcinoma. METHODS: In the present study we have used RT-PCR methods to determine expression of CK-20 in endometrial epithelial cells. Endometrial specimens were collected from 18 patients with endometrial carcinoma and 22 patients that underwent hysterectomy due to benign diseases. The specimens were prepared for both RT-PCR and immunohistochemical analysis. RNA, of the various cell pellets, was extracted and RT-PCR was performed with CK-20 and CK-19 primers (CK-19 was used as a marker for normal epithelial cells). Immunohistochemistry was carried out with the avidin-biotin-peroxidase complex method on formalin-fixed paraffin sections using CK-20 antibody. RESULTS: CK-20 amplification band (370 bp) was obtained with mRNA extracted from endometrial carcinoma cells of 17 of the patients with endometrial carcinoma (sensitivity, 94.4%). CK-20 was negative in 21 patients with benign endometrial disease (specificity, 91.3%). No positive results were obtained with the immunohistochemical methods. CONCLUSION: These results indicate that RT-PCR of CK-20, because of its high sensitivity, is a potential biomarker for detecting endometrial carcinoma.

Adult↗
PubMed1998

Cytokeratin 20: a new marker for early detection of bladder cell carcinoma?

PURPOSE: Cytokeratins are constituents of the intermediate filaments of epithelial cells in which they are expressed in various combinations depending on epithelial type and degree of differentiation. Of the 20 known cytokeratins, cytokeratin 19 is expressed in normal urothelium cells, whereas the recently identified cytokeratin 20 (CK-20) is expressed in urothelial carcinoma but not normal urothelium cells. We examine whether CK-20 expression can be used as a bladder tumor marker for transitional cell carcinoma in cells isolated from urine. MATERIALS AND METHODS: The reverse transcriptase polymerase chain reaction method was used to determine expression of CK-20 in cells separated from urine of patients with bladder carcinoma. Cells were obtained from urine of 192 patients stratified into 3 groups of 21 healthy young volunteers without a history of transitional cell carcinoma, 27 with a negative bladder biopsy for transitional cell carcinoma and 144 with bladder transitional cell carcinoma. The parameters were tumor stage and grade, tumor size, number of tumors, urinary cytology and CK-20. RESULTS: CK-20 amplification band (370 base pairs) was obtained with messenger ribonucleic acid extracted from transitional cell carcinoma cells of bladder tumor. CK-20 in the urine samples of the control group was negative (no false-positive results, specificity 100%). Among the 27 patients with pTo disease CK-20 was negative in 20 (specificity 74.1%). In the 7 patients with positive CK-20 histology showed chronic inflammation in 2, atypical hyperplasia in 3 and metaplasia in 1. In 1 patient who had a known history of transitional cell carcinoma the urothelium was normal. Among 144 patients with bladder transitional cell carcinoma CK-20 was positive in 131. Sensitivity of the method was much higher than urinary cytology (91 versus 56.3%, p <0.0001). We demonstrated no correlation between CK-20 and tumor grade. CONCLUSIONS: Our results indicate that CK-20 is a potential marker for bladder cancer. The noninvasive detection method assesses urothelial cells from the voided urine specimen using reverse transcriptase-polymerase chain reaction. The CK-20 marker was significantly more sensitive than urinary cytology.

Adult↗
PubMed1997

Lymphoma with multi gene rearrangement on the level of immunoglobulin heavy chain, light chains, and T-cell receptor beta chain.

A unique case with diffuse mixed malignant lymphoma was investigated for gene rearrangement on the level of T-cell receptor (TCR), heavy chain immunoglobulin (Ig), and both light chains. Cell phenotype was examined with immunofluorescence techniques using antibodies against surface immunoglobulins (SIg) and the kappa and lambda light chains. Monoclonal antibodies were used against CD3, CD4, CD5, CD8, CD10, CD19, CD22, HLA-DR, and TdT. Gene rearrangement analysis for monoclonality determination was carried out with restricted DNA (EcoR I and Hind III) hybridized with one of the following 32P-labelled probes: T-cell receptor (TCR beta), immunoglobulin heavy chain (JH), k light chain, and lambda light chain. Phenotyping of the cell population from the excised lymph node (LN) revealed the presence of 66% B-cells and 35% T-cells. Most of the B cells (94%) expressed mu heavy chain only. Expression of both light chains was negligible (k = 7% and lambda = 2%). Gene rearrangement, which indicates monoclonality, was positive on the level of TCR, Ig heavy chain, and both light chains. The data obtained suggests a neoplastic transforming event in lymphoid stem cells, which preceded the subsequent differentiation process into either B or T lymphoma.

Aged↗
PubMed1997

Induction of apoptosis by chlorodeoxyadenosine and dexamethasone in B-chronic lymphocytic leukemia.

Apoptosis, programmed cell death, occurs in a variety of cellular systems and in response to many different stimuli. In the present study we examined the ability of dexamethasone (Dex) and chlorodeoxyadenosine (2-CdA) to induce apoptosis in lymphocytes of patients with B-chronic lymphocytic leukemia (B-CLL). Lymphocytes of 29 untreated patients and 9 healthy controls were isolated and incubated for 24 hours in the presence or absence of either Dex (2 microM) (n = 15) or 2-CdA (3 microM) (n = 14). Following incubation the cells were harvested and their DNA extracted and analysed for internucleosomal DNA cleavage by UV illumination after electrophoresis on agarose slab gel containing ethidium bromide. In the Dex group, 10 patients showed dexamethasone independent spontaneous apoptosis appearing 24 hours after the start of incubation. These were the only instances of dexamethasone-enhanced apoptosis. Five patients showed no spontaneous or dexamethasone induced apopto sis. Of the 2-CdA group, 5 showed spontaneous apoptosis enhanced by 2-CdA. No spontaneous apoptosis was observed in the cells from 9 other patients, however, 2-CdA induced apoptosis in 8 cases in this group. This study shows that monitoring of apoptosis in CLL may provide important information regarding susceptibility of the cells to drug induced apoptosis.

Antineoplastic Agents, Hormonal↗
PubMed1994

B-cell malignancy after low grade T-cell lymphoma.

BACKGROUND: Low grade, small lymphocytic, non-Hodgkin's lymphoma, was diagnosed in a 38-year-old woman. Thirty months after the initial diagnosis was made, a population of lymphoid cells with pathologic morphology was found in the patient's peripheral blood (PB). Cell phenotyping was performed and monoclonality was analyzed in cells obtained from a removed lymph node (LN) and the PB of the patient. METHODS: The cell phenotype was examined with immunofluorescence techniques using antibodies against SIg and monoclonal antibodies against CD1, CD3, CD4, CD5, CD8, CD19, and the kappa, and lambda light chains. Gene rearrangement analysis for monoclonality determination was performed with restricted DNA (EcoRI, Hin-dIII and BamHI) hybridized with either 32P-labeled T-cell receptor DNA probe (TcR-beta) or immunoglobulin-heavy chain probe (JH). RESULTS: With regard to the cell population of the removed LN, cell phenotyping showed the predominance of CD4+ T-cells over a polyclonal B-cell population. Gene rearrangement analysis proved the monoclonal nature of the T-cells and the polyclonal nature of the B-cells. As to the PB, gene rearrangement and cell phenotyping of the lymphocytes showed the predominance of monoclonal kappa type B-cells over polyclonal T-cells. CONCLUSIONS: The data obtained suggest two unrelated lymphoproliferative diseases in this patient, expressed as monoclonal T-cell population in LN and as monoclonal B-cell population in PB.

Adult↗