Proliferating Cell Nuclear Antigen Grade Research Articles

Chromoendoscopy with hematoxylin in the classificaton of gastric lesions

Hematoxylin is a basic histological stain which stains nuclei. Apparently, it can be used with endoscopy to diagnose malignant lesions due to their increased nucleus-tocytoplasm ratio. The aim of this study was to evaluate the use of hematoxylin in the differentiation of gastric lesions. Part 1. In a prospective study, a total of 70 different gastric abnormalities were stained with 1% hematoxylin solution during upper endoscopy. Results of histological examinations were compared with endoscopic findings. Part 2. During a 5-day period, antral mucosa was stained with hematoxylin in all patients (n = 26), independent of the upper endoscopy results. Grades of stained mucosa were compared with the proliferating cell nuclear antigen (PCNA) index, which was examined using an immunohistochemical method. Part 3. Gastric biopsies (total, 12) were obtained from the stained and unstained areas within the limits of one lesion (total, 6) after chromoscopy with hematoxylin and examined with monoclonal (PCNA) antibodies. Using histological findings, the sensitivity and specificity of chromoendoscopy with hematoxylin for detecting gastric neoplasia and adenocarcinoma were determined. A significant positive correlation (r = 0.81; P < 0.001) was found between the PCNA index and grade of endoscopic stained mucosa. The PCNA index was increased in stained areas of gastric abnormalities in comparison to unstained areas within the limits of one lesion in all six patients. Chromoendoscopy with hematoxylin increases opportunities for precision biopsy. The grade of endoscopic stained mucosa depends on epithelial cell proliferation and is associated with malignancy. By using histology as the main outcome, the sensitivity and specificity of chromoendoscopy with hematoxylin for diffuse staining in the detection of gastric neoplasia and adenocarcinoma were 92.9% and 89.3%, respectively.

Proliferating cell nuclear antigen expression in papillary thyroid carcinoma.

To correlate the expression of proliferating cell nuclear antigen (PCNA) with various clinicopathological features of papillary thyroid carcinoma. Sections from 131 formalin fixed, paraffin wax embedded papillary thyroid carcinomas were stained with a monoclonal antibody (PC10) directed against PCNA using the avidin-biotin immunoperoxidase (ABC) method. PCNA immunoreactivity was based on the PCNA labelling index (LI) following evaluation of at least 1000 tumour cells, and expressed as follows: grade A (LI < 10%), grade B (10% < or = LI < 25%), and grade C (LI > or = 25%). The relation between PCNA expression in these three groups and other clinicopathological factors, such as sex, age, tumour size, nodal metastases, and histological differentiation, were examined. Based on the labelling index, 57 (43.5%) cases were graded as A, 46 (35.1%) as B, and 28 (21.4%) as C. The female-:male ratios were 6.13:1 for group A, 2.83:1 for group B, and 2.11:1 for group C. The mean (SD) ages of the patients were 39.0 (16.1) in group A, 53.5 (14.4) in group B, and 55.8 (13.3) years in group C. The correlation between age and PCNA grade was strongest in women. PCNA immunoreactivity is correlated with sex and age in patients with papillary thyroid tumours.

Immunohistochemical detection of proliferating cell nuclear antigen in middle ear cholesteatoma.

Cholesteatoma epithelium is characterized by a keratinocyte dysregulation accompanied by destruction of the ossicles and temporal bone. Immunohistochemical methods using antibodies to cell-cycle-related antigens can be used as a means for assessing various aspects of proliferation in cholesteatoma tissue. They also have the important advantage of preserving the spatial orientation of proliferating cells in histological sections. Proliferating cell nuclear antigen (PCNA) is a 36 kDa DNA-delta-polymerase-associated protein that is directly involved in the mechanisms of DNA synthesis. In the present study the expression of PCNA was investigated in formalin-fixed, paraffin-embedded biopsy specimens of cholesteatomas and normal skin. Normal skin revealed nuclear staining in a small number of keratinocytes (PCNA grade, 1.5) located in the basal cell layer. In contrast, an increased number of PCNA-labeled basal and suprabasal epidermal cells (PCNA grade, 9.3) were found in cholesteatoma samples. Our findings indicate that PCNA represents a reliable marker for epithelial proliferation, showing that cholesteatoma epithelium proliferates at a higher rate than normal epidermis. These findings also support the concept of keratinocyte dysregulation in middle ear cholesteatoma.

Expression of proliferating cell nuclear antigen in premalignant lesions of the larynx

Purpose: This study was performed to 1) evaluate the proliferative activity in premalignancies of the larynx by immunostaining for proliferating cell nuclear antigen (PCNA), 2) to examine the relationship between PCNA expression and histological diagnosis and 3) to evaluate clinical values of PCNA immunostaining. Materials and Methods: The percentage of PCNA-positive nuclei (PCNA index) and their highest location (PCNA grade) were evaluated in 44 specimens of premalignant lesions, 7 premalignant lesions near carcinomas, 9 normal epithelium, and 12 early invasive carcinomas of the larynx. Follow-up was conducted. Results: The mean of PCNA indices in premalignancies was significantly higher than in normal epithelium but lower than in early invasive carcinomas. PCNA index had no apparent correlation with the histological diagnosis, but in the same case, the higher degree of dysplasia showed higher PCNA index. PCNA grade showed significant correlation with the histological diagnosis. Cases with higher PCNA expression showed higher percentages of recurrence and cancerization. Premalignant lesions near carcinomas presented significantly high PCNA expression. Conclusions: The proliferative activity in premalignancies of the larynx is higher than in normal epithelium and lower than in carcinomas. It differs from case to case, but in the same case the higher degree of dysplasia possesses higher proliferative activity. Dysplasia is associated with abnormal proliferation pattern. Special attention should be paid to those with abnormal proliferation patterns as well as high proliferative activity. PCNA immunostaining is of some referential value in predicting clinical outcome and guiding to make an early diagnosis of malignancy.

EXPRESSION OF EPIDERMAL GROWTH-FACTOR (EGF), MATRIX METALLOPROTEINASE-9 (MMP-9) AND PROLIFERATING CELL NUCLEAR ANTIGEN (PCNA) IN ESOPHAGEAL CANCER

Expression of human epidermal growth factor (EGF) and matrix metalloproteinase-9 (MMP-9/gelatinase B) was examined immunohistochemically in 62 cases of surgically resected esophageal carcinomas, and the correlation between EGF expression and the proliferative activity of the tumors was studied by analysing the number of proliferating cell nuclear antigen (PCNA)-positive cells. Expression of EGF and MMP-9 was observed in 16 (38.1%) and 18 (42.9%) of the 42 superficial carcinomas and 8 (40%) and 14 (70%) of the 20 advanced carcinomas, respectively. The differences in the MMP-9 expression between the superficial carcinomas and the advanced carcinomas was significant (p<0.05). The synchronous expression of EGF and MMP-9 was observed in 15 (24.2%) of 62 carcinomas, i.e. 62.5% of the 24 EGF-positive tumors expressed MMP-9, but there was no statistically significant correlation between the expression of EGF and MMP-9. The relationships between EGF expression and tumor proliferative activity and prognostic factors were investigated. The PCNA grades were significantly higher in tumors with EGF-positive than those with EGF-negative expression (p<0.05) and the EGF expression showed a good correlation between the expression of MMP-9 and vascular invasion (p<0.01). The expression of MMP-9 was stronger in the advanced than the superficial carcinomas and there was a good correlation with vascular invasion (p<0.01). In a follow-up study of 55 patients, those with tumor that expressed MMP-9 or had a high PCNA grade showed a poor prognosis. Taken together, these observations suggest that both EGF and MMP-9 participate in the invasive phenotype in human esophageal carcinoma, but the expression of EGF is not directly related to the expression of MMP-9. Additional growth factors and cytokines may be involved in regulation of MMP-9 expression in this carcinoma.

Proliferating cell nuclear antigen (PCNA) in high-grade malignant fibrous histiocytoma: prognostic value in 48 patients.

Grading based on histopathologic features is used to predict survival in soft-tissue sarcoma. However, variations in clinical behavior between tumors of the same grade motivate a search for additional factors that correlate with prognosis. Proliferating cell nuclear antigen (PCNA) is expressed in proliferating cells during the G1, S and G2-phases. To evaluate a prognostic implication of PCNA, the tumors of 48 patients with malignant fibrous histiocytomas (13 grade III, 35 grade IV) with a minimum follow-up of 2 years were immunohistochemically studied. We used PC10, a monoclonal antibody (MAb) directed against PCNA, which allows cell proliferation in formalin-fixed, paraffin-embedded tumor tissue to be evaluated. We applied a semiquantitative PCNA grading scheme to all stained nuclei of an entire slide. The 3-year metastasis-free survival rate was 0.87 for patients in grade A (low PLNA rate), and 0.14 for patients in grade C (high PLNA). Our findings show that immunohistochemical evaluation of cell kinetics in soft-tissue sarcomas by PCNA might be a useful adjunct to conventional tumor grading.

Proliferating cell nuclear antigen labeling index of preoperative biopsy specimens in gastric carcinoma with special reference to prognosis.

Proliferating cell nuclear antigen (PCNA) is an auxiliary protein of DNA polymerase delta and is considered to correlate with the cell's proliferative state. Immunohistochemical methods with a anti-PCNA antibody (PC10) have particular advantages compared with other techniques because of the maintenance of the tissue architecture and simplicity of the methodology. The authors investigated the correlation between the tumor's proliferative activity and progression of gastric carcinoma using immunohistochemical staining with PC10. One hundred eighty-nine endoscopic biopsy specimens from patients with gastric carcinoma before operation were investigated by immunohistochemical study, using anti-PCNA monoclonal antibody (PC10). The correlation of PCNA labeling index (percentage of positive cells in more than 500 tumor cells) with various clinicopathologic factors and prognosis was studied. The PCNA labeling index became higher as the histologic stage increased. In patients with lymph node metastasis, vascular invasion, liver metastasis, and peritoneal metastasis, the labeling index was significantly higher. In patients with high PCNA labeling index (50% or greater), prognosis was significantly poorer than in those with a low index (less than 50%). The rate of disease recurrence after curative resection was significantly higher in patients with high PCNA grade group. As a result of this study, PCNA labeling index is suggested to be correlated with the depth of invasion, organ metastasis, vascular invasion, and tumor stage and to be effective as a prognostic factor.

Prognostic significance of PCNA grade in malignant melanoma.

Proliferating cell nuclear antigen (PCNA) grade (a semi-quantitative assessment of the entire tumor profile) and estimation whether PCNA grade is low (0-50% positive cells) or high (51-100% positive cells) has been demonstrated to be of prognostic significance in gastric carcinoma. Sixteen vertical growth phase melanomas with 8 or more years of clinical follow-up were reacted with antibody to PCNA. PCNA grade was independently evaluated by both authors without knowledge of clinical outcome. PCNA grade was subsequently correlated with clinical outcome and other prognostic indicators. Agreement in assigning PCNA grade was noted between the two observers in 13/16 cases. However, PCNA grade failed to accurately predict patient survival or death. Additionally, PCNA grade was not significantly correlated with other recognized prognostic attributes. While interpretation of the prognostic significance of PCNA grade based upon this small series of cases should be conservative, a semi-quantitative estimate of PCNA-positive cells appears to have little utility in predicting the clinical outcome in malignant melanoma.

Prognostic value of proliferating cell nuclear antigen in lymph node—negative breast cancer patients

The prognostic value of proliferating cell nuclear antigen (PCNA) has been demonstrated in recent studies of human tumors including breast cancer. PCNA immunoreactivity was assessed retrospectively in a consecutive series of 173 lymph node-negative primary breast cancer cases. The PCNA grade was determined according to estimated quartiles of nuclear immunostaining, and its association to disease-free and overall survival was studied. PCNA grade was associated significantly with nuclear grade. On univariate analysis, PCNA grade was associated significantly with both 5-year relapse-free survival rate (Grades 1-2 = 78%; Grades 3-4 = 52%; P = 0.0117) and overall survival. On multivariate analysis, T category and PCNA were associated independently and significantly with both relapse-free and overall survival. Nevertheless, the magnitude of the association of PCNA grade to prognosis was low (relative risk of recurrence in patients with PCNA Grades 3-4 versus Grades 1-2 = 2.13), and only 24% of all relapses or 28.8% of all cancer deaths observed at 5 years occurred in patients with PCNA Grades 3-4. The predictive value of the relationship of PCNA grade to prognosis was too low to be of value as an independent prognostic indicator.

Proliferating cell nuclear antigen immunoreactivity in cervical intraepithelial neoplasia and benign cervical epithelium.

In the normal ectocervix, mitoses are rare and are usually confined to the basal layers. In contrast, they occur more frequently in cervical intraepithelial neoplasia (CIN) and are seen at higher levels, suggesting that CIN may be associated with a progressive dysfunction in proliferative activity of cervical cells. The objective of this study was to use proliferating cell nuclear antigen (PCNA) immunohistochemistry to examine the proliferative activity of cervical epithelial cells in CIN lesions. Sixty-eight cervical biopsies were examined; 20 were totally benign, 14 had CIN I, 21 CIN II, and 13 CIN III. In benign epithelia, PCNA staining was usually confined to the basal layers, whereas in CIN the staining was seen at progressively higher levels of the epithelium. There was a statistically significant correlation between the CIN grade and the highest level of PCNA staining (PCNA grade, r = 0.746, P < 0.001). In addition, the PCNA grade showed significant correlation with the highest level at which mitoses were seen (mitosis grade, r = 0.706, P < 0.001), and a strong direct correlation between the mitosis and CIN grades was also observed (r = 0.955, P < 0.001). These data demonstrate that (1) PCNA immunoreactivity in neoplastic cervical epithelium is different from that seen in the normal cervix, suggesting that CIN is associated with a dysfunctional proliferation of cervical epithelium, (2) that there is a significant correlation between the PCNA grade and CIN grades, and (3) the "mitosis grades" have a strong correlation with the CIN grades.

Prognostic significance of proliferating cell nuclear antigen (PCNA) expression in non-small cell lung cancer.

Formalin-fixed, paraffin-embedded tissue sections of resected tumors from 90 patients were immunohistochemically studied to assess the prognostic value of proliferating cell nuclear antigen (PCNA) expression in non-small cell lung cancer. The individual tumors were classified into groups of high, moderate or low proliferative grade, and 38 (42.2%) patients had a high grade of proliferation. No statistically significant correlations were observed between PCNA grade and TNM status, pathological stage, resectability, histological type, degree of histological differentiation. Only vascular invasion significantly correlated with proliferative grade (p < 0.05). Survival analysis showed that patients with low proliferative grade tumors survived significantly longer (a 5-year survival rate of 83.3%) than those with high proliferative grade tumors (39.4%, p < 0.005). Cox's multivariate analysis revealed that PCNA grade was a significant prognostic determinant of survival. These results suggest that PCNA expression provides an independent prognostic variable for patients with non-small cell lung cancer and that it may be useful to consider this factor in treatment planning.

Cell proliferation in breast carcinoma assessed by a PCNA grading system and its relation to other prognostic variables

This study was undertaken to relate the expression of the proliferating cell nuclear antigen (PCNA), a proliferation marker of putative prognostic significance, to some more established prognostic factors in a series of 60 consecutive breast cancer surgical specimens. PCNA was detected by the PC10 monoclonal antibody (MAb) using an immunohistochemical method and PCNA immunostaining was estimated on a semi-quantitative basis, a cut-off value of 50% of positively stained tumour cells discriminating between the high (> 50%) and low (< 50%) PCNA grade. The PCNA grade did not correlate with tumour size and axillary node status. However, a high PCNA grade tended to be associated with a poor histological grade and there was an inverse relationship with oestrogen-receptor status, as determined by means of the immunohistochemical staining for the oestrogen-induced pS2 protein. These conflicting results suggest that the possible prognostic usefulness of PCNA immunostaining, as a measure of cell proliferation rate, in breast cancer is yet to be demonstrated and can be validated only by direct relation to survival data.

Delay in fixation does not affect the immunoreactivity of proliferating cell nuclear antigen (PCNA).

The effect of delayed fixation on the immunoreactivity of proliferating cell nuclear antigen (PCNA) was investigated using eight breast carcinomas. Topologically shuffled samples of each tumour were immersed in fixative at times of 0.5, 1, 2, 4, 6, 18, and 24 h after surgical removal. In addition to a PCNA index (percentage of positive cells per 1200 tumour cells), a semi-quantitative PCNA grading system was used, based on estimates of more than or less than 50 per cent of positive tumour cells at each time interval. The PCNA index of six tumours increased by a mean of 10 per cent with a fixation delay of 24 h. The PCNA grade of all eight tumours showed no change with delayed fixation.

Prognostic value of proliferating cell nuclear antigen in gastric carcinoma.

A new monoclonal antibody to proliferating cell nuclear antigen (PCNA), PC10, which can be used on routinely processed tissue, was applied to 93 cases of gastric carcinoma. Significant intra-tumoural variation in staining occurred. In addition to a PCNA index (percentage of positive cells per 1000 tumour cells), a semiquantitative PCNA grading system was devised, based on estimates of less than or more than 50% of positive tumour cells in whole sections. Neither PCNA index nor PCNA grade showed any correlation with established histological variables, tumour stage, or the presence of lymph node metastases. No significant correlation was observed between PCNA index and S + G2M phase fraction measured by flow cytometric analysis. To analyse survival tumours with PCNA indices above and below the median level (41%) were compared. Those with a higher index tended to have a worse prognosis, but when PCNA grade was considered, it was found to have definite independent prognostic value, tumours of low grade surviving better than those of high grade. The ability of semiquantitative PCNA grading to allow for intra-tumoural variation suggests it may have advantages over absolute counting, which is prone to sampling error when tumour heterogeneity is a major factor. The prognostic value of PC10 staining in gastric carcinoma is therefore promising.