A comparative study on aberrations of chromosome 17 and proliferating cell fraction in lung cancer.

To better understand the relationship between specific chromosome changes found in human lung tumors and their phenotypic consequences a the tissue level, an in situ hybridization (ISH) procedure of chromosome 17 and immunohistochemistry of proliferating cell nuclear antigen (PCNA) were done. The deparaffinized sections were stained with pericentromeric probes for chromosome 17 and an immunohistochemical study of a monoclonal antibody against PCNA were performed. The numbers of chromosome signals were than compared with the positivity of PCNA expression. The mean numbers of chromosome were 1.62 in normal lymphocytes and 2.48 in lung cancer cells. Tumors showed a high mean positivity of PCNA of 43.4%. Mean PCNA expression was higher in squamous carcinomas than in adenocarcinomas (p < 0.05). A linear correlation between numbers of ISH signals and PCNA expression was not demonstrated, but there was a tendency of increasing PCNA positivity according to increasing numbers of ISH signals in adenocarcinomas of the lung and the tumor tissues which were over 50% positive PCNA expression. There was no linear correlation between numbers of ISH signals, PCNA positivity and tumor stages, and keratinization of squamous cell lung cancer. These results suggest that ISH will prove to bo an important tool for determining the underlying genetic basis for tissue phenotypic heterogeneity by allowing genetic determinations to be made on paraffin-embedded tissue sections where histologic architecture is preserved, and immunohistochemical nuclear staining with anti-PCNA on routinely processed tissue is a simple technique for the assessment of proliferation in non-small cell lung carcinoma.     

Proliferating cell nuclear antigen expression in central nervous system neoplasms

Summary Proliferating cell nuclear antigen (PCNA) is a cell-cycle-regulated protein, which can be demonstrated in routinely fixed specimens. Studies on various tissues, cell cultures and neoplasms have shown that PCNA labelling index (LI) correlates with flow cytometry, tritiated thymidine LI, bromodeoxyuridine (BrdU) incorporation and Ki67 LI. PCNA LI may have prognostic value in various neoplasms. The present study concerns PCNA immunostaining in a series of neuroglial tumours. We demonstrate that there is a relation between PCNA LI and histological grade, and between PCNA LI and reported thymidine LI, BrdU LI and Ki67 LI. Pleomorphic xanthoastrocytomas and lowgrade astrocytomas had the lowest LI, whereas metastases of small cell lung cancer and medulloblastomas had the highest LI. Glioblastomas sometimes showed a certain degree of intratumoral heterogeneity of distribution of immunostained cells. Intratumoral heterogeneity underscores the critical importance of representative sampling of central nervous system neoplasms for kinetic studies. As expected, PCNA LI are somewhat higher than tritiated thymidine LI, BrdU LI and Ki67 LI because PCNA is a marker of G1, S, G2 and M-phases of the cell cycle and not of S-phase only. In addition, because of its long half-life, PCNA may be detected immunohistochemically in cells that have recently left the cell cycle. The immunohistochemical evaluation of PCNA LI is easy to perform on routinely processed material, allowing retrospective studies. PCNA LI may be a useful tool in grading gliomas. However, its prognostic value must be validated by comparing PCNA LI with the follow-up of the neoplasms, and possibly with the responsiveness to anti-proliferative therapy.     

p53 expression in oat and non-oat small cell lung carcinomas: correlations with proliferating cell nuclear antigen.

AIMS--To investigate the immunohistochemical expression of p53 protein in small cell lung carcinoma (SCLC), comparing it with that in non-small cell carcinoma (NSCLC); and to evaluate the correlation between p53 and proliferating cell nuclear antigen (PCNA) expression as well as between p53 and PCNA expression and survival. METHODS--Paraffin wax embedded tissues from 61 cases of primary lung carcinoma were stained for p53 protein and PCNA using the monoclonal antibodies 1801 and PC-10, respectively, in a standard avidin-biotin immunoperoxidase method. RESULTS--Of the 61 lung carcinomas 35 (57%) were positive for p53 (range 1% to 90%). Ninety percent of the non-oat SCLC contained positive cells and the labelling index (LI) was significantly higher than that of the oat SCLC (p < 0.001). SLCC also displayed a higher p53 LI than NSCLC (p < 0.01); no difference was found between squamous carcinoma, adenocarcinoma, and oat cell carcinoma. A p53 LI of greater than 1% tended to be associated with nodal metastases (p < 0.05), and p53 expression in node positive tumours as well as in oat cell carcinomas was indicative of poorer survival (p < 0.01 and p < 0.1, respectively). A p53 Li of greater than 60% was a negative prognostic factor in non-oat SCLC (p < 0.05). PCNA LI was highest in non-oat SCLC and lowest in NSCLC; oat cell carcinomas had a mean LI intermediate between NSCLC and non-oat SCLC (NSCLC v oat cell carcinoma p < 0.05 and oat cell v non-oat cell carcinomas p < 0.01). A PCNA LI was not correlated with nodal metastases or survival, but there was a significant positive correlation between PCNA LI and p53 LI (rs = 0.484, p < 0.001). CONCLUSIONS--p53 and PCNA expression differ substantially among the various types of lung carcinomas. Substantial differences were also found between oat and non-oat types of SCLC, indicating that SCLC is heterogeneous as far as proliferation rate and altered p53 expression is concerned. p53 seems to be of some prognostic value. The relation between PCNA and p53 expression indicates that the PCNA gene is slightly upregulated by p53.     

Proliferation indices in spontaneous canine lung cancer: proliferating cell nuclear antigen (PCNA), Ki-67 (MIB1) and mitotic counts

Proliferation indices were measured for specimens from 55 spontaneous canine lung tumours, collected by surgical biopsy from clinical patients and archived in paraffin wax blocks. These indices were then related to the mitotic index and histological type of the tumour. Proliferating cell nuclear antigen (PCNA) and Ki-67 (MIB1) proteins were detected immunohistochemically with a biotin-streptavidin amplified detection system on a representative tissue section from each tumour. Five adjacent, non-overlapping fields were selected at random, and 200 cells per field were examined in each section. For PCNA, cells were classified subjectively into negative, weak or strong reactivity groups, based on nuclear staining. MIB1 cells were classified as negative or positive, based on nuclear staining. Mitotic figures were counted in anti-PCNA-labelled and anti-MIB1-labelled sections, in the same tumour areas as those in which indices were established. Mitotic counts were also done on haematoxylin and eosin-stained tissue sections. Linear regression analysis showed that all PCNA, MIB1 and mitotic indices had highly significant positive correlations (P<0.0001) with each other. Adenosquamous and squamous cell carcinomas differed from other histological tumour types in having significantly higher proliferation indices. These data suggest that growth rates for lung tumours in the dog vary according to histological type. On the basis of differences in proliferation indices and the distribution of immunoreactivity between histological subtypes in this study, it would seem that immunohistochemical detection of PCNA and MIB/1 reactivity and analysis of mitotic figures in routinely processed tissues may be useful in the diagnosis of lung tumours. 1999 W.B. Saunders and Company Ltd.     

Apoptosis occurs independently of bcl-2 and p53 over-expression in non-small cell lung carcinoma

Certain oncogenes and tumour suppressor genes are known to modulate apoptosis. To investigate whether over-expressed bcl-2 and abnormally stabilized p53 are associated with reduced apoptosis in paraffin sections of non-small cell lung carcinoma, apoptotic, mitotic, and Ki-67 labelling indices were determined and correlated with bcl-2 and p53 immunoreactivity in 54 squamous cell carcinomas and 22 adenocarcinomas. Nineteen squamous cell carcinomas (35.2%) showed over-expression of bcl-2, but all 22 adenocarcinomas were bcl-2 negative. Thirty-seven squamous cell carcinomas (68.5%) and 13 adenocarcinomas (59.1%) showed p53 over-expression. Apoptotic tumour cells were identified among p53 positive and bcl-2 positive tumour cells. There was a significant linear correlation between apoptotic indices and mitotic indices. bcl-2 over-expression and p53 over-expression were not associated with attenuated apoptosis, or altered mitotic or Ki-67 labelling indices in either tumour type. Neither bcl-2 nor p53 was of prognostic significance. These results suggest that apoptosis in non-small cell lung carcinoma occurs independently, and is not modulated primarily by, bcl-2 or p53. It is likely that the effects on apoptosis of bcl-2 and p53 are countered by those of other oncogene products and/or additional factors that regulate apoptosis in vivo     

Comparison of bromodeoxyuridine and proliferating cell nuclear antigen labeling in gastric carcinoma.

The proliferative activity of human gastric carcinoma was measured by means of in vitro incorporation of the thymidine analogue, 5-bromo-2''-deoxyuridine (BrdU), into the newly-synthesized DNA of fresh tumors and immunohistochemical staining of proliferating cell nuclear antigen (PCNA) using avidin-biotin peroxidase method. Eighty-two cases of surgically resected human gastric carcinomas consisting of 18 various histologic types were subjected to study. The mean BrdU labelling index (LI) and PCNA LI were 22.9% and 39.1%, respectively. The correlation between BrdU LI and PCNA LI was statistically significant (correlation coefficient mu = 0.61334, p = 0.0001). We concluded that immunohistochemical staining for PCNA may become a practical method instead of in vitro or in vivo BrdU labeling to assess the proliferation fraction of the gastric cancer patient.     

Apoptosis and proliferation in relation to histopathological variables and prognosis in hepatocellular carcinoma

The prognosis of patients with resectable hepatocellular carcinoma depends mainly on the anatomical extent of the tumour and on the general condition of the patient. Given the growing evidence that proliferation indices may be of prognostic significance in hepatocellular carcinomas and that parameters of cell loss (usually, but not exclusively, due to programmed cell death) are biologically relevant, the identification and quantitation of proliferative capacity and apoptosis may be of prognostic importance. In this study four different methods have been used to assess proliferation in a series of 193 curatively (R0) resected hepatocellular carcinomas: mitotic count, immunohistochemical assessment of MIB-1 (Ki-67), proliferating cell nuclear antigen (PCNA), and silver-stained nucleolar organizer regions (AgNORs). Apoptosis was assessed using the in situ-end labelling (ISEL) technique in combination with morphological criteria. Patients who received liver transplantation were excluded. The results obtained were compared with histopathological stage (according to UICC), Edmondson grade, several other histopathological factors, and survival rate. Significant statistical correlations were seen between the mitotic index, the rate of nuclear positivity for MIB-1 and PCNA, and the number of AgNOR dots. In univariate survival analysis, tumour stage and Edmondson grade, mitotic index, MIB-1 and PCNA index, and mean AgNOR number were significant factors influencing patients' survival. On multivariate Cox survival analysis, mitotix index, concomitant cirrhosis, Edmondson grade, and patient age were the only significant independent prognostic factors. Apoptosis was not related to prognosis or to other parameters examined. These results indicated that mitotic index is an additional prognostic parameter which could provide auxiliary information for patients' outcome. MIB-1 and PCNA immunostaining and AgNORs showed a good correlation among themselves. Apoptosis did not predict prognosis in hepatocellular carcinoma. Copyright © 1999 John Wiley & Sons, Ltd.     

Determination of proliferating fractions in malignant melanomas by anti-PCNA/cyclin monoclonal antibody

An immunohistochemical study of melanocytic tumours using 19A2, a monoclonal antibody against proliferating cell nuclear antigen (PCNA/cyclin), was performed on tissues routinely processed with formalin fixation and paraffin embedding. In normal skin, keratinocytes of the suprabasal region in epidermis, the papillae and outer root sheath of hair follicles and the basal cells lining the lobules of sebaceous glands were stained in the nucleus. Other skin components, including basal and follicular melanocytes, did not demonstrate nuclear labelling. In addition, expression of PCNA/cyclin in keratinocytes was higher in sun-exposed skin compared with unexposed skin. In melanocytic lesions, PCNA/cyclin positive tumour cells increased in number and staining intensity according to the following progression: common melanocytic naevi; dysplastic naevi; primary melanomas; and metastatic melanomas. Expression of PCNA/cyclin, therefore, provides a useful marker for proliferation and tumour progression in skin.     

Proliferating cell nuclear antigen and S phase fraction in endometrial stromal sarcoma.

AIMS: To investigate the value of immunohistochemical staining for the cell cycle protein proliferating cell nuclear antigen (PCNA) and flow cytometric S phase fraction in determining prognosis in endometrial stromal sarcoma, graded according to mitotic count. METHODS: Seventeen endometrial stromal sarcomas from 13 patients treated at the Royal Marsden Hospital were analysed. Serial 5 microns sections were cut for haematoxylin and eosin and immunohistochemical staining for PCNA, performed using the murine monoclonal antibody PC10. PCNA positivity was expressed as a percentage of the total number of cells (PCNA index). Flow cytometric analysis was performed on nuclei extracted from paraffin wax sections. RESULTS: In the five patients who died of disease within five years, PCNA index varied between < 1% and 60% (mean 21%) and S phase fraction ranged from 11.3 and 20.1 (mean 13.8). Four patients who were apparently cured showed PCNA indices ranging from < 1% to 5% (mean 1.75%) and S phase fraction ranging from 1.4 to 3.5 (mean 2.3); and three patients alive with disease showed PCNA indices ranging from 1% to 15% (mean 8.6%) and S phase fraction ranging from 1.4 to 3.5 (mean 2.3). One patient who died from indolent local disease after nine years showed a PCNA of 1 or less and an S phase fraction of 0.9. CONCLUSIONS: PCNA staining was variable and therefore not a reliable prognostic indicator, but a high PCNA index was only found in those patients dying of disease within five years. A stronger association was seen between S phase fraction and prognosis; this also correlated well with histological grade determined by mitotic count. In individual borderline cases that are between low and high grade categories, these procedures may be useful.     

Nuclear markers (star volume, mitotic index, AgNOR and Ki-67) of the primary tumor and its metastasis in non-small cell lung carcinomas

Although tumor growth is controlled by growth rate and cell cycle, it is also likely that the proliferative activity of tumor cells can influence the growth rate of the primary cancer and account for their aggressiveness. Variations in growth rate, cell cycle control and proliferative activity could, in part, explain differences in invasive and metastatic properties among non-small cell lung carcinomas (NSLC). The purpose of this report is to: (1) evaluate growth rate by using growth- and cell cycle-regulating markers (mitotic count, nuclear star volume, AgNOR, and Ki-67) as reflections of growth rate and (2) compare the indices of primary NSLC with the indices of their metastasis in a series of patients with advanced disease. Thirty-three patients with non-small cell lung cancer and hematogenous metastases were retrospectively studied by histochemical, immunohistochemical, and morphometrical investigations. Clinical variables were examined for differences in the frequency of histological subtypes, nuclear star volume, mitotic index, AgNOR area, and Ki-67 immunohistochemistry indices of expression in subgroups of patients stratified by primary tumor and hematogenic metastasis. The impact of these factors on overall follow-up was analyzed. In the samples available in this study, consisting of primary-met paired tumors, which are unique and rarely available for studies of lung cancer, we found that nuclear star volume and mitotic index in metastatic tumors were significantly higher than in the primary tumors. Although there was no significant difference between the Ki-67 index of metastatic and primary tumors, the Ki-67 index in metastatic brain tumors was significantly higher than in the corresponding primary tumors. Examination of Kaplan-Meier survival curves demonstrated that patients with metastatic tumors showing AgNOR area higher than 10.82 microm2 and nuclear star volume lower than 559.50 microm3 had approximately the same odds ratio (log rank of 4.16 and 3.25, p = 0.04 and 0.01, respectively) for survival with a median survival time equal to 17 months for both groups. Analysis of the remaining marker correlation had no impact on survival. We conclude that these results offer future possibilities for more complex studies, including the results of additional immunohistochemical analysis using molecular markers that are known to be important in regulating cell proliferation, e.g., cyclin D1, p27, and cyclin E. These would influence clinical decisions or different therapeutic approaches in advanced stage disease of non-small cell lung cancer.     

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.     

非小细胞肺癌中Mina53和PCNA的表达及意义

目的探讨Mina53在非小细胞肺癌(non small cell lung carcinoma,NSCLC)中的作用及Mina53、增殖细胞核抗原(PC-NA)在NSCLC中联合表达的意义。方法采用免疫组化PV-9000两步法、Western blot法检测46例NSCLC和10例正常肺组织中Mina53、PCNA蛋白的表达情况。结果在正常肺组织中Mina53、PCNA表达量均很少,而在NSCLC中均高度表达。Mi-na53与PCNA表达均与组织分化、临床分期、淋巴结转移有明显的相关性,Mina53、PCNA蛋白之间具有明显的正相关性。结论 Mina53、PCNA相互影响、相互作用,在肺癌的发展过程中起促进作用。     

Differential expression of hypoxia inducible factor-1 alpha and tumor cell proliferation between squamous cell carcinomas and adenocarcinomas among operable non-small cell lung carcinomas

This study aimed to evaluate whether the elevated level of hypoxia-inducible factor-1alpha (HIF-1alpha) correlated with histologic types, angiogenesis, tumor cell proliferation, and clinical parameters in common non-small cell lung carcinomas (NSCLCs). We performed immunohistochemical stains using paraffin-embedded tissue blocks from 84 cases of operable NSCLC [No. of squamous cell carcinoma (SCC), 45; No. of adenocarcinoma (AC), 39]. HIF-1alpha expression was related with histologic types (66.7% in SCCs vs 20.5% in ACs, p<0.001), but not with lymph node status, tumor stage, vascular endothelial growth factor expression, microvessel density (MVD), and proliferating cell nuclear antigen (PCNA) index (p>0.05, respectively). As for the histologic types, MVD and PCNA index were significantly higher in SCCs than in ACs (p=0.009 and p=0.016, respectively). Among HIF-1alpha positive carcinomas, MVD was significantly higher in HIF-1alpha positive SCCs than in HIF-1alpha positive ACs (p=0.023). The overall survival curves were not associated with HIF-1alpha expression or any other histologic parameters (p>0.05). These findings suggest that HIF-1alpha expression in NSCLCs may play a differential role according to histologic types, but its prognostic significance is indeterminate.     

DEVELOPMENT OF AN OPTIMAL PROTOCOL FOR ANTIGEN RETRIEVAL: A ‘TEST BATTERY’ APPROACH EXEMPLIFIED WITH REFERENCE TO THE STAINING OF RETINOBLASTOMA PROTEIN (pRB) IN FORMALIN-FIXED PARAFFIN SECTIONS

The retinoblastoma (RB) gene, which encodes the nuclear RB protein (pRB), is believed to be involved in cell cycle control and cell differentiation. Studies have demonstrated that loss of RB function may play a role in tumour formation and progression of a variety of human tumours, such as bladder, lung, breast, and prostate cancers. The immunohistochemical detection of pRB expression in formalin–paraffin sections of human cancer has potential advantages of convenience, economy, and compatibility with routine surgical pathology practice. In practice, however, results using pRB antibodies on routinely processed, paraffin-embedded tissue have been inconsistent. In this study, the antigen retrieval (AR) method has been applied to the immunohistochemical detection of pRB in paraffin-embedded tissues and a ‘test battery’ approach has been developed to identify the principal variables that result in the optimal AR protocol. This approach includes the use of buffered solutions at pH 1, 6, and 10 with three different heating conditions (temperatures 120°C, 100°C, and 90°C). In the example described here with antibody RB-WL-1, the low pH solution with the microwave heating at 100°C proved most effective. Both fresh and routinely processed formalin–paraffin tissues of normal and bladder carcinoma were used for a comparison of the pRB immunostaining. The AR method was evaluated by comparing the immunohistochemical staining result on routinely processed formalin–paraffin sections with frozen sections of the same tumour. A consistent intensity of immunohistochemical staining for pRB was achieved using the identified optimal AR protocol on formalin–paraffin sections. All slides showed positive staining of pRB in normal mesenchymal and epithelial tissues. The pattern of pRB localization and intensity of staining was similar to that obtained in frozen sections, though the intensity obtained by AR treatment on paraffin sections was slightly to moderately stronger than that obtained in frozen sections. Once the protocol was identified, it was tested using routinely processed paraffin tissue sections of 245 cases of bladder carcinoma, with consistent pRB immunostaining results. The protocol described is simple to perform and gives reproducible results for evaluation of pRB expression by immunohistochemistry.     

Extent of apoptosis in ovarian serous carcinoma: relation to mitotic and proliferative indices, p53 expression, and survival.

AIMS: To assess the extent of apoptosis in ovarian serous carcinoma and to examine possible relations between apoptosis, cell proliferation, p53 overexpression, and patient survival. METHODS: Apoptotic and mitotic indices were obtained by examining haematoxylin and eosin stained sections from 30 patients with ovarian serous carcinoma. Apoptosis was also evaluated semiquantitatively by in situ end labelling of fragmented DNA. Expression of p53 and determination of Ki-67 labelling indices were based on immunohistochemical staining. Clinical details were obtained from patients' clinical records. For statistical analysis, Fisher's exact test, parametric (Pearson) linear correlations, and the Kaplan-Meier method were used. RESULTS: The mean apoptotic index was 1.3% (range 0.02-3.9%), the mean mitotic index was 0.4% (range 0.02-1.1%), and the mean Ki-67 labelling index was 16% (range 4-32%). There were significant correlations between the apoptotic and mitotic indices (p < 0.0205) and between the mitotic and Ki-67 labelling indices (p < 0.024). There was a significant correlation between a high apoptotic index and poor prognosis (p < 0.02). p53 was overexpressed in 16 cases but the extent of apoptosis and outcome were both independent of p53 status. CONCLUSIONS: These results suggest that regulation of apoptosis is an integral component of tumour cell kinetics in ovarian serous carcinoma, and that increased apoptosis is indicative of aggressive tumour growth. p53 expression did not correlate with altered apoptosis, but the possibility of an attenuated apoptotic response to subsequent DNA damage by anticancer agents is not excluded.     

Usefulness of MIB1 monoclonal antibody in assessing the proliferative index in human bladder carcinoma: comparison with Ki-67 antibody

The reactivity of MIB1 antibody on routinely processed paraffin sections was compared with that of Ki-67 antibody on frozen sections of 80 transitional cell carcinomas of the bladder. The percentage of labelled cells was expressed as the labelling index. MIB1 labelling indices were higher than those of Ki-67 but for each case the two values were strongly correlated ( r = 0.91). Ki-67 and MIB1 indices were also correlated to tumour grade and stage (P 0.001). MIB1 indices determined after both formaldehyde and ethanol based Bouin's fluid fixatives did not show any significant difference. MIB1 antibody staining after microwave oven heating of tissue sections is a simple technique for assessing the proliferative fraction of bladder tumours on fixed material. The use of MIB1 antibody permits retrospective studies and should determine whether the proliferation index in bladder carcinoma has the same prognostic value as demonstrated in other types of neoplasms.     

Analysis of cell cycle regulator proteins in non-small cell lung cancer

BACKGROUND/AIMS: Abnormalities of the proteins involved in cell cycle checkpoints are extremely common among almost all neoplasms. This study aimed to investigate the expression of four components of the cell cycle machinery-p21, p16, p53, and proliferating cell nuclear antigen (PCNA)-in non-small cell lung cancer (NSCLC). METHODS: The expression of p21, p16, p53, and PCNA was examined in 68 well characterised NSCLC specimens using immunohistochemistry. The coregulation of these proteins and their influence on survival were analysed using both univariate and multivariate analyses. RESULTS: By univariate analysis, the expression of all the proteins examined, except for PCNA, was significantly correlated with survival. In multivariate analysis, the only immunohistochemical parameter able to influence overall survival was p16, confirming the hypothesis that the RB-p16 tumour suppressor pathway is inactivated in most lung cancer samples. Finally, the group of patients with NSCLC who were negative for both p21 and p16 had a significantly shorter overall survival. CONCLUSIONS: These results suggest that loss of control of cell cycle checkpoints is a common occurrence in lung cancers, and support the idea that functional cooperation between different cell cycle inhibitor proteins constitutes another level of regulation in cell growth control and tumour suppression.     

Peripheral airway cell marker expression in non-small cell lung carcinoma. Association with distinct clinicopathologic features.

Paraffin sections of 247 primary and metastatic non-small cell lung carcinomas, the corresponding non-neoplastic lungs, and 75 other specimens were examined by immunohistochemical procedures using a panel of antibodies against the specific products of peripheral airway cells: the major surfactant-associated protein and 10-kD Clara cell protein. Non-small cell lung carcinoma tumors most frequently positive for either peripheral airway cell marker were adenocarcinomas (41%), especially those with papillolepidic growth pattern (56%), followed by large cell carcinomas (25%), other adenocarcinomas (22%), and squamous cell carcinomas (16%). Immunoreactivity was mainly focal and the expression of the two peripheral airway cell markers was discordant. The incidence of marker expression was similar in metastatic and primary non-small cell lung carcinoma. Other organs and their tumors were negative, with few exceptions. Non-small cell lung carcinomas positive for peripheral airway cell markers were associated with younger age and less-intense smoking, and surfactant-associated protein reactivity was more common in women than in men. Peripheral airway cell markers were independent prognostic factors for survival and delayed development of metastases in patients with less-advanced disease. It is concluded that surfactant-associated protein and 10-kD Clara cell protein are specific markers for non-small cell lung carcinoma and peripheral airway cell differentiation and provide useful tools to study the pathogenesis, biology, and prognosis of non-small cell lung carcinoma.     

Immunohistochemical localization of cathepsin D, proliferating cell nuclear antigen and epidermal growth factor receptor in human breast carcinoma analysed by computer image analyser: correlation with histological grade and metastatic behaviour

 

Aims:

The purpose of this study is to examine the relationship between immunohistochemical localization of cathepsin D (CD), proliferating cell nuclear antigen (PCNA) and epidermal growth factor receptor (EGF-R) in 65 cases of breast carcinoma in Japanese women and traditional prognostic factors such as histological grade, lymph node status, mitotic rate and clinical stage, in order to possibly identify some indicator(s) that may be specifically associated with prognosis.  

Methods and results:

Serial sections of 5-μm thick were cut from the archival formalin-fixed, paraffin-embedded tissue blocks, and processed for CD, PCNA and EGF-R immunostaining. The results were analysed by computer-based image analysis system. All samples showed a positive immunoreaction for cathepsin D in both the parenchyma and stroma. However, the staining area and intensity varied from cell to cell in the parenchyma and stroma as well as among samples. Subsequently, the evaluation of immunostaining for CD was separately performed in both the parenchyma and stroma (CDpar and CDstr, respectively) and the combination of both components (CDtotal). PCNA and EGF-R showed positive immunostaining almost exclusively in the parenchymal component of the carcinoma tissue specimens. CDtotal significantly correlated with the histological grade, PCNA index (PI), mitotic rate (MR), EGF-R and lymph node metastasis. Significant correlation was also demonstrated between CDpar and the histological grade, EGF-R and lymph node metastasis, or between CDstr and MR, EGF-R and lymph node metastasis. EGF-R correlated highly with the histological grade, MR score, lymph node metastases and recurrence-free survival.  

Conclusions:

Both the CD parameters and EGF-R are valuable indicators for predicting the biological behaviour of human breast carcinoma.     

Renal cell carcinoma with rhabdoid features: an aggressive neoplasm

AIMS: Only a few reports on renal cell carcinoma with rhabdoid features have been published. This study was performed to investigate the clinicopathological characteristics of renal cell carcinomas with rhabdoid features. METHODS AND RESULTS: Among 253 cases of renal cell carcinoma in adults, eight cases with rhabdoid features were detected. Rhabdoid areas ranged from 10% to 90% of each of the cases. Seven of the eight cases were TNM stage III or IV, and four of the eight cases died within 8 months of surgery. Immunohistochemically, the rhabdoid areas were positive for CAM 5.2 (4/8), AE1/AE3 (6/8), epithelial membrane antigen (6/8) and vimentin (8/8), and negative for myogenetic markers (0/8). The mean MIB-1 labelling index in the rhabdoid areas was higher than that in the definite carcinomatous areas. Ultrastructurally, perinuclear whorls of intermediate filaments were demonstrated in three of the eight cases using paraffin-embedded blocks. CONCLUSIONS: The rhabdoid areas in renal cell carcinoma have histological, immunohistochemical and ultrastructural similarities to malignant rhabdoid tumours. Renal cell carcinoma with rhabdoid features is a highly aggressive neoplasm and its malignant behaviour may be due to the high cell-proliferative activity of the rhabdoid areas. Rhabdoid features in renal cell carcinoma may represent the endpoint of clonal evolution of renal cell carcinoma (especially in clear cell type cases).