Immunohistochemical diagnosis of prostatic cancer with metastasis

An immunohistochemical method for detecting prostatic acid phosphatase is described for the diagnosis of metastatic prostatic carcinoma. The specific antiserum against prostatic acid phosphatase was prepared from rabbit by injection of acid phosphatase purified from seminal fluid. This method gives a selective staining of the cytoplasm of the glandular epithelial cells of prostatic tissue specimens on paraffin section. Most of the non-prostatic tissues were negative except for occasional weak staining in granulocytes, islet cells of pancreas, parietal cells of stomach, tubular epithelial cells of kidney, and liver cells. Also examined were 50 consecutive cases of metastatic tumor involving the bone marrow and 5 cases of metastatic prostatic carcinoma involving the lymph node or lung. All 20 cases with prostatic primary lesion showed positive staining. All other cases were negative, except 5 of the 14 cases of metastatic breast carcinoma in women showing weakly positive results. The method is fairly specific for identification of metastatic prostatic carcinoma. Occasional positive staining in breast tumor needs further study to establish whether the staining is due to the same isoenzyme or to certain cross immunoreactivity.     

Fucosyltransferase activity in metastasizing and nonmetastasizing rat mammary carcinomas.

Fucosyltransferase levels in 6 established strains of spontaneously metastasizing rat mammary tumors (STMT-058, MT-449, DMBA-4, SMT-077, TMT-081, and SMT-2A) were compared with 4 nonmetastasizing strains (MT-W9B, MT-W9A, MT-100, and MT-66) as controls. Two acceptors were prepared from fetuin for the assay, one by acid hydrolysis of N-acetylneuraminic acid and the other by the stepwise removal of N-acetylneuraminic acid and penultimate galactose by Smith degradation. The enzyme that transfers fucose to the first acceptor was designated fucosyltransferase A, whereas the one that uses the second acceptor was designated fucosyltransferase B. Both types of fucosyltransferases were found in this rat mammary tumor system. Whereas the levels of fucosyltransferase A in the 2 tumor groups were comparable, those of fucosyltransferase B were sixfold to sevenfold higher in the metastasizing tumors. This difference in the level of fucosyltransferase B was not caused either by differential hydrolysis of GDP-fucose by pyrophosphatase in the 2 groups or by hydrolysis of the product by fucosidases. Presence of any other inhibitor(s) or activator(s) of fucosyltransferase was excluded by mixing experiments. Optimal conditions for the assay of this enzyme were determined in a representative strain from each group. Under all circumstances, the activity of fucosyltransferase B was higher in the metastasizing tumors. The enzyme was inhibited by nucleoside diphosphates and triphosphates, and guanosine nucleotides were the most efficient inhibitors. Subcellular distributions of the two fucosyltransferases were similar, 35-50% of the enzyme activity being present in the crude microsomes. When plasma membrane factions were prepared from the microsomes, the major part (50-70%) of the enzyme was associated with the light and heavy plasma membrane fractions. Increased activity of fucosyltransferase B in the group of metastasizing tumors may have reflected faster synthesis and shedding of fucose-containing glycoprotein antigens. Similar molecules possibly were also synthesized in the nonmetastasizing cells but at a much slower rate, because the antigen is not easily lost from the cell surface. Any alteration of the specificity of this focosyltransferase in the metastasizing tumors, in addition, may have caused antigen modulation.     

The induction of NB rat prostatic carcinomas

The NB rat prostate adenocarcinoma model is one in which microscopic carcinoma can be induced via the use of testosterone and estrogen silastic implants. There is an incidence of large, grossly-positive tumors; which approaches 10% to 15%. Seventy-three percent of the animals subjected to this induction method for periods of 9 to 18 months developed at least microscopic carcinoma. The earliest appearance microscopic foci of carcinoma was after three months of implantation, in which one of 30 animals so treated did show carcinoma. However, 88% of the animals treated for 18 months or longer showed microscopic carcinoma.     

Immunohistochemical demonstration of peptide hormones in endometrial carcinomas

Sixty-eight endometrial carcinomas were examined histochemically and immunohistochemically for the presence of amine-containing or neurohormonal peptide-containing cells, particularly in relation to argyrophil cells. Argyrophil cells, detected in 43 of the 68 endometrial carcinomas by the Grimelius method, were subgrouped into two types according to the distribution of argyrophil granules and the shape of the tumor cells. Type I was found in 7 tumors and type II in 39; 3 tumors contained both cell types. The argyrophilia of type II cells was diminished in varying degrees in some tumors by diastase digestion, although it was unchanged in type I argyrophil cells. Indoleamine was detected by the formaldehyde-induced fluorescence method in type I argyrophil cells of four carcinomas. Immunohistochemically, somatostatin-reactive cells were found in two well-differentiated adenocarcinomas with argyrophilia; many of these cells corresponded to some of the type I argyrophil cells, although some were non-argyrophilic. Two adenosquamous cell carcinomas with type II argyrophil cells also contained cells that were immunoreactive with antisera against gastrin; however, they were non-argyrophilic.     

Studies on clonal heterogeneity in two spontaneously metastasizing mammary carcinomas of recent origin

We have studied the clonal heterogeneity of 2 spontaneously metastasizing mammary carcinomas which recently arose spontaneously in C3H/He female retired breeders. Cells of early (2nd to 5th) transplant generations of these tumors were cloned by a combination of semi-solid agarose colony formation and limiting dilution techniques. Growth characteristics of the various clones in vitro and their tumorigenicity in vivo were evaluated. Subsequently, the role of host immunity and of interclonal interactions in regulating growth of the different clones in vivo were analyzed. We found that, whereas all 16 clones isolated from one tumor (T-58) grew rapidly in vivo and in vitro, 10 clones isolated from the second tumor (MT-2) showed a wide disparity in their growth rates in vivo. Taken together, these clones could generally be divided into 3 categories: (1) rapidly growing lines which grew in vivo at rates similar to or higher than those of the parental line; (2) slow-growing lines which grew more slowly than the parental line; and (3) non-growers which failed to produce tumors in vivo with doses of up to 5 X 10(6) cells injected either s.c. or i.v. but grew in vitro at rates comparable to the parental line. No correlation could be established between the various growth potentials exhibited by these tumor lines and tumor cell morphology in vitro and in vivo, as determined by light and electron microscopy. Sublethal irradiation (550-650 R) of young animals prior to tumor inoculation, or before inoculation of tumor cells into old, low NK syngeneic mice, failed to modify the growth of slow-or non-growing lines in vivo, indicating that host cellular defense mechanisms against the clones, if existent, were not mediated by NK or radiosensitive B or T cells. When clonal interactions were studied by the simultaneous injection of different clones in vivo at different s.c. sites, we found that a slow-growing line failed to modify the growth rate of a rapidly growing line, but accelerated the growth of a second slow-growing line injected simultaneously on the contralateral side, and that this enhancement of tumor growth was radioresistant. A mixture of these 2 lines also grew more rapidly than the individual lines alone. Our findings suggest that phenotypic variations in tumorigenicity can be found in clonal lines derived from spontaneous primary tumors and that these variations are not related to cell cycle properties as measured in vitro.(ABSTRACT TRUNCATED AT 400 WORDS)     

Immunohistochemical labelling for prostate-specific antigen in breast carcinomas

Immunohistochemical detection of prostate-specific antigen (PSA) is an aid in determining the prostatic origin of metastatic cells. However, small amounts of PSA have also been found in non-prostatic tissues and tumors, for example in some breast carcinomas, by highly sensitive immunofluorometric methods, but also by immunohistochemistry. Our aim was to evaluate the prevalence and prognostic value of histologically confirmed PSA immunoreactivity in breast carcinoma. Sections of formalin-fixed, paraffin-embedded samples from 171 breast carcinomas were immunostained for PSA. The staining results were compared with the mitotic activity, tumor size, histological grade, steroid receptors and follow-up data. For analysis the material was divided into subgroups according to the patients' age (pre- and postmenopausal). PSA was found by immunohistochemistry in 54 (32%) breast carcinomas. In survival analysis of the whole patient material PSA positivity did not show prognostic value. Among premenopausal patients concomitant estrogen receptor and PSA-negativity proved to be associated with high risk of breast cancer death (RR 6.2), also after adjustment for tumor size, histological grade, and axillary lymph node status. Among postmenopausal patients PSA positivity was associated with progesterone receptor positivity and high differentiation but not with age, nodal status, or mitotic activity. PSA can be detected by immunohistochemistry in a considerable number of breast carcinomas. PSA immunoreactivity alone does not seem to have any value as general prognosticator of breast carcinoma patients. However, concomitant absence of PSA and estrogen receptors was an indicator of unfavourable prognosis among premenopausal patients.     

Pathogenesis and characteristics of spontaneously metastasizing mammary carcinomas and the general principle of metastasis

Chemical mammary carcinogenesis in immunologically attennuated W/Fu female rats, subjected to specific and nonspecific immunostimulations, yielded many spontaneously metastasizing adenocarcinomas with varying degrees of glandular differentiation, and growth rates. The pathogenesis of these tumors suggested that while carcinogens transform target cells, the host immune system endows them with metastatic potential. The metastatic pathways were recognizable as hematogenous, lymphogenous, or hematogenous-lymphogenous combined, according to the capacity of the tumor cells to intravasate the lymphatic and/or vascular channel and traverse the pulmonary artery. The same metastatic pattern can be reproduced with cells from any of the organs involved, indicating that it is inherent in all cells within a given tumor, rather than being determined by the organs they colonize. The biological, biochemical and immunological characteristics of these tumors resemble human breast cancer thus becoming an effective tool in the formulation of the general principle of metastasis by malignant solid tumors.     

Surface membrane O-alkyl lipid concentration and metastasizing behavior in transplantable rat mammary carcinomas

Whole cell and surface membrane O-alkyl and alk-1-enyl lipid concentrations of 11 transplantable rat mammary carcinomas were measured. A correlation was found between surface membrane O-alkyl lipid levels and metastasizing behavior. No relationship was found between alk-1-enyl lipid concentrations and metastasizing behavior. A possible correlation was found between the product of growth rate and rank order of metastasizing behavior versus surface membrane O-alkyl lipid concentration. Growth rate alone did not correlate with surface membrane O-alkyl lipid concentration.     

Biochemical properties of cyclic nucleotide phosphodiesterase in metastasizing and nonmetastasizing rat mammary carcinomas.

The biochemical properties of cyclic nucleotide phosphodiesterases in a nonmetastasizing and a spontaneously metastasizing rat mammary carcinoma were compared. The phosphooiesterases in both tumors had a pH optimum of around 8.0 and preferentially hydrolysed cyclic purine nucleotides. The rate of hydrolysis of purine nucleotides in the nonmetastasizing tumor was two times higher than in the metastasizing tumor, but the rate of pyrimidine nucleotide hydrolysis was equal in both tumors. Theophylline, caffeine, and D,L-4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (Ro20-1724) inhibited the enzyme activity in both tumors; the percent inhibition was the same by each inhibitor. The cyclic nucleotie phosphodiesterase activity in either tumor was stimulated by Mg++, Mn++, and Co++ and suppressed by Ca++, Zn,++, and Ni++. EDTA inhibited the activity below the basal level (activity in the absence of added cation), an this inhibition could be recovered up to the basal level by an equimolar quantity of either Mn++ or Mg++. Further stimulation of the enzyme activity with increasing concentrations of divalent cations was observed only with Mn++. Similar effects were observe with ethylene glycol bis(beta-aminoethyl ether)-tn,n-tetraacetic acid. The stimulatory cations affected both the low and high Michaelis constant (tkm) enzymes in these tumors by increasing the maximum velocity. In the low Km enzyme, the Km was also slightly increased. Neither guanosine 3',5'-cyclic monophosphate nor adenosine 3',5'-cyclic monophosphate had any effect on the hydrolysis of the other at physiologic levels.     

Immunohistochemical profile and treatment of uncommon types of thyroid carcinomas

The distinction of uncommon types of thyroid carcinomas is important, because their treatment and prognosis differ. The aim of this study was to describe retrospectively the immunohistochemical profile of uncommon types of thyroid carcinomas and mode of treatment. Of the 1194 patients with thyroid carcinomas treated in Rabin Medical Center from 1954 to 2001, 153 were uncommon types (not papillary or follicular carcinomas). Specimens from archival tissue obtained from thyroidectomies in all these cases were revised and immunohistochemically examined. Anaplastic carcinomas (n=59) were positive for high molecular weight cytokeratin (CK HMW), low molecular molecular weight cytokeratin (CK LMW), cytokeratin (CK) 7, CK 8 and 18, thymoglobulin, EMA and vimentin; medullary carcinomas (n=39) were positive for CK LMW, CK 19, CK 8 and 18, CK 10, CK 7, carcinoembryonic antigen (CEA) and calcitonin; Hurthle cell carcinomas (n=30) for CK LMW, CK 19, CK 8 and 18, thyroglobulin, epithelial membrane antigen (EMA) and CEA; squamous cell carcinomas (SCC) (n=12) for CK HMW and cytokeratin total (CKs); lymphomas (n=7) for leukocyte common antigen (LCA) and B-cells (CD 20), and clear cell carcinomas (n=6) for CK LMW, CEA and thyroglobulin. Use of an immunohistochemical panel has diagnostic value in the differentiation of uncommon types of thyroid carcinoma, which help to plan the best mode of treatment.     

Imaging diagnosis of hepatocellular carcinomas

With the rapid progress of various imaging methods, including ultrasonography (US), computed tomography. (CT), digital subtraction angiography (DSA) and magnetic resonance imaging (MRI), it has become possible to detect small liver cancer less than 2 cm in diameter, and the prognosis of hepatocellular carcinomas is now improving rapidly. However, the accurate detection of smaller lesions about 1 cm in diameter and their differential diagnosis are difficult by conventional imaging methods such as US, CT and arteriography. For this purpose, we stressed the effectiveness of the combined use of CT and arteriography (the so-called CT arteriography), CT during arterial portography or Lipiodol CT. The promising future of MRI in this field is also discussed.     

Immunohistochemical study of thyroglobulin in thyroid carcinomas with monoclonal antibodies

The effectiveness of an immunoperoxidase technique using four monoclonal antibodies (mAb) is compared to a technique using one polyclonal antibody (pAb) to detect human thyroglobulin (Tg) in paraffin sections of 55 thyroid carcinomas. With the pAb, a positive reaction was found in 82% of the cases. With the four mAb, the presence of Tg was demonstrated in 96.5% of the cases. The mAb gave better results than the pAb on poorly differentiated and anaplastic thyroid carcinomas. Many of the thyroid carcinomas in this study, especially the poorly differentiated and anaplastic type, failed to react with all four mAb to Tg. These results confirm the notion of the heterogeneity of Tg in thyroid carcinomas and indicate that a battery of carefully selected mAb can be successfully used for routine histopathologic detection of Tg in these tumors.     

Cytological diagnosis of prostatic cancer]

The authors has made 836 cytological assays in 184 patients with adenoma and cancer of the prostate. Eighty patients were examined using the complex technic (the puncture material and secretion of the prostate, and urine for the presence of atypical cells). Cytographs characteristic of such prestatic lesions are described.     

Immunohistochemical studies of blood group-related antigens in human superficial esophageal carcinomas

A total of 63 surgically resected esophageal carcinomas (including 49 superficial esophageal carcinomas) and histologically normal tissue adjacent to the superficial carcinoma (nontumorous epithelium) were examined immunohistochemically for the blood group antigens (BGA) A, B, H, Lewisa, Lewisb, Lewisx, and Lewisy. Deletion of an expected A, B or H antigen occurred in 12 (24.5%) of the 49 superficial carcinomas and three (21.4%) of the 14 advanced carcinomas. Incompatible expression of an unexpected A or B antigens occurred in only one case (1.6%) in the carcinoma. In the clinicopathologic study, there was a significant correlation between immunoreactivity of Lewisa and depth of cancer invasion (chi-square test, P less than 0.05). In the superficial carcinoma, there were significant correlations between immunoreactivity of Lewisx and lymph node status (chi-square test, P less than 0.05), immunoreactivity of Lewisy and prognosis (Z test, P less than 0.05), and incompatible expression of Lewisb for tumor against nontumorous epithelium and histologic variation (chi-square test, P less than 0.01). The functional significance of alternations in BGA expression that may be associated with oncogenesis is not clear. However, immunohistochemical determination of BGA may be a more advantageous marker to predict the patient's clinical course in superficial esophageal carcinoma.     

Immunohistochemical expression of EGFR and p-EGFR in oral squamous cell carcinomas

Epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor of the ErbB family, which is expressed or highly expressed in a variety of solid tumors, including oral cancers. High EGFR expression has been correlated with tumor size, metastasis and survival. In recent years, EGFR has been considered a promising target for monoclonal antibody therapy. A total of 52 patients with oral squamous cell carcinoma (OSCC) were selected for EGFR and phosphorylated EGFR (p-EGFR) detection. Immunohistochemical staining was performed to evaluate EGFR and p-EGFR expression. Positive EGFR and p-EGFR staining was present in 92.3% (48/52) and 98.0% (51/52) of all cases, respectively. High EGFR and p-EGFR expression was present in 63.4% (33/52) and 69.2% (36/52) of all cases, respectively. EGFR and p-EGFR expression did not correlate with the clinical factors tumor stage, regional lymph node metastasis, or distant metastasis. However, a statistically significant correlation was identified between high EGFR expression and the pathologic factor tumor invasion. As a conclusion, the majority of OSCCs highly express EGFR and p-EGFR, indicating the importance of studying the efficacy of anticancer therapy targeting these signal factors.(Pathology Oncology Research Vol 12, No 2, 87–91)     

Immunohistochemical diagnosis in fine-needle aspiration cytology

This paper reports 25 kinds of polyclonal or monoclonal antibodies by ABC immunohistochemical technique used for 253 cell smears by fine-needle aspiration. The results were,1. Immunohistochemical diagnosis were classified into 136 metastatic cancers ( K12 EMA CEA LCA-),92 lymphomas (LCA k12- EMA- CEA-), 4 mesenchymal tumors (Vimentin ), 3 melanomas (S-100 NSE ). 15 reactive proliferations (k λ4 CD CD8 ) and 3 unspecified.2. The origin of 70 metastatic cancers were classified into 36 lung (HLC3-AB ), 4 gastrointestinal tract (MG7 ), 8 thyroid (TGB ), 1 prostate (PSA ), 3 liver (AFP ) and 14 unknown. 3. Immunologic phenotype of 87 lymphomas wereclassified into 66 cases of B-cell, 4 T-cell, 3 hsitocyte, 7 Hodgkin' s diseases and 7 unclear. The above results suggest that immunohistochemlcal method may be used as a new method of diagnosing and differentiating epithelial and non-epithelial tumors, detecting primary focus of metastatic cncer, differentiating between reactive proliferation adn lymphome a     

Overexpression of telomerase-associated chaperone proteins in prostatic intraepithelial neoplasia and carcinomas

Over 90% of prostate cancers express telomerase activity. In an experimental model, hsp90 and p23, which are necessary for telomerase assembly and function, dramatically increase during tumorigenic conversion. We immunohistochemically analyzed 60 prostate carcinomas, 50 prostatic intraepithelial neoplasias (PIN) and 25 benign prostatic tissues to determine whether hsp90/p23 expression correlates with advancing stage and whether chaperone distribution overlaps with hTERT, the catalytic component of telomerase. Strong expression of hsp90/p23 was detected in approximately 95% of PIN and carcinomas without relationship to Gleason score. While hsp90/p23 immunostaining was predominantly diffuse and cytoplasmic, nuclear immunoreactivity was observed in several moderate-to-high grade carcinomas, and those carcinomas with nuclear chaperone staining exhibited detectable hTERT. Our data suggest enhanced chaperone-mediated telomerase assembly as a mechanism for increased activity in advanced prostate carcinomas, stable association between chaperones and telomerase in vivo, and utility for chaperone immunostaining to identify focal PIN in the context of widespread hyperplasia.