Immunohistochemical profile for unknown primary adenocarcinoma

Background

Development of tailored treatment based on immunohistochemical profiles (IPs) of tumors for cancers of unknown primary is needed.

Methodology/Principal Findings

We developed an algorithm based on primary known adenocarcinoma for testing sensitivity and specificity. Formalin-fixed paraffin-embedded tissue samples from 71 patients of unfavorable subsets of unknown primary adenocarcinoma were obtained. We examined 15 molecular markers using the algorithm incorporating these IPs and classified the tumours into 9 subsets based on the primary tumour site. The sensitivity and specificity of this algorithm were 80.3% and 97.6%, respectively. Apparent primary sites were lung in 17 patients, digestive organs in 13, gynecological organs in 9, prostate in 7, liver or kidney in 6, breast in 4, urothelial organ in 2, biliary tract and pancreatic profile in none, and unclassified in 13. The response rate to chemotherapy was highest for the gynecological IPs. Patients with gynecological or lung cancer IPs had longer median progression-free survival than those with others: 11.2 months for gynecological IPs (p<0.001) and 6.8 months for lung IPs (p = 0.05). Lung, digestive, prostate, and gynecological profiles were associated with significantly longer median survival time than the other profiles. Multivariate analysis confirmed that the IPs were independent prognostic factors for survival.

Conclusions/Significance

The IPs identified in this study can be used to further stratify patient prognosis for unfavorable subsets of unknown primary adenocarcinoma.     

Immunohistochemical differentiation of malignant mesothelioma, mesothelial hyperplasia and metastatic adenocarcinoma in serous effusions, utilizing staining for carcinoembryonic antigen, keratin and vimentin

The cytologic diagnosis of malignant mesothelioma and its distinction from mesothelial hyperplasia and metastatic adenocarcinoma is consistently difficult; tissue studies utilizing the immunohistochemical profiles of carcinoembryonic antigen (CEA) and keratin have demonstrated a reproducible distinction between these tumors. Mesothelium contains vimentin in addition to keratin, but its characterization is hindered by its poor preservation in formalin fixatives; alcohol fixation is far superior. Alcohol-fixed, Papanicolaou-stained smears of serous fluids from five cases of reactive mesothelium, five metastatic adenocarcinomas and five malignant mesotheliomas were stained with polyclonal CEA, antikeratin monoclonals AE1 and AE3 (combined) and monoclonal vimentin utilizing the peroxidase-antiperoxidase method. The study revealed the excellent preservation of mesothelial vimentin staining in all three groups. The reactive mesothelium and mesothelioma groups were strongly positive for vimentin and keratin whereas the metastatic adenocarcinoma group was only positive for keratin and CEA (except one case). These findings support the results of previous tissue studies, disclosing CEA staining in the metastatic adenocarcinomas, but not in the mesotheliomas, and the inability of keratin staining to distinguish between the two. The findings also emphasize that positive vimentin staining will usually exclude a metastatic adenocarcinoma, but will not distinguish between neoplastic and reactive mesothelial states.     

Inverse relationship between Leydig cell density and metastatic potential of prostatic adenocarcinoma.

PURPOSE: Evaluate the relationship between metastatic potential of prostatic adenocarcinoma (PC) and testicular Leydig cell density. MATERIALS AND METHODS: Tissue samples from 111 men, age 52-85, with PC and bilateral orchiectomy were evaluated for Leydig cell density. The patients were divided into two groups: Group A were patients with metastasis (n = 36) and Group B were patients without metastasis (n = 75). Leydig cell density was determined by direct manual microscopic cell count on the tissue sections. The means of cell counts by four pathologists, expressed as cell/0.78 mm2 were used for analysis. The normally distributed data were analyzed by two-tail Student's t-test. Thirty-eight age-compatible autopsy cases who died of unrelated causes served as normal controls. RESULTS: The mean of Leydig cell count in group A patients was 14.43 (14.43+/-1.19 SE). Mean of Group B was 47.05 (47.05+/-4.05 SE) whereas normal controls displayed a mean of 48.66 (48.66+/-2.94 SE). Group A was significantly different from control (p < 0.00001). Group A and Group B were also significant different (p < 0.001) whereas control was not significantly different from Group B (p > 0.75). CONCLUSIONS: Patients with metastatic adenocarcinoma of prostate, as a group, have a significantly lower Leydig cell density than patients without metastasis or patients without PC in compatible age groups. The hormonal relationship between this observation is however unknown. One possible explanation is that PC subpopulation with metastatic potential may require different level of endogenous androgen or are androgen-independent.     

Immunohistochemical staining of endomorphin 1 and 2 in the immune cells of the spleen

Endomorphin 1 (EM-1) and EM-2 have been widely reported in the cells of the central nervous system (CNS) but limited research has been done regarding their distribution in the peripheral system. The occurrence of EM-1 and -2 in the spleen as measured by RIA and their ability to mediate immune function imply a role for EMs in this area. The current study examines the localization of EM-1 and -2 in the immune cells of the spleen of male and female rats via an immunohistochemical procedure. In both genders, EM-1 and -2 immunoreactive staining was predominantly present in macrophages and B cells with minimal EM immunoreactive staining in T cells. This is the first evidence of a differential distribution of EM-1 and -2 in cells of the immune system.     

Production and significance of TGF-beta in AT-3 metastatic cell line established from the Dunning rat prostatic adenocarcinoma

A colony formation assay using NRK-49F cells revealed that a metastatic cell line, AT-3, established from the Dunning prostatic carcinoma could produce TGF-beta in a latent form. TGF-beta at a concentration as low as 0.05 ng/ml either stimulated the attachment or detachment of AT-3 cells depending on the kind of culture media. Acid extracts from conditioned medium (5 micrograms/ml) showed the activity comparable to that of TGF-beta (5 ng/ml). The detached cells were able to grow in suspension. TGF-beta (0.1 ng/ml) could also stimulate the growth of MC3T3-El osteoblasts established from mouse calvaria. These results suggest that TGF-beta is a key growth factor for osteoblastic bony metastasis of prostate cancer.     

Polyclonal carcinoembryonic antigen staining in the cytologic differential diagnosis of primary and metastatic hepatic malignancies.

In order to assess the utility of immunocytochemical staining of bile canaliculi with a polyclonal antiserum to carcinoembryonic antigen (pCEA) in the differentiation of primary hepatocellular carcinomas from metastatic malignancies, pCEA staining was performed on fine needle aspiration specimens from hepatic lesions in 60 patients. The original cytologic diagnoses were hepatocellular carcinoma in 22 patients, metastatic neoplasm or cholangiocarcinoma in 27 patients and benign hepatocytes in 11 cases. The cytologic diagnoses of malignancy were confirmed by surgical excision, autopsy or clinical investigations in 82% of the patients. Follow-up data, supported by pCEA staining, reversed the original cytologic diagnosis in three cases. Bile canalicular pCEA staining was identified in 18 of 22 cases of hepatocellular carcinoma and in all 11 benign hepatocellular aspirates. All 27 cases of metastatic malignancy or cholangiocarcinoma were negative for canalicular pCEA staining, although 11 cases exhibited cytoplasmic staining. Interpretation of pCEA staining was not affected by the intermingling of malignant cells and benign hepatocytes. Predictive values were 100% for a positive test and 87% for a negative test. These findings indicate that staining with pCEA antiserum is a useful adjunct in the differential cytologic diagnosis of malignant hepatic lesions.     

Fine needle aspiration cytology of prostatic adenocarcinoma metastatic to the lung confirmed by the immunoperoxidase technique

Two cases of prostatic adenocarcinoma metastatic to the lung are presented. The prostatic origin of the metastatic carcinomas was confirmed by immunocytochemical demonstration of prostate-specific antigen and prostatic acid phosphatase in cellular samples obtained by fine needle aspiration of the pulmonary lesions.     

Computer-assisted grading of adenocarcinoma in prostatic aspirates

Conventional cytologic grading of fine needle aspirates of prostatic adenocarcinoma has been shown neither to be reproducible nor to correlate well with histologic grading. This study developed a tumor grade classification based on computerized cytomorphometric features and compared the results to conventional grading of companion tissue sections. The image analysis system evaluated architectural features of the aspirates (mainly cell cluster features and interrelationships) as well as nuclear features. Thirty-five prostatic adenocarcinomas (8 well, 19 moderately and 8 poorly differentiated) were evaluated. Discriminant functions based on data collected at medium and high resolution distinguished between aspirates from low-grade (well-differentiated) and high-grade (poorly differentiated) adenocarcinomas with 81% accuracy. Moderately differentiated cancers could not be classified as a distinct group. This study suggests that accurate grading of prostatic adenocarcinoma in fine needle aspirate smears requires the evaluation of medium-resolution features related to specimen cellularity and uniformity or crowding of cell clusters as well as of high-resolution features of nuclear area, perimeter and coarseness of chromatin texture. These findings are compared to those of other schemes for the cytologic grading of prostatic aspirates.     

Differential expression of metabolic genes in tumor and stromal components of primary and metastatic loci in pancreatic adenocarcinoma

Background

Pancreatic cancer is the fourth leading cause of cancer related deaths in the United States with a five-year survival rate of 6%. It is characterized by extremely aggressive tumor growth rate and high incidence of metastasis. One of the most common and profound biochemical phenotypes of animal and human cancer cells is their ability to metabolize glucose at high rates, even under aerobic conditions. However, the contribution of metabolic interrelationships between tumor cells and cells of the surrounding microenvironment to the progression of cancer is not well understood. We evaluated differential expression of metabolic genes and, hence, metabolic pathways in primary tumor and metastases of patients with pancreatic adenocarcinoma.

Methods and Findings

We analyzed the metabolic gene (those involved in glycolysis, tri-carboxylic acid pathway, pentose-phosphate pathway and fatty acid metabolism) expression profiles of primary and metastatic lesions from pancreatic cancer patients by gene expression arrays. We observed two principal results: genes that were upregulated in primary and most of the metastatic lesions; and genes that were upregulated only in specific metastatic lesions in a site-specific manner. Immunohistochemical (IHC) analyses of several metabolic gene products confirmed the gene expression patterns at the protein level. The IHC analyses also revealed differential tumor and stromal expression patterns of metabolic enzymes that were correlated with the metastasis sites.

Conclusions

Here, we present the first comprehensive studies that establish differential metabolic status of tumor and stromal components and elevation of aerobic glycolysis gene expression in pancreatic cancer.     

Allelotyping of human prostatic adenocarcinoma.

Allelotyping (using at least one probe detecting a restriction fragment length polymorphism on each chromosomal arm, with the exception of the short arms of the acrocentric chromosomes), showed loss of genetic information in 11 of 18 prostate adenocarcinoma specimens analyzed (61%). Frequent allelic deletions were detected on the long arm of chromosome 16 (6 of 10 informative cases, 60%), on the short arm of chromosome 8 (3 of 6 informative cases, 50%), and on the short and/or the long arms of chromosome 10 (6 of 11 informative cases (10p), 55% and 4 of 13 informative cases (10q), 30%, respectively). No losses of alleles were detected in any case unless at least one of the chromosomes 8, 10, or 16 also showed deletions. The long arm of chromosome 18 also showed a high frequency of allelic deletions (3 of 7 informative cases, 43%). Allelic deletions on the following chromosomes were detected at lower frequencies: chromosomes 2, 3, 7, 12, 13, 17, 22, and XY. Tumors with allelic deletions on more than one chromosome had a higher histological malignancy grade. Tumors from patients with advanced disease all showed allelic deletions.     

Immunohistochemical staining of post-mortem adult human brain sections

One of the challenges for modern neuroscience is to understand the basis of coordinated neuronal function and networking in the human brain. Some of these questions can be addressed using low- and high-resolution imaging techniques on post-mortem human brain tissue. We have established a versatile protocol for fixation of post-mortem adult human brain tissue, storage of the tissue in a human brain bank, and immunohistochemical analysis in order to understand human brain functions in normal controls and in neuropathological conditions. The brains are fixed by perfusion through the internal carotid and basilar arteries to enhance the penetration of fixative throughout the brain, then blocked, postfixed, cryoprotected, snap-frozen and stored at -80 degrees C. Sections are processed for immunohistochemical single- or double-label staining and conventional-, electron- or confocal laser scanning-microscopy analysis. The results gained using this tissue and protocol are vital for determining the localization of neurochemicals throughout the human brain and to document the changes that occur in neurological diseases.     

Immunohistochemical analysis and comparison of napsin A,TTF1, SPA and CK7 expression in primary lung adenocarcinoma

We investigated the expression of napsin A, TTF1, SPA and CK7 in primary lung adenocarcinoma (ADC) and compared antibody specificity and sensitivity. We used immunohistochemistry (IHC) using antibodies directed against napsin A, TTF1, SPA, and CK7 on cancer tissue from 454 patients diagnosed with lung cancer. In primary lung ADC, napsin A exhibited a sensitivity of 87.25% and specificity of 95.02%, TTF1 exhibited a sensitivity of 80.72% and specificity of 90.48%, SPA exhibited a sensitivity of 60.46% and specificity of 94.87% and CK7 exhibited a sensitivity of 100% and specificity of 80.31%. The order of sensitivity of the four indicators was CK7 > napsin A > TTF1 > SPA, and the order of specificity was napsin A > SPA > TTF1 > CK7. We can select effective indicators for diagnostic and differential diagnostic purposes based on differences in sensitivity and specificity.     

Herpesviruses in metastatic Lucké renal adenocarcinoma

The etiologic agent of the renal adenocarcinoma of leopard frogs, Rana pipiens, is the Lucké tumor herpesvirus (LTHV). The virus is easily detected with thin section electron microscopy in primary tumors of frogs which have been exposed to a cold environment. Several spontaneous metastatic nodules and a large primary tumor were detected at autopsy of a frog which had been maintained at 4 degrees C for 73 days. LTHV was found not only in the primary tumor, as previously reported, but also was present in metastatic tumor cells in the liver, fat body, and bladder. The presence of LTHV in metastatic cells demonstrates that the differentiated state of primary Lucké tumor cells is retained in its metastatic colonies even at the fine structure level revealed by electron microscopy.     

Non-bacterial thrombotic endocarditis in metastatic caecal adenocarcinoma

A 34-year-old woman was admitted because of headache, blurred vision, apraxia and abnormal feeling in the right arm. Examination revealed finger petechiae. Infection parameters were low. Magnetic resonance imaging of the brain showed multiple cortical and white matter lesions.     

Colon carcinoma metastatic to the lung. Cytologic manifestations and distinction from primary pulmonary adenocarcinoma.

Based upon a review of cytologic specimens obtained from 46 patients, the morphologic characteristics of metastatic colon carcinoma and primary pulmonary adenocarcinoma were compared and contrasted. A monoclonal antibody (D-14) reported to be helpful in identifying colorectal carcinoma was also evaluated. Colon brushings and washings from 10 patients with colonic carcinoma and a variety of respiratory tract cytology samples from 16 and 20 patients with metastatic colonic and primary pulmonary adenocarcinomas, respectively, formed the basis of this review. The presence of well-formed glands and dirty necrosis was significantly more characteristic of colon carcinoma. Other cytologic features and D-14 staining were not sufficiently distinctive to allow a separation from primary pulmonary adenocarcinoma.     

Expression of prostate specific membrane antigen (PSMA) in prostatic adenocarcinoma and prostatic intraepithelial neoplasia

The prostatic membrane antigen (PSMA) is a protein that is expressed in the prostatic epithelium. We studied the expression of PSMA in a series of 55 patients with different stages of prostate cancer and we compared the PSMA staining in prostate cancer cells, in high-grade prostatic intraepithelial neoplasia (PIN) and in histologically benign prostatic epithelium for the same specimen. For this purpose archival paraffin-embedded specimens were studied by immunohistochemistry with a monoclonal antibody 7E11-C5.3 against PSMA using the streptavidin-biotin method. The mean percentage of PSMA immunoreactivity was 56.67% in prostate cancer (CaP) cells, and 48.6% in PIN cells, which was significantly higher than benign-appearing prostatic epithelium (5.72%) (for each pair, p<0.001). PSMA expression was greater in CaP with a higher Gleason score (p=0.01), but no relationship was found with serum PSA value. We conclude that PSMA overexpression is detected in high-grade PIN and is associated with a higher Gleason score of prostate cancer. It is a potential marker for studying carcinogenesis and progression of prostate cancer.     

Characterizing "blue blobs". Immunohistochemical staining and ultrastructural study

OBJECTIVE: To investigate the nature and origin of "blue blobs" (Bbs) in atrophic Pap smears in postmenopausal women and to study their clinical significance. STUDY DESIGN: A retrospective study of 412 atrophic Pap smears from postmenopausal women was done to detect the presence of Bbs. The smears from 24 cases showing Bbs were further studied to evaluate the nature of the Bbs with special stains, immunohistochemistry and electron microscopy. RESULTS: Bbs showed a heterogeneous morphology, with variable numbers and staining intensity. The diameter of Bbs was approximately equivalent to that of a parabasal/intermediate squamous cell. Special stains showed Bbs to be positive for periodic acid-Schiff and methyl green pyronin and negative for mucicarmine and calcium. Immunohistochemistry revealed Bbs to be positive for cytokeratin, epithelial membrane antigen and carcinoembryonic antigen and negative for vimentin and muscle-specific actin. Some Bbs had residual ghost nuclear shadows. Electron microscopy revealed cellular skeletons with residual tonofilaments enmeshed within a loose cytoskeleton matrix and nuclei with variable degrees of degeneration. CONCLUSION: Special stains, immunohistochemistry and electron microscopy indicated that Bbs represent parabasal/intermediate squamous cells exhibiting various degree of degeneration. In general, Bbs appear to be of no clinical significance except as a source of potential diagnostic error.