[show abstract][hide abstract] ABSTRACT: Pancreatic carcinoma is one of the cancers with the worse prognosis, thus any therapeutic improvement is imperative. Cytotoxic LH-RH analog, AN-152 (proprietary designation, AEZS-108), consisting of doxorubicin (DOX) conjugated to D-Lys6LH-RH, is now in clinical trials for targeted therapy of several sex hormone-dependent tumors that express LH-RH receptors. We investigated LH-RH receptors in human pancreatic carcinoma and the effects of AN-152 (AEZS-108) on experimental pancreatic cancers. We determined LH-RH receptor presence in human pancreatic cancer samples by immunohistochemistry and, in three human pancreatic cancer lines (SW-1990, Panc-1 and CFPAC-1), by binding assays and Western blotting. The effects of the cytotoxic LH-RH analog were investigated on growth of these same cancer lines xenografted into nude mice. We also analyzed differences between the antitumor effects of the cytotoxic analog and its cytotoxic radical alone, doxorubicin (DOX), on the expression of cancer-related genes by PCR arrays. LH-RH receptors were expressed in two randomly selected surgically removed human pancreatic cancer samples and in all three cancer lines. Cytotoxic LH-RH analogs powerfully inhibited growth of all three tumor lines in nude mice; AN-152 was significantly stronger than DOX on Panc-1 and CFPAC-1 cancers. PCR array showed that cytotoxic LH-RH analog AN-152 affected the expression of genes associated with cellular migration, invasion, metastasis and angiogenesis more favorably than DOX, however the changes in gene expression varied considerably among the three cancer lines. Cytotoxic LH-RH analog, AEZS-108, may be a useful agent for the treatment of LH-RH receptor positive advanced pancreatic carcinoma.
[show abstract][hide abstract] ABSTRACT: Glioblastoma multiforme is the most frequent tumor of the central nervous system in adults and has a dismal clinical outcome, which necessitates the development of new therapeutic approaches. We investigated in vivo the action of the targeted cytotoxic analog of luteinizing hormone releasing hormone, AN-152 (AEZS-108) in nude mice (Ncr nu/nu strain) bearing xenotransplanted U-87 MG glioblastoma tumors. We evaluated in vitro the expression of LHRH receptors, proliferation, apoptosis and the release of oncogenic and tumor suppressor cytokines. Clinical and U-87 MG samples of glioblastoma tumors expressed LHRH receptors. Treatment of nude mice with AN-152, once a week at an intravenous dose of 413 nmol/20g, for six weeks resulted in 76 % reduction in tumor growth. AN-152 nearly completely abolished tumor progression and elicited remarkable apoptosis in vitro. Genomic (RT-PCR) and proteomic (ELISA, Western blot) studies revealed that AN-152 activated apoptosis, as reflected by the changes in p53 and its regulators and substrates, inhibited cell growth, and elicited changes in intermediary filament pattern. AN-152 similarly reestablished contact regulation as demonstrated by expression of adhesion molecules and inhibited vascularization, as reflected by the transcription of angiogenic factors. Our findings suggest that targeted cytotoxic analog AN-152 (AEZS-108) should be considered for a treatment of glioblastomas.
[show abstract][hide abstract] ABSTRACT: Mammaglobin A (MG-A) is purportedly useful for detecting metastatic carcinomas suspected to be of breast origin and has been advocated as a useful marker of micrometastasis in sentinel lymph nodes and minimal residual tumor in bone marrow. Little is known about its expression frequency in histologic subtypes of breast cancer. Excisional biopsy specimens from 1,079 untreated invasive mammary carcinomas were evaluated for immunohistochemical expression of MG-A. In addition to estrogen (ER) and progesterone receptors (PR) and HER2, staining for p63 and HLA-DR was used to further characterize histologic subtypes. Of the carcinomas, 36 were classified as metaplastic (based on morphologic features, ER-/PR-/HER2-, p63+), 38 as medullary (ER-/PR-/HER2-, HLA-DR+), and 1,005 as ductal, no special type (NST). All metaplastic and medullary carcinomas were negative for MG-A. Of 1,005 ductal carcinomas, NST, 492 (49.0%) were MG-A+, 62.0% with a reaction in fewer than 25% of the cells. MG-A immunohistochemical studies failed to detect all medullary and metaplastic cancers and more than 50% of ductal carcinomas, NST. In two thirds of MG-A+ ductal carcinomas, the reaction was only focal and usually in a minority of cells. These findings suggest that MG-A has limited value in identifying the mammary origin of carcinomas, particularly in small biopsy specimens used to detect metastasis or minimal residual disease.
American Journal of Clinical Pathology 05/2012; 137(5):747-52. · 2.88 Impact Factor
[show abstract][hide abstract] ABSTRACT: Experimental data indicate that antagonists of growth hormone-releasing hormone (GHRH) could be used clinically in disorders characterized by excessive GHRH/growth hormone (GH) secretion, but direct evidence for the effectiveness of GHRH antagonists on human pituitary tissue is still lacking. In this study, we investigated the inhibitory effect of our GHRH antagonists MZ-4-71 and JV-1-36 and the somatostatin (SST) analog RC-160 on superfused pituitary cells obtained from a human GH-secreting adenoma. Using Western blot analysis and immunohistochemistry, we demonstrated profuse expression of the GHRH receptor and its major splice variant SV1 and an increase in the expression of Gsa protein in the adenoma tissue. Exposure of the tumor cells to exogenous pulses of GHRH induced definite GH responses, causing a 3- to 5-fold elevation of the basal GH level. The antagonists MZ-4-71 and JV-1-36 did not alter basal GH secretion, indicating that the adenoma cells did not secrete GHRH in an autocrine manner. However, both antagonists prevented the stimulatory effect of exogenous GHRH. Similarly to the GHRH antagonists, neither SST-14 nor the SST analog RC-160 had an effect on the basal GH secretion of the tumor cells, but both peptides inhibited the stimulatory effect of exogenous GHRH, with RC-160 being more potent than SST. Our study provides direct evidence for the effectiveness of potent GHRH antagonists such as MZ-4-71 and JV-1-36 on human pituitary GH-secreting adenoma tissue and strongly suggests that these drugs could be used for therapy of GHRH-associated forms of acromegaly, particularly for those patients in whom surgery fails or is not an option.
[show abstract][hide abstract] ABSTRACT: Benign prostatic hyperplasia often affects aging men. Antagonists of the neuropeptide growth hormone-releasing hormone reduced prostate weight in an androgen induced benign prostatic hyperplasia model in rats. Luteinizing hormone-releasing hormone antagonists also produce marked, protracted improvement in lower urinary tract symptoms, reduced prostate volume and an increased urinary peak flow rate in men with benign prostatic hyperplasia. We investigated the influence of a combination of antagonists of growth hormone-releasing hormone and luteinizing hormone-releasing hormone on animal models of benign prostatic hyperplasia.
We evaluated the effects of the growth hormone-releasing hormone antagonist JMR-132, given at a dose of 40 μg daily, the luteinizing hormone-releasing hormone antagonist cetrorelix, given at a dose of 0.625 mg/kg, and their combination on testosterone induced benign prostatic hyperplasia in adult male Wistar rats in vivo. Prostate tissue was examined biochemically and histologically. Serum levels of growth hormone, luteinizing hormone, insulin-like growth factor-1, dihydrotestosterone and prostate specific antigen were determined.
Marked shrinkage of the rat prostate (30.3%) occurred in response to the combination of growth hormone-releasing hormone and luteinizing hormone-releasing hormone antagonists (p<0.01). The combination strongly decreased prostatic prostate specific antigen, 6-transmembrane epithelial antigen of the prostate, interleukin-1β, nuclear factor-κβ and cyclooxygenase-2, and decreased serum prostate specific antigen.
A combination of growth hormone-releasing hormone antagonist with luteinizing hormone-releasing hormone antagonist potentiated a reduction in prostate weight in an experimental benign prostatic hyperplasia model. Results suggest that this shrinkage in prostate volume was induced by the direct inhibitory effects of growth hormone-releasing hormone and luteinizing hormone-releasing hormone antagonists exerted through their respective prostatic receptors. These findings suggest that growth hormone-releasing hormone antagonists and/or their combination with luteinizing hormone-releasing hormone antagonists should be considered for further development as therapy for benign prostatic hyperplasia.
The Journal of urology 02/2012; 187(4):1498-504. · 4.02 Impact Factor
[show abstract][hide abstract] ABSTRACT: The management of castration-resistant prostate cancer (CRPC) presents a clinical challenge because of limitations in efficacy of current therapies. Novel therapeutic strategies for the treatment of CRPC are needed. Antagonists of hypothalamic growth hormone-releasing hormone (GHRH) inhibit growth of various malignancies, including androgen-dependent and independent prostate cancer, by suppressing diverse tumoral growth factors, especially GHRH itself, which acts as a potent autocrine/paracrine growth factor in many tumors. We evaluated the effects of the GHRH antagonist, JMR-132, on PC-3 human androgen-independent prostate cancer cells in vitro and in vivo. JMR-132 suppressed the proliferation of PC-3 cells in vitro in a dose-dependent manner and significantly inhibited growth of PC-3 tumors by 61% (P < 0.05). The expression of GHRH, GHRH receptors, and their main splice variant, SV1, in PC-3 cells and tumor xenografts was demonstrated by RT-PCR and Western blot. The content of GHRH protein in PC-3 xenografts was lowered markedly, by 66.3% (P < 0.01), after treatment with JMR-132. GHRH induced a significant increase in levels of ERK, but JMR-132 abolished this outcome. Our findings indicate that inhibition of PC-3 prostate cancer by JMR-132 involves inactivation of Akt and ERK. The inhibitory effect produced by GHRH antagonist can result in part from inactivation of the PI3K/Akt/mammalian target of rapamycin and Raf/MEK/ERK pathways and from the reduction in GHRH produced by cancer cells. Our findings support the role of GHRH as an autocrine growth factor in prostate cancer and suggest that antagonists of GHRH should be considered for further development as therapy for CRPC.
Proceedings of the National Academy of Sciences 01/2012; 109(5):1655-60. · 9.74 Impact Factor
[show abstract][hide abstract] ABSTRACT: The majority of men will develop symptoms of benign prostatic hyperplasia (BPH) after 70 years of age. Various studies indicate that antagonists of LHRH, such as cetrorelix, exert direct inhibitory effects on BPH mediated by specific LHRH receptors. Our aim was to investigate the mRNA for LHRH and LHRH receptors and the expression of LHRH receptors in specimens of human BPH.
The expression of mRNA for LHRH (n=35) and LHRH receptors (n=55) was investigated by RT-PCR in surgical specimens of BPH, using specific primers. The characteristics of binding sites for LHRH on 20 samples were determined by ligand competition assays. The LHRH receptor expression was also examined in 64 BPH specimens by immunohistochemistry.
PCR products for LHRH were found in 18 of 35 (51%) BPH tissues and mRNA for LHRH receptors was detected in 39 of 55 (71%) BPH specimens. Eighteen of 20 (90%) samples showed a single class of high affinity binding sites for [D-Trp(6) ]LHRH with a mean K(d) of 4.04 nM and a mean B(max) of 527.6 fmol/mg membrane protein. LHRH antagonist cetrorelix showed high affinity binding to LHRH receptors in BPH. Positive immunohistochemical reaction for LHRH receptors was present in 42 of 64 (67%) BPH specimens.
A high incidence of LHRH receptors in BPH supports the use of LHRH antagonists such as cetrorelix, for treatment of patients with lower urinary tract symptoms from BPH.
The Prostate 04/2011; 71(5):445-52. · 3.84 Impact Factor
[show abstract][hide abstract] ABSTRACT: Growth hormone-releasing hormone (GHRH), a hypothalamic polypeptide, acts as a potent autocrine/paracrine growth factor in many cancers. Benign prostatic hyperplasia (BPH) is a pathologic proliferation of prostatic glandular and stromal tissues; a variety of growth factors and inflammatory processes are inculpated in its pathogenesis. Previously we showed that potent synthetic antagonists of GHRH strongly inhibit the growth of diverse experimental human tumors including prostate cancer by suppressing various tumoral growth factors. The influence of GHRH antagonists on animal models of BPH has not been investigated. We evaluated the effects of the GHRH antagonists JMR-132 given at doses of 40 μg/d, MIA-313 at 20 μg/d, and MIA-459 at 20 μg/d in testosterone-induced BPH in Wistar rats. Reduction of prostate weights was observed after 6 wk of treatment with GHRH antagonists: a 17.8% decrease with JMR-132 treatment; a 17.0% decline with MIA-313 treatment; and a 21.4% reduction with MIA-459 treatment (P < 0.05 for all). We quantified transcript levels of genes related to growth factors, inflammatory cytokines, and signal transduction and identified significant changes in the expression of more than 80 genes (P < 0.05). Significant reductions in protein levels of IL-1β, NF-κβ/p65, and cyclooxygenase-2 (COX-2) also were observed after treatment with a GHRH antagonist. We conclude that GHRH antagonists can lower prostate weight in experimental BPH. This reduction is caused by the direct inhibitory effects of GHRH antagonists exerted through prostatic GHRH receptors. This study sheds light on the mechanism of action of GHRH antagonists in BPH and suggests that GHRH antagonists should be considered for further development as therapy for BPH.
Proceedings of the National Academy of Sciences 02/2011; 108(9):3755-60. · 9.74 Impact Factor
[show abstract][hide abstract] ABSTRACT: Enhanced expression of aldehyde dehydrogenase 1 (ALDH1) and phenotypic markers (CD44(+)/CD24(-)) in stem cells from breast tumors has been reported. This study was undertaken to monitor expression of these markers in cells from body cavity fluids of female patients suspected to have a malignancy.
Cells from peritoneal and pleural fluids of 100 female patients were examined by diagnostic cytology and analyzed by laser flow cytometry for enhanced ALDH1 expression. Cells from 36 body cavity fluids with ALDH1(bright) fluorescence were then analyzed for the expression of CD44 and CD24 markers.
In samples positive for malignancy, ALDH1(bright) cells with both SSC(low) and SSC(high) were seen. In 15 body cavity fluids positive for malignancy, the percentage of ALDH1(bright) cells ranged from 0.26 to 6.34% of the total cells. The percentage of ALDH1(bright) cells with CD44(+)/CD24(-) expression in these samples ranged from 0.02 to 3.66%. ALDH1(bright) cells with CD44(+)/CD24(-) expression were also present in body cavity fluids of patients in whom diagnostic cytology could not detect any malignancy. However, the percentage of ALDH1(bright) and CD44(+)/CD24(-) cells amongst the 21 body cavity fluids with negative cytology was lower than that of samples with malignancy.
Expression of ALDH1(bright) and the CD44(+)/CD24(-) phenotype in body cavity fluids in which diagnostic cytology could not find any malignant cells suggests that this phenotype may not be restricted to the putative breast tumor stem cells. It is possible that only subsets of cells with this phenotype are the putative breast tumor stem cells.
Cytometry Part B Clinical Cytometry 05/2010; 78(3):176-82. · 2.23 Impact Factor
[show abstract][hide abstract] ABSTRACT: Primary enteric-type adenocarcinoma of the urinary bladder is relatively uncommon. We present our experience with 109 pure, non-urachal cases—the largest series to date. This work was undertaken with the aim of describing the immunohistochemical features of adenocarcinoma of the urinary bladder associated with schistosomiasis, illustrating their histologic and immunohistochemical similarities to colorectal carcinomas. Partial or total cystectomy specimens from a cohort of Egyptian and American patients with the diagnosis of primary adenocarcinoma of the urinary bladder (109 cases) were reviewed. Paraffin sections of each tumour were stained using the labelled streptavidin–biotin method using antibodies cytokeratin 20, cytokeratin 7, CDX2, MLH1, and villin. Clinical follow-up was available for at least 36 months. An enteric (colonic) morphology was seen in most tumours; some with signet ring cells or mucinous elements. Five tumours were composed predominantly of signet ring cells and two demonstrated a pure mucinous morphology. In cases where adjacent normal mucosa was present, 23% showed either colonic metaplasia or intestinal-type cystitis glandularis. Furthermore, 24% of enteric-type adenocarcinomas had associated villous or tubulovillous adenomas with or without dysplasia. Cytokeratin 20 was expressed by 90%, cytokeratin 7 by 17%, CDX2 by 100% and villin and MLH1 by 76% and 48% of tumours respectively. The majority of tumours presented with an advanced stage and followed an aggressive clinical course.In conclusion, primary non-urachal enteric-type adenocarcinoma of the urinary bladder is morphologically and immunophenotypically similar – if not identical – to colonic adenocarcinoma. The frequent association of enteric carcinomas of the urinary bladder with intestinal metaplasia and/or colonic-type adenomas with dysplasia suggests possible carcinogenetic pathways similar to that observed in colorectal carcinomas.
Journal of Advanced Research. 01/2010; 1(2):151-156.
[show abstract][hide abstract] ABSTRACT: Diagnostic cytology based on the examination of cells from body cavity fluids misses approximately 50% of patients with a proven malignancy. In an earlier study, we used immunohistochemical detection of epithelial membrane antigen expression with flow cytometric detection of DNA aneuploidy to reduce the number of false negatives. In the present study, we have combined DNA flow cytometry with flow cytometric detection of marker expression to analyze cells from body cavity fluids. Seventy-nine specimens of ascites and pleural fluids were analyzed by diagnostic cytology, DNA flow cytometry, and for the expression of the following markers: Ber-EP4, progesterone (PR), MUC4, and thyroid transcription factor-1 (TTF-1). DNA index of equal to or greater than 1.2 was seen in 33/79 (41.7%) of the samples. Statistical analysis of 79 samples in which data from cytology, DNA aneuploidy, and expression of at least one of the markers was available showed that by combining data from positive marker expression with that of aneuploidy, the sensitivity was increased from 58.5 to 100%. In contrast, out of the 38 samples designated as non-malignant by diagnostic cytology, nine had aneuploid DNA content and 16 of the diploid samples had a positive marker expression. Specificity was reduced from 74.7 to 31.6% due to the presence of aneuploidy and marker expression in these samples. ALDH1(pos)/CD44(pos)/CD24(neg) expression has been reported to be associated with human breast tumor stem cells. Some of our samples had cells with this phenotype. Flow cytometry offers the advantage of rapid multiparametric analysis of DNA aneuploidy and marker expression in cells from body cavity fluids based on the analysis of a large number of cells without observer bias. By further developing the use of specific markers and aneuploidy, it may be possible to refine flow cytometric analysis for rapid detection of malignant cells in body cavity fluids.
Cytometry Part A 11/2009; 77(2):132-43. · 3.71 Impact Factor
[show abstract][hide abstract] ABSTRACT: Immunohistochemistry for estrogen receptor may be used to distinguish metastatic breast cancers from adenocarcinomas of other sites, including those of the lung. The estrogen receptor exists as 2 subtypes, alpha and beta. Estrogen receptor alpha is the predominant subtype expressed by more than two-thirds of human breast cancers. Adenocarcinomas of lung origin may also express estrogen receptor, primarily the beta subtype. Human estrogen receptor alpha is highly homologous to estrogen receptor beta and consequently, antibodies used to detect estrogen receptor alpha in breast carcinomas may detect estrogen receptor beta in pulmonary adenocarcinomas. We investigated the immunohistochemical expression of estrogen receptor in proven primary lung adenocarcinomas using 3 anti-estrogen receptor alpha antibodies: mouse monoclonal 1D5, 6F11, and rabbit monoclonal SP1.
Ninety-two pulmonary adenocarcinomas (53 women and 39 men) confirmed by clinical presentation and positive immunohistochemistry for thyroid transcription factor-1 (TTF-1) were included in this study. There were 19 incisional biopsies and 73 excisional specimens. Immunohistochemistry for estrogen receptor using antibodies 1D5, 6F11, and SP1 was performed on formalin-fixed, paraffin-embedded tissue following antigen retrieval. Any nuclear reactivity for estrogen receptor was considered a positive result.
Focal positive nuclear reaction for estrogen receptor was detected in 7 (7.6%) cases of primary pulmonary adenocarcinoma using antibody 1D5, 13 (14.1%) using 6F11, and 25 (27.2%) using SP1. The differences in reactivity for estrogen receptor in pulmonary adenocarcinomas between SP1 and 1D5, and between SP1 and 6F11 were statistically significant (P<0.001). Positive cases showed only a focal pattern of staining with each of the 3 antibodies. There was no significant difference in reactivity for estrogen receptor in pulmonary adenocarcinomas of men and women. Positive staining was highest in nonmucinous bronchioloalveolar adenocarcinomas for all of the antibodies, and for SP1, variation by histologic subtype was significant (P<0.001).
SP1 has a significantly higher detection rate for the expression of estrogen receptor in pulmonary adenocarcinomas when compared with either 1D5 or 6F11. Caution should therefore be exercised in the use of this antibody alone in distinguishing a metastatic breast from a primary pulmonary adenocarcinoma.
Applied immunohistochemistry & molecular morphology: AIMM / official publication of the Society for Applied Immunohistochemistry 10/2009; 18(2):137-41. · 1.63 Impact Factor
[show abstract][hide abstract] ABSTRACT: We evaluated the estrogen receptor (ER) phenotype of recurrent and/or metastatic breast cancers and compared it with the ER status of the primary tumor in 278 cases. All patients had undergone surgical excision of the primary tumor, followed by observation only or treatment modalities that included radiation and/or chemotherapy or hormonal therapy. The immunohistochemical expression of ER was evaluated by using monoclonal antibody ER-1D5 and heat-induced antigen retrieval. At diagnosis, 165 patients had locoregional disease and 7 had distant metastases. Local recurrences and/or distant metastases occurred from 2 months to 21 years after the diagnosis of the primary tumor. Overall, 159 primary tumors (57.2%) were positive for ER. In 269 cases (96.8%), the ER status of the primary and metastatic tumors was the same. In 9 patients with ER+ primary tumors, the metastases were ER-. There were no ER- primary tumors with ER+ metastases. The time to distant metastasis was significantly longer for ER+ tumors. The immunohistochemically determined ER phenotype of primary breast cancers remains stable in most recurrent and metastatic disease.
American Journal of Clinical Pathology 01/2009; 130(6):879-82. · 2.88 Impact Factor
[show abstract][hide abstract] ABSTRACT: Currently, surgeons have to wait for at least 1 day to receive the pathology report of a biopsy or other surgical excision. This delay is mandated by the overnight tissue-processing methods that have been in use for more than a century. Patient anxiety and delay in treatment are consequences of this practice. Here we report the impact of a tissue-processing system on the turnaround time of surgical pathology reporting and its potential effect on overall patient management. This technique provides the feasibility for performing molecular assays on the same sample used for pathologic diagnosis.
Biopsies and other surgically removed specimens from patients treated at the University of Miami, Jackson Memorial Hospital during calendar year 2005 were processed by an automated, microwave-assisted rapid tissue-processing method. Turnaround time for surgical pathology reports was calculated and compared with that of year 1996, the last year before the new technology was phased in.
Total tissue-processing time was reduced from 8 to 10 hours to 67 minutes, resulting in the availability of slides in less than 3 hours. In 80% of the patients, diagnoses were reported on the same day they were received in the laboratory. The 1-day turnaround for the reports in 1996 was < 1%. Histology of rapidly processed tissues and their histochemical and immunohistochemical properties were comparable with those of the traditionally prepared material.
The rapid turnaround capability of the new tissue-processing system has allowed the pathology laboratory to render the final report in the majority of specimens on the day they are received. The feasibility of preserving macromolecules in the same clinical samples used for diagnosis is a timely advantage in the era of molecular medicine.
Journal of the American College of Surgeons 10/2008; 207(3):320-5. · 4.50 Impact Factor
[show abstract][hide abstract] ABSTRACT: "The value of a clinical test should be assessed in the overall context of disease management." The ultimate goal of an assay for detection of estrogen receptor (ER) content in breast cancer tissue is to identify patients who will or will not benefit from endocrine therapy. In the past 2 decades, scenarios for ER testing of patient samples have shifted from tissue homogenate-based, biochemical ligand-binding assays to the more practical and clinically relevant slide-based immunohistochemical methods. Although the superiority of the predictive value of ER-immunohistochemistry (ER-IHC) over ligand-binding techniques has been established to everyone's satisfaction, there remains the controversial issue of quantitation of immunohistochemical results. The assumption that ER-IHC should be quantitative stems largely from the fact that the old biochemical assay results were numerical. Seasoned immunohistochemists, nevertheless, know that IHC of routinely fixed and processed tissue does not yield itself to accurate quantitation of results, even when performed by well-qualified laboratories. Furthermore, in the case of ER, immunohistochemical methods only identify a segment or epitope of ER protein that is immunologically reactive with the used antibody. Hence, as it is, an immunohistochemical technique gives no information about the functional status of ER molecule, and/or that of the complex downstream ER pathways. This may be one of the reasons why one-third of patients with ER-positive breast cancers initially, and another one-third eventually, do not respond to endocrine treatment modalities. In this review, I attempt to present an argument that is based on our current information; quantitation of ER-IHC is neither technically reliable nor clinically relevant.
Applied immunohistochemistry & molecular morphology: AIMM / official publication of the Society for Applied Immunohistochemistry 04/2008; 16(2):105-7. · 1.63 Impact Factor
[show abstract][hide abstract] ABSTRACT: In addition to providing a timely and accurate diagnosis, pathologists routinely provide prognostic and predictive information to assist in the treatment of patients with invasive breast cancer. As our understanding of breast cancer at the molecular and genetic level improves, sophisticated new treatment options have become available to patients. The demonstrated improvements in disease-free and overall survival with the use of trastuzumab (Herceptin) has made HER2 testing a standard of care in the evaluation of patients with breast cancer. Specialized breast centers have accumulated sufficient experience to recognize that HER2 positive tumors tend to be of higher grade and to be estrogen receptor negative, whereas well-differentiated breast cancers rarely are HER2 positive.
To determine whether HER2 testing is necessary in well-differentiated breast cancer, we analyzed the frequency of HER2 positivity among 1,162 cases from 7 major breast centers or commercial laboratories in the United States and Europe.
Well-differentiated breast cancers, defined by either nuclear grading or the Scarff-Bloom-Richardson system, rarely are HER2 positive (mean 1.6%, range 0-2.8%).
Given the low rate of well differentiated HER2 positive tumors, falling within the range reported for false negative IHC tests for HER2, and the absence of published data demonstrating a beneficial effect of trastuzumab therapy in this subset of patients, HER2 testing should not be considered a standard of care for all patients with well-differentiated breast cancer.
Breast Cancer Research and Treatment 02/2008; 112(3):551-6. · 4.47 Impact Factor
[show abstract][hide abstract] ABSTRACT: The potential problems associated with the use of formalin in histology, such as health hazards, degradation of RNA and cross-linking of proteins are well recognized. We describe the utilization of a formalin-free fixation and processing system for tissue detection of two important biopredictors in breast cancer - estrogen receptor and HER2 - at the RNA and protein levels.
Parallel sections of 62 cases of breast cancer were fixed in an alcohol-based molecular fixative and in formalin. Molecular fixative samples were processed by a novel formalin-free microwave-assisted processing system that preserves DNA, RNA and proteins. Formalin-fixed samples were processed using the conventional method. Estrogen receptor was assessed by immunohistochemistry and real-time PCR. HER2 was assessed by immunohistochemistry, FISH, CISH and real-time PCR.
The immunohistochemical reaction for estrogen receptor was similar in molecular- and formalin-fixed samples (Spearman Rank R = 0.83, p < 0.05). Also HER2 result was similar to that of formalin-fixed counterparts after elimination of antigen retrieval step (Spearman Rank R = 0.84, p < 0.05). The result of HER2 amplification by FISH and CISH was identical in the molecular fixative and formalin-fixed samples; although a shorter digestion step was required when using the former fixative. Real-time PCR for both estrogen receptor and HER2 were successful in all of the molecular fixative specimens.
The formalin-free tissue fixation and processing system is a practical platform for evaluation of biomolecular markers in breast cancer and it allows reliable DNA and RNA and protein studies.
[show abstract][hide abstract] ABSTRACT: AimTo determine whether universal molecular fixative (UMFIX), a novel human tissue fixative, could also be used as an animal tissue fixative for histomorphology and a preservative of RNA at subtropical temperatures. Cat, dog, mouse, pigeon, rabbit and rat tissue, as well as ant, beetle, earthworm and lizard were collected. Tissue was fixed in UMFIX for up to a week at room or ambient temperatures, processed and paraffin embedded. Histomorphology and RNA quality were evaluated and compared to formalin-fixed and fresh frozen tissue.