[Tonsillar lymphangiomatous polyp]. / Lymphangiomatöser Tonsillenpolyp.

Lymphangiomatous polyps are rare benign lesions of the tonsils. We report the case of a female patient presenting with dysphagia of 15 years' standing due to such a lesion. After surgical excision the patient remained free of symptoms and experienced no recurrence (follow-up 17 months). The histology was characterized by epitheliotropism of lymphocytes as well as dilated lymphatic channels, blood vessels and edema. Immunohistochemical staining with D2-40 monoclonal antibody and podoplanin confirmed the diagnosis of lymphangiomatous polyps. We discuss the current literature.

[Prostate cancer research. Biomarkers as promising options for optimized diagnosis and treatment]. / Prostatakarzinomforschung. Biomarker als Hoffnung für optimierte Diagnostik und Therapie.

A better understanding of signal transduction and gene regulation during prostate carcinogenesis will allow the development of novel diagnostic and prognostic biomarkers and a better prediction of the individual course of prostate cancer disease. It will also enhance the design and development of specific small molecular components aiming for specific therapies. The research groups in Bonn succeeded in the competition for an endowed professorship supported by the Rudolf Becker Stiftung (German Science Endowment Fund) settled in the"Centrum für integrierte Onkologie" funded by the German Cancer Aid. This should be the perfect breeding ground for future research in the field of prostate cancer.

[Orbital aneurysmal bone cyst in a 2-year-old child]. / Aneurysmatische Knochenzyste im Bereich der Orbita bei einem 2-jährigen Kind.

An aneurysmal bone cyst is a rare tumor-like lesion which can affect any part of the skeleton. It is a disease of childhood and adolescence. Reports of its occurrence on the skull base in children are rare. A 22-month-old male patient was admitted to our ENT department with a sudden protrusion of the right eyeball. Radiologically, a cystic, well-defined and contrast enhanced mass on the medial-cranial orbital wall with beginning destruction of the frontal skull base was detected. Histological assessment of a biopsy, which was taken by medial orbitotomy, showed giant-cellular and fibrohistiocytic changes. Definitive histological diagnosis after removal showed an aneurysmal bone cyst. If there is evidence for aggressive, expansive growth, an aneurysmal bone cyst should be included into the ENT-differential diagnosis of orbital tumors. It is not possible to confirm diagnosis from clinical or radiological data. Early biopsy is essential for a reliable diagnosis even if histological assessment is challenging.

Stromal expression of c-Ets1 transcription factor correlates with tumor invasion.

The stroma reaction has an important role in tumor growth, invasion, and metastasis. In various invasive human carcinomas, as well as in a mouse model for tumor invasion, transcripts encoding the transcription factor c-Ets1 were detected within stromal fibroblasts, whereas they were absent in epithelial tumor cells. This expression of c-Ets1 was often increased in fibroblasts directly adjacent to neoplastic cells. Endothelial cells of stromal capillaries were also positive for c-Ets1 expression. In contrast, fibroblasts of corresponding noninvasive lesions and of normal tissues were consistently negative. In cultured human fibroblasts stimulated by basic fibroblast growth factor and tumor necrosis factor alpha, the expression of c-Ets1 correlated with the accumulation of transcripts for potential target genes, collagenase-1 and stromelysin-1. The same correlation was observed in some of the invasive carcinomas investigated. These results suggest that c-Ets1 participates in the regulation of tumor invasion in vivo.

Expression of the Ets-1 transcription factor in human astrocytomas is associated with Fms-like tyrosine kinase-1 (Flt-1)/vascular endothelial growth factor receptor-1 synthesis and neoangiogenesis.

Marked neovascularization and vascular endothelial proliferation are characteristic features of malignant gliomas. Vascular endothelial growth factor (VEGF), an angiogenic protein secreted by glioma cells, appears to play a crucial role for induction of neoangiogenesis. The VEGF receptors fms-like tyrosine kinase-1 (Flt-1)/VEGFR-1 and kinase insert domain-containing receptor (KDR)/ VEGFR-2 are up-regulated on the surface of endothelial cells (ECs) in gliomas. Both receptor genes contain an Ets-responsible element in their promoters. The proto-oncogene ets-1 encodes a transcription factor that has been associated with blood vessel formation in vivo under physiological and pathophysiological conditions including tumor neovascularization. Ets-1 is induced by VEGF in cultured ECs. In vitro data also point to a role of Ets-1 as a transcriptional activator of Flt-1. These properties prompted us to investigate Ets-1 expression in 32 human astroglial tumors of WHO grades I-IV and to correlate the data with the expression pattern of VEGF, Flt-1, and KDR. By in situ hybridization, high ets-1 mRNA levels were found in the glioma microvasculature with particularly prominent signals in glomeruloid vascular endothelial proliferations of glioblastomas (WHO grade IV). Semiquantitative reverse transcription-PCR identified the full-length ets-1 transcript but none of three known splice variants encoding isoforms with different functional domains. Immunohistochemical staining demonstrated Ets-1 protein preferentially in the nucleus of those ECs with an epithelioid morphology consistent with an activated state, whereas quiescent flat-shaped ECs predominantly displayed cytosolic immunoreactivity. This observation proposes nuclear translocation of Ets-1 during neoangiogenesis. VEGF synthesis by glioma cells was accompanied by Ets-1 expression in adjacent microvascular ECs. Furthermore, a highly significant correlation was observed between Ets-1 and Flt-1 (but not KDR) expression in ECs of the glioma microvasculature. Our data suggest that VEGF secreted by glioma cells induces Ets-1 in adjacent microvascular ECs, which subsequently transactivates the VEGF receptor Flt-1. This cascade may crucially promote neoangiogenesis in human gliomas.

Presence of genetic alterations in microdissected stroma of human colon and breast cancers.

We show by laser-assisted microdissection that frequent genetic alterations in non-hereditary invasive human colon and breast cancers (loss of heterozygosity and TP53 mutations) occur not only in the neoplastic epithelial cells, but also in the adjacent fibroblastic tumor stroma and that both components can share clonal features. Tumor cell-mesenchyme transitions are among the possible explanations for these findings.

[Lymph vascularity and lymph node metastases on PET and PET-CT: immunohistological and clinical observations]. / Das Lymphgefässsystem (LGS I und II) aus chirurgischer Sicht. Lymphgefässe und Lymphknotenabsiedlungen, nach PET, PET-CT, immunhistologischen und klinischen Beobachtungen.

Sarcomatous malignancies only rarely develop regional lymph node metastases: about 2.7% of our evaluated cases. In this paper we provide evidence supporting a new hypothesis that two entirely separate lymph vascular systems exist in humans. One system (LGS I) exists in close proximity to the epithelium and drains into regional lymph nodes. Only sarcomas that originate in the epithelium or its immediate proximity are able to form regional lymph node metastases. The vast majority of sarcomatous malignancies (97.4% of cases) do not give rise to lymph node metastases, since they originate in proximity to a second, more deeply localized lymph node system (LGS II) in the mesenchymally derived tissues of the body. This second system has no connection to regional lymph nodes. Supporting evidence is provided by experience in the operative treatment of extremity lymphedema, PET-CT examinations, radionuclear lymphography, and scientific investigations using antibodies specifically directed at the elements of the lymph vascular system.

Neointimal thickening after balloon denudation is enhanced by aldosterone and inhibited by spironolactone, and aldosterone antagonist.

OBJECTIVE: The aim was to examine the effects of aldosterone and of an aldosterone antagonist, spironolactone, on neointimal thickening in a rabbit model of balloon injury. METHODS: Eighteen rabbits underwent aortic and iliac balloon injury and were randomised to subcutaneous infusion of aldosterone (70 micrograms.kg-1.d-1) or vehicle solution for 28 d. Eighteen other rabbits were randomised to receive daily subcutaneous injections of spironolactone (50 mg.kg-1.d-1) or of vehicle for 7 d before injury and for 28 d after the procedure. All animals were then killed just after measurement of plasma renin activity and of arterial pressure. Vessels were fixed and five cross sections were analysed per rabbit (three aortic; two from iliac artery). Mean values of neointimal area and of the neointimal area/medial area ratio were calculated. RESULTS: Aldosterone treatment was associated with a decrease in renin activity and a non-significant increase in mean arterial pressure. Aldosterone significantly augmented the neointimal thickening in the iliac artery [0.42(SEM 0.07) v 0.24(0.03) mm2, P < 0.05] but not in the aorta [0.63(0.08) v 0.59(0.12) mm2, NS]. Spironolactone significantly inhibited intimal thickening, both in the iliac artery [0.09(0.02) v 0.29(0.01) mm2, P < 0.001] and in the aorta [0.31(0.03) v 0.59(0.06) mm2, P < 0.001]. Spironolactone administration was associated with an increase in renin activity and a decrease in mean arterial blood pressure. CONCLUSIONS: Aldosterone administration enhances neointimal thickening after injury and spironolactone, an aldosterone antagonist, is a potent inhibitor of neointimal thickening in the same model. This suggests a role for aldosterone in the pathophysiology of neointimal proliferation after balloon injury and for aldosterone antagonists in its prevention.

Heparin does not inhibit oncogene induction in rabbit aorta following balloon denudation.

OBJECTIVE: Smooth muscle cell proliferation and migration are the predominant responses to intimal and medial injury after percutaneous transluminal coronary angioplasty. The in vivo inhibitory effect of heparin on these responses is well documented. To test the hypothesis that the antiproliferative effect of heparin in vivo may be related to an inhibition of proto-oncogene expression, the effects of pretreatment with heparin on the expression of the c-myc, c-fos and c-jun proto-oncogenes were examined in a rabbit model of balloon denudation. METHODS: Animals were randomised 5 h before balloon denudation to receive a subcutaneous injection of unfractionated heparin (7500 IU.kg-1, n = 7) or saline (n = 6). Total RNA extracted from the aorta 1 h after balloon denudation was analysed by northern blot technique. A histological study was also performed in saline treated (n = 4) and heparin treated (n = 4) animals 28 d after balloon denudation. RESULTS: The histological study showed that the degree of neointimal thickening was significantly less in heparin treated animals. However, the level of expression of the proto-oncogenes we studied was similar in both groups. CONCLUSIONS: Heparin inhibits neointimal thickening after balloon denudation. This inhibition is not associated with an overall decrease in the level of expression of the c-myc, c-fos, or c-jun proto-oncogenes in the arterial wall, suggesting that the antiproliferative effect of heparin may be due to an effect on other events in the cell cycle.

Angiotensin converting enzyme inhibition prevents proto-oncogene expression in the vascular wall after injury.

OBJECTIVES: Angiotensin converting enzyme (ACE) inhibitors reduce neointimal hyperplasia after balloon denudation, but the mechanisms are not completely understood. It has been demonstrated that nuclear oncogenes are induced in the vascular wall in the hours immediately after injury, and that the same genes are induced by angiotensin II in vascular smooth muscle cells. It has therefore been suggested that the effects of ACE inhibitors on the response of the vessel wall could be mediated by an inhibition of proto-oncogene expression. METHODS AND RESULTS: Sixteen New Zealand White rabbits were randomly assigned for histologic analysis to receive placebo (n = 9) or 1 mg/kg per day perindopril (n = 7). After treatment for 7 days balloon aortic injury was performed. The treatment was continued and the rabbits were killed 28 days after injury. In the perindopril group the neointimal cross-sectional area was significantly smaller than in the control group. Six untreated rabbits were used to assess the time course of proto-oncogene expression in the aortic wall after injury in the present model. After extraction, total aortic RNA was hybridized with myc, fos and jun probes. Based on the results, the effects of ACE inhibition on proto-oncogene expression were tested 1 h after balloon denudation. Accordingly, 24 rabbits were randomly assigned to pretreatment for 7 days with placebo or with 1 or 10 mg/kg per day perindopril (n = 8, for each group) and were killed 1 h after injury. Expression of c-myc was not altered by pretreatment. However, 1 mg/kg per day perindopril induced significant reductions of 50% in c-jun and 45% in c-fos expression compared with control. No additional effect was obtained with the higher dose. CONCLUSION: The effect of ACE inhibition on intimal hyperplasia is associated with a reduction in early cellular events such as c-fos and c-jun expression. These results suggest that potent ACE inhibition at the time of vascular injury may be required to limit the hyperplastic response of the vessel wall.

Distribution of basement membranes in primary and metastatic carcinomas of the prostate.

The presence of periacinar and pericellular basement membranes (BMs) has been reported recently in common prostatic adenocarcinomas. In this study we extended our investigations of BMs on lymph node and hematogenous metastases, primary prostatic cancer with unusual histologic features, and posttreatment tumors. In contrast to prostatic malignancies that derive from the transitional epithelium (squamous cell carcinoma, prostatic transitional cell carcinoma) and prostatic involvement by bladder cancer, inconspicuous stromal changes and distinct BM formations at the site of tumor invasion were observed in carcinomas deriving from the secretory epithelium (papillary ductal carcinoma) and from the basal cell (basal cell carcinoma). Even highly malignant anaplastic and small cell carcinomas, as well as irradiated and/or hormonally treated tumors, showed distinct BM formations in contact with the stroma. The same observations could be made in lymphatic and hematogenous metastases of different anatomic sites. These findings indicate that prostatic malignancies may retain BMs even in high-grade lesions, metastases, posttreatment tumors, and variants of prostatic adenocarcinoma.

The multiple roles of tumour stroma.

Since the work of Judah Folkman in the 1970s demonstrating the importance of vascularization on tumour growth the many roles played by tumour stroma have been demonstrated. Vascular endothelial growth factor/vascular permeability factor appears to be the main in vivo inducer of both stromal blood vessels and other components of the tumour stroma. Its action is probably mediated through its strong and long-lasting effect on microvascular permeability leading to fibrin extravasation and organisation ("tumours are wounds that do not heal"). During tumour invasion, stromal fibroblasts participate in the degradation of the extracellular matrix (ECM) by secreting matrix degrading proteases as well as their downstream-activators. Stroma derived factors such as scatter factor/hepatocyte growth factor as well as interactions between neoplastic cells and the ECM can play a role in both tumour cell migration and proliferation. The ECM may also act as a reservoir for growth factors. A novel transcription factor encoded by the c-ets 1 proto-oncogene is likely to be involved in the transcriptional regulation of both tumour vascularization and invasion. This contribution summarizes recent developments in the tumour stroma field.

Invasive properties of serous human epithelial ovarian tumors are related to Ets-1, MMP-1 and MMP-9 expression.

The invasive potential of serous epithelial ovarian tumors is the main factor determining their biological behaviour. In contrast to invasive serous ovarian carcinomas serous borderline tumors generally present without stromal invasion and without or non-invading peritoneal implants. Little is known about the reasons underlying these differences. In the present study we found that two matrix-degrading metalloproteinases, collagenases 1 and 4 (MMPs 1 and 9), as well as the Ets-1 transcription factor are expressed at very low levels in serous benign cystadenomas, upregulated in the fibroblastic stroma, but not in the epithelium of borderline tumors and most strongly expressed in both stromal and epithelial tumor cells of serous invasive carcinomas. Since expression of Ets-1 and of MMPs 1 and 9 are topographically related, a transcriptional regulation of both proteases by Ets-1 is suggested. Upregulation of MMPs 1 and 9 within fibroblastic stromal cells of borderline tumors might be related to matrix remodelling and additional expression of both enzymes by the neoplastic cells of invasive carcinomas could then allow invasive propagation. The different expression patterns might supports the view, that no transition of serous borderline tumors into invasive carcinomas occurs.

Pathological aspects of prostate cancer and benign nodular hyperplasia.

The aim of the study was to investigate tumor suppressor genes (TSGs) which play a role in the pathogenesis of prostate cancer. Additionally, different prostate tumors were immunohistochemically related to their potential precursor cells, the basal cells and the glandular secretory epithelium, which differ in their hormone responsiveness. By PCR-amplification of microsatellite-DNA we found allelic losses of chromosomal loci near known or putative TSGs to increase with the malignancy grade of prostate carcinoma. When investigated for numerous markers common and endometrioid carcinoma were immunohistochemically related to the secretory epithelium while the rare basal cell tumor, containing the estrogen receptor, squamous cell carcinoma and urothelial carcinoma showed features of the basal cells. In histopathological differential diagnosis high molecular weight basal cell keratins, prostatic acid phosphatase and prostate specific antigen may be of value. Stromal type nodular hyperplasia and the fetal prostate mesenchyme had common immunohistochemical features which might reflect analogous development.

Presence of genetic alterations in microdissected stroma of human colon and breast cancers.

BACKGROUND: Human carcinomas not only consist of neoplastic epithelial cells but also of tumor stroma, which may play an important role in tumor-progression. Whereas the tumor surrounding stroma is generally believed to represent a reactive component induced by tumor cell derived factors, a contribution of neoplastic cells to stroma formation via epithelium-mesenchyme transition during tumor invasion has become a novel concept in recent years. MATERIALS, METHODS AND RESULTS: We here show, by laser-assisted microdissection, that frequent genetic alterations in non-hereditary invasive human colon and breast cancers (loss of heterozygosity and TP53 mutations) occur not only in the neoplastic epithelial cells, but also in the adjacent fibroblastic stroma and that both components can share clonal features. CONCLUSION: Tumor cell-mesenchyme transitions are among the possible explanations for these findings and could actually occur during tumor invasion in vivo.

Implication of the proliferation and apoptosis associated CSE1L/CAS gene for breast cancer development.

The CSEIL/CAS protein (CAS) is a Ran-binding protein with a function as a nuclear transport (export) factor. CSEIL/CAS, similar to Ran and other ran-binding proteins, plays at the same time an important role in the mitotic spindle checkpoint, which assures genomic stability during cell division. This checkpoint is frequently disturbed in neoplasms of various origin, including breast, hepatic and colonic tumors. CAS is located on chromosome 20ql3 and amplified in several cell lines, including breast, colon and bladder cancer. MEKl phosphorylation is known to be a reason for different CAS localization and activity. We evaluated the expression of CAS in 50 benign and malignant tumors of the breast by immunohistochemistry. Benign lesions of the breast (n=13) revealed a weak, predominantly cytoplasmatic CAS positivity. In ductal and lobular in situ carcinomas (n=17), 70-90% of the tumor cells were positive for anti-CAS staining which was predominantly cytoplasmatic. In invasive ductal and lobular carcinomas (n =20), 70-90% of the tumor cells stained positive with anti-CAS in a predominantly nuclear pattern. Different localization of CAS might affect its role not only for chromosome segregation, proliferation and apoptosis, but also its function in nuclear transport of proteins like retinoblastoma-gene-product, p53 and BRCAl. A different regulation in this checkpoint might contribute to the invasive potential in malignant carcinomas of the breast. Alteration of CAS-activity, possibly via MEKl-inhibition, might therefore be a possible option for breast cancer therapy.

Histology and immunohistochemistry studies in prostate cancer.

Histopathologic diagnosis of prostate carcinoma is not yet free of problems. There are particular difficulties in demarcating atypical forms of hyperplasia from well-differentiated carcinomas and in diagnosing and classifying incidental carcinomas. With the aid of conventional histology, however, nearly all diagnostic problems relevant for the patient can be solved. In routine diagnostics, modern immunohistochemical techniques are particularly helpful in assessing metastases of an undetected primary tumor. New immunohistochemical techniques provide insight into the receptor content of the prostate and show the histogenesis of prostate carcinoma in a new light. The estrogen receptor (modified ER-ICA test) is present in the nuclei of stromal cells and of basal cells within the glands, but not in the secretory epithelium. The receptor-associated protein--ER-D5--is found in the cytoplasm of stromal and basal cells. In basal cells and secretory epithelium, keratins show a different pattern. Immunohistochemically common adenocarcinomas display the pattern of secretory epithelium; urothelial and squamous cell carcinomas, on the contrary, display the pattern of basal cells. This finding does not support the opinion that the basal cell is the stem cell of secretory epithelium and the precursor cell of prostate carcinoma.

Excessive epithelioid cell granulomatous reaction associated with a lymphoepithelial carcinoma (Schmincke-Regaud).

We report on a 56-year-old man in whom histologic examination of "granulation tissue" from the right lower meatus of the nose revealed the presence of an undifferentiated nasopharyngeal carcinoma of the lymphoepithelial type (Schmincke-Regaud). The diagnosis was supported by serologic detection of antibodies against the Epstein-Barr virus. Approximately 3 weeks after the first excision, biopsy material was taken from the epipharynx. Histologically a marked epithelioid cell granulomatous reaction was found. As an unusual feature, granulomas were not located between tumor cell complexes, but rather contained tumor remnants in their center. At tissue site all transitions from well preserved to completely necrotic tumor cells were discernible. This unusual reaction probably represents a strong, favorable immunologic response against the neoplasm.

Immunohistochemical estrogen receptor demonstration in the prostate and prostate cancer.

Immunohistochemical investigations of human prostate and human prostate cancer were performed using two different monoclonal antibodies for the demonstration of estrogen receptors (ER). One marked the nuclear estrogen receptor protein (ERICAR, Abbott Laboratories), whereas the other one marked an estrogen receptor-associated cytoplasmic protein (ER-D5 kit, Amersham Buchler). 12 transurethral resection specimens with benign prostatic hyperplasia and 10 prostatic carcinomas were investigated by ERICA. Only in 1 specimen was a focally ERICA-positive basal cell hyperplasia found. The ER-D5 kit yielded a constantly positive reaction of stromal cells, especially smooth muscle cells. Basal cell hyperplasia and squamous metaplasia, alterations which can occur during estrogen application, were always strongly ER-D5-positive. Eleven (13.4%) out of 82 prostate carcinomas were focally positive. The staining results with ER-D5 are in good accordance with biochemical estrogen receptor investigations which demonstrated the estrogen receptors especially in the fibromuscular stroma. Furthermore, the results show that therapeutically applied estrogens do not directly inhibit the growth of prostatic carcinoma, however, the effect is an indirect one by suppressing androgen synthesis of the testis. The different staining results with ER-D5 and ERICA can be explained by the low ER concentration of prostate, the concentration is evidently below the limits of detectability with ERICA.