Erich-Franz Solomayer

Bisphosphonate-induced osteonecrosis of the jaw (ONJ): Incidence and risk factors in patients with breast cancer and gynecological malignancies

OBJECTIVEnSince 2003, multiple cases of bisphosphonate-induced osteonecrosis of the jaw (ONJ) were reported. The aim of this study was to describe the incidence and risk factors of ONJ in patients with breast cancer or gynecological malignancies receiving bisphosphonates (BP).nnnMETHODSnONJ was recorded for all patients with breast cancer or gynecological malignancies treated with intravenous bisphosphonates at the Department of Gynecology and Obstetrics, University Hospital Tuebingen during April, 1999 and May, 2006.nnnRESULTSn10 of 345 (2.9%) patients with breast cancer or gynecological malignancies developed ONJ while receiving bisphosphonate therapy. Six patients with ONJ had a history of recent dental procedures. All patients had received zoledronic acid as part of their bisphosphonate regimen. Time of exposure to bisphosphonates and the number of treatment cycles were significant risk factors for the development of ONJ (p<0.001). In patients diagnosed with ONJ the mean number of treatment cycles was 27+/-18 cycles. However, the mean number of treatment cycles in patients without manifestation of ONJ was 12+/-12 cycles.nnnCONCLUSIONnLength of exposure to BPs and the cumulative dose of given BPs seem to be the most important risk factors for the development of ONJ followed by dental procedures.

ERalpha-status of disseminated tumour cells in bone marrow of primary breast cancer patients

IntroductionIsolated disseminated tumour cells (DTC) are regarded as surrogate markers for minimal residual disease in breast cancer. Characterisation of these cells could help understand the known limitations of adjuvant therapy. Of particular interest is their oestrogen-receptor (ER) status because endocrine adjuvant therapy remains a cornerstone of breast cancer treatment.MethodsBone marrow (BM) aspirates from 254 patients with primary breast cancer were included in this study. A double immunofluorescence staining procedure was established for the identification of cytokeratin (CK) positive/Erα-positive cells. ERα status of the primary tumour was assessed immunohistochemically using the same antibody against ERα.ResultsIn 107 of 254 (42%) breast cancer patients, CK-positive cells could be detected in the BM. More than one DTC in the BM was observed in 38 of the 107 patients. The number of detected cells ranged between 1 and 55 cells per 2 × 106 mononuclear cells. DTCs demonstrated ERα positivity in 12% of the patients. The ERα expression was heterogeneous in 10 of the 38 (26%) patients with more than one DTC. The concordance rate of ERα status between primary tumour and DTC was 28%. Only 12 of 88 patients with ERα-positive tumours also had ERα-positive DTCs.ConclusionsPrimary tumours and DTCs displayed a concordant ERα status in only 28% of cases. Most of the DTCs were ERα negative despite the presence of an ERα-positive primary tumour. These findings further underline the distinct nature of DTCs and may explain the failure rates seen in conventional endocrine adjuvant therapy.

Presence of apoptotic and nonapoptotic disseminated tumor cells reflects the response to neoadjuvant systemic therapy in breast cancer

IntroductionNeoadjuvant systemic therapy (NST) is an established strategy to reduce tumor size in breast cancer patients prior to breast-conserving therapy. The effect of NST on tumor cell dissemination in these patients is not known. The aim of this study was to investigate the incidence of disseminated tumor cells (DTC), including apoptotic DTC, in breast cancer patients after NST, and to investigate the correlation of DTC status with therapy response.MethodsBone marrow aspiration was performed in 157 patients after NST. DTC were detected by immunocytochemistry using the A45–B/B3 anticytokeratin antibody. To detect apoptotic DTC the antibody M30 (Roche Diagnostics, Germany) was used, which detects a neo-epitope expressed only after caspase cleavage of cytokeratin 18 during early apoptosis.ResultsThe incidence of DTC in breast cancer patients was 53% after completion of NST. Tumor dissemination was observed more frequently in patients with no change/progressive disease (69%) than in patients with partial remission or complete remission of the primary tumor (46%) (P < 0.05). Ten out of 24 patients with complete remission, however, were still bone marrow positive. Apoptotic DTC were present in 36 of 157 (23%) breast cancer patients. Apoptotic cells only were detected in 14% of the patients with partial remission or complete remission, but were detected in just 5% of the patients with stable disease. Apoptotic DTC were detectable in none of the patients with tumor progression.ConclusionThe pathological therapy response in breast cancer patients is reflected by the presence of apoptotic DTC. Patients with complete remission, however, may still have nonapoptotic DTC. These patients may also benefit from secondary adjuvant therapy.

Membrane-initiated effects of progesterone on proliferation and activation of VEGF in breast cancer cells

Objective Progesterone influences mammary gland development and probably breast cancer tumorigenesis and functions by regulating a broad spectrum of physiological processes. We investigated receptor membrane-initiated actions of progesterone in MCF-7 breast cancer cells via progesterone receptor membrane component 1 (PGRMC1). Design and method The expression of PGRMC1 in breast cancer was verified by immune fluorescent analysis of paraffin sections. MCF-7 cells were transfected with PGRMC1 (wild type) or PGRMC1 variants. These cells were stimulated with a membrane-impermeable progesterone (P4) conjugate (P4-BSA-fluorescein isothiocyanate, P4-BSA-FITC, 10−6 mol/l) or unconjugated progesterone (P4, 10−6 mol/l) in the presence or absence of the progesterone receptor blocker RU-486 (10−6 mol/l). Additionally, the effects on the expression of vascular endothelial growth factor A (VEGF-A) were determined using quantitative real-time polymerase chain reaction. Results PGRMC1 is perinuclearly localized in breast cancer cells. Western Blot analysis suggests that PGRMC1 is phosphorylated at serine 180. MCF-7-PGRMC1 (S180A) cells show an approximately 35% increase in proliferation after incubation with P4-BSA-FITC compared to MCF-7 control and MCF-7-PGRMC1 (wild type) cells. This effect cannot be blocked by RU-486. P4 reduced proliferation of MCF-7-PGRMC1 cells by approximately 10% compared to untreated controls. P4-BSA-FITC treatment led to a roughly three-fold activation of VEGF-A gene expression compared to MCF-7 cells. Conclusion PGRMC1 is expressed in breast cancer tissue and mediates an RU-486-independent proliferative signal. It might also contribute to VEGF-induced neovascularization in tumor tissue. Thus, screening for PGRMC1 expression might be of interest to identify women with a higher expression of PGRMC1 and who might thus be susceptible for breast cancer development under hormone replacement therapy. H. Neubauer and G. Adam contributed equally to this manuscript.

Leiomyoma in the Retzius space: a rare cause for voiding difficulties

A 54-year-old woman with voiding difficulties was referred to our department. She complained about a slow urine stream, hesitancy, straining when voiding and the feeling of incomplete emptying. The gynaecological examination revealed a 4u2009×u20093xa0cm pelvic tumour. The tumour was well circumscribed in the retropubic space between the symphysis and the bladder neck sonographically and by magnetic resonance imaging and was closed off from neighbouring structures. After removal of the tumour, the voiding problems were reversed, and the patient has remained asymptomatic. The histological examination showed a leiomyoma with high vascularisation. This case report showed that retropubic tumours can obstruct the urethra and cause voiding dysfunctions. Consequently, this needs to be considered in the differential diagnosis.

Abdominal Hysterectomy: Indications and Contraindications

Hysterectomy is the most common non-obstetric gynecologic procedure and remains the gold standard for treatment of benign uterine pathologies and part of the surgical treatment of gynecologic malignancies. Many guidelines have been published regarding the best technique to perform hysterectomy and although overall recommendations endorse a vaginal or minimal invasive approach whenever feasible, the largest proportions of hysterectomies worldwide are performed via laparotomy. This chapter gives an overview of the indications and contraindications of abdominal hysterectomy and introduces factors which influence the choice of the route of hysterectomy.

Comparison between circulating and disseminated tumor cells in breast cancer

Background: The presence of disseminated tumor cells in bone marrow (BM) of breast cancer patients is an independent prognostic factor. The role of circulating tumor cells (CTC) in blood is not yet defined. Since BM aspiration is less accepted by patients compared to blood drawing, it would be highly desirable to replace bone marrow aspiration by blood analysis. Therefore, the purpose of the present study was: to examine the presence of tumor cells in peripheral blood, to evaluate how surgery affects the presence of CTC, to assess the correlation between results in blood and in BM.

Expression and functional analysis of PGRMC1 in breast cancer

5178 Background:u2028 In a proteomic screening approach we identified progesterone receptor membrane component 1 (PGRMC1) as a protein upregulated in frozen ER-negative breast cancer tissue samples making PGRMC1 a potential therapeutic target. PGRMC1 is a 28kDa protein consisting of 194 amino acids and belongs to the “membrane-associated progesterone receptor” (MAPR) protein family.u2028 Aim:u2028 The aim of this project is to characterize the potential biological function of PGRMC1 and its expression in breast cancer.u2028 Material and Methods:u2028 Structure analysis was performed (http://scansite.mit.edu/motifscan_id.phtml) under high and medium stringency to identify protein motifs predicted for PGRMC1. Based on these predicted motifs PGRMC1-variants were generated by site-directed mutagenesis and MCF-7 breast cancer cells were stably transfected with the corresponding expression plasmids. Several PGRMC1-specific hybridoma were generated and tested by western blot analysis. Multiple immune fluorescence for PGRMC1, estrogen receptor alpha and the hypoxia marker glucose transporter 1 (Glut1) was applied to label tissue microarrays containing different breast cancer specimen. Additionally, posttranslational modification of PGRMC1 was investigated by phosphatase treatment of tissue lysates. Functional analysis was performed by stimulating transfected MCF-7 cells with a membrane-impermeable pro-gesterone:BSA:fluoresceinisothiocyanate conjugate followed by analysis of cell proliferation by measuring the cellular ATP content.u2028 Results:u2028 PGRMC1 contains a cytochrome b5 domain and several protein interaction domains making it a possible signalling adaptor protein. Further, PGRMC1 is posttranslationally modified. Stimulation of PGRMC1 expressing MCF-7 cells resulted in an increased proliferation compared to control cells grown in growth factor and hormone reduced medium. Western blot analysis of hybridoma produced a PGRMC1-specific signal at the predicted molecular weight of 28 kDa which is not detected after preincubation of the hybridoma with recombinant PGRMC1 protein. In immune fluorescence analysis of breast cancer tissue sections presumably myoepithelial cells were highly positive for PGRMC1. Besides that PGRMC1 expression was upregulated in Glut1 positive, hypoxic areas in ductal carcinoma in situ of comedo type.u2028 Conclusion:u2028 These data indicate that PGRMC1 is expressed in breast cancer and might functionally play a role in cell proliferation. Further determination of the so far poorly defined role of PGRMC1 in cancer biology could prove to be of great relevance to clinical cancer therapists.