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Small cell lung cancer (SCLC) represents 15% of lung cancers and is characterized by early dissemination, development of chemoresistance and a poor prognosis. A host of diverse drugs failed invariably and its mechanisms of global chemoresistance have not been characterized so far. SCLC represents the prototype of an aggressive and highly metastatic tumor which is ultimately refractory to any treatment. High numbers of circulating tumor cells (CTCs) allowed us to establish 5 CTC cell lines (BHGc7, 10, 16, 26 and UHGc5) from patients with recurrent SCLC. These cell lines exhibit the typical SCLC markers and CTCs of all patients developed spontaneously large multicellular aggregates, termed tumorospheres. Ki67 and carbonic anhydrase 9 (CAIX) staining of tumorosphere sections revealed quiescent and hypoxic cells, respectively. Accordingly, comparison of the chemosensitivity of CTC single cell suspensions with tumorospheres demonstrated increased resistance of the clusters against chemotherapeutics commonly used for treatment of SCLC. Therefore, global chemoresistance of relapsing SCLC seems to rely on formation of large tumorospheres which reveal limited accessibility, lower growth fraction and hypoxic conditions. Since similar tumor spheroids were found in other tumor types, SCLC seems to represent a unique tumor model to study the association of CTCs, metastasis and drug resistance.

Introduction: Brigatinib is a second-line inhibitor for the treatment of rearranged anaplastic lymphoma kinase (ALK) in lung cancer patients which has significant activity against brain metastases. This tyrosine kinase inhibitor (TKI) overcomes a wide range of ALK mutations which confer therapeutic resistance and is increasingly applied in first-line therapy due to improved benefit for patients compared to crizotinib, the current standard of care. Areas covered: The authors review the development and characteristics of brigatinib and discuss the optimal clinical use and sequence of the application of ALK inhibitors in patients progressing under therapy. Expert opinion: ALK-rearranged NSCLC can be treated with a broad range of approved and novel inhibitors at various stages of therapy, including the second-line therapeutic brigatinib. Besides this TKI, the second-line ALK inhibitors alectinib and ceritinib, as well as the third-line lorlatinib are approved for the treatment of ALK-positive NSCLC patients. The main challenge is to find sequences and combinations of ALK inhibitors which provide the best benefit for the patients.

Small cell lung cancer (SCLC) is disseminated in the majority of patients at first presentation and, thus, treated with chemoradiotherapy. Despite initial high response rates, chemoresistance appears rapidly and results in a dismal prognosis. However, patients with limited cancer may exhibit better disease control upon surgical treatment. Correct staging is highly critical in the selection of those patients which are likely to benefit from surgery. Studies of the inclusion of surgery in the multimodal treatment of SCLC vary widely in number of patients, selection, treatment and follow-up. Nevertheless surgical therapy for confined SCLCs achieves favorable long-term survival compared to chemoradiotherapy, depending on a precise assessment of the degree of tumor dissemination. Recently, extremely high counts of circulating tumor cells (CTCs) were reported in patients with SCLC compared to other malignancies. In several studies the number of CTCs was found to constitute a prognostic parameter and a marker of response to therapy. Therefore, the assessment of CTCs as so-called "Liquid Biopsy" seems to constitute a more precise method to detect tumor dissemination earlier when compared to clinical staging. In conclusion, in the era of precision oncology enumeration and identification of CTCs of SCLC patients have the potential to help in the selection of patients most suitable for tumor surgery.

Historically disseminated tumor cells in solid cancers were detected by immunohistochemistry of bone marrow aspirates. Blood samples are more easily and frequently accessible and are therefore employed to detect circulating tumor cells (CTCs), a procedure termed “liquid biopsy”. CTCs are shed from tumors and a fraction of these cells is capable of establishing distal lesions. Since patients die from metastatic disease, CTCs hold potential to have prognostic significance and to indicate the success or failure of therapeutic interventions. With few exceptions, CTCs are rare cells against a large background of blood cells and need to be enriched for detection and characterization. The FDA-approved CellSearch© system relies on the selection of epithelial cell adhesion molecule (EpCAM)-positive tumor cells and is incorporated in numerous clinical studies for risk stratification and assessment of therapy. However, CTCs may have undergone (partial) epithelial-mesenchymal transition and are therefore missed by methods dependent on epithelial markers. A host of other techniques to enrich/isolate CTCs which are based on physical and/or distinct biological properties are under evaluation. Although single CTCs were linked to an increased risk of tumor progression, the actual phenotype and fraction of these cells capable of forming metastases are not known, as well as the mechanisms and kinetics of tumor cell shedding and evasion. Despite considerable progress, CTC detection methods still need to be validated and their clinical significance confirmed in larger multicenter trials before entering clinical routine practice.

Small cell lung cancer (SCLC) accounts for 15 % of all lung tumors and represents an invasive neuroendocrine malignancy with poor survival rates. This cancer is highly prevalent in smokers and characterized by inactivation of p53 and retinoblastoma. First in vitro expansion of circulating tumor cells (CTCs) of SCLC patients allowed for investigation of the cell biology of tumor dissemination. In the suggested CTC SCLC model, the primary tumor attracts and educates tumor-promoting and immunosuppressive macrophages which in turn arm CTCs to spread and generate distal lesions. Preexisting inflammatory processes associated with chronic obstructive pulmonary disease (COPD) seem to potentiate the subsequent activity of tumor-associated macrophages (TAM). Activation of signal transducer and activator of transcription 3 (STAT3) and expression of chitinase-3-like 1/YKL-40 in SCLC CTCs seems to be associated with drug resistance. In conclusion, inflammation-associated generation of invasive and chemoresistant CTCs most likely explains the characteristic features of SCLC, namely early dissemination and rapid failure of chemotherapy.

Small cell lung cancer (SCLC) has a poor prognosis and is found disseminated at first presentation in the majority of cases. The cell biological mechanisms underlying metastasis and drug resistance are not clear. SCLC is characterized by high numbers of circulating tumor cells (CTCs) and we were able to expand several CTC lines ex vivo and to relate chitinase-3-like-1/YKL-40 (CHI3L1) as marker. Availability of expanded SCLC CTC cells allowed for a screening of receptor tyrosine kinases (RTKs) expressed. The metastatic CHI3L1-negative SCLC cell line SCLC26A, established from a pleural effusion was used for comparison. The CTC cell line BHGc10 was found to exhibit increased expression of RYK, AXL, Tie-1, Dtk, ROR1/2, several ephrins (Eph) and FGF/EGF receptors compared to SCLC26A. All of these RTKs have been associated with cell motility, invasion and poor prognosis in diverse cancer entities without knowledge of their association with CTCs. The identification of RYK, AXL and ROR1/2 as pseudokinases, lacking activity, seems to be related to the observed failure of RTK inhibitors in SCLC. These kinases are involved in the noncanonical WNT pathway and their expression in SCLC CTCs represents a cancer stem cell-like phenotype.

Cancer cell lines are good in vitro models to study molecular mechanisms underlying chemoresistance and cancer recurrence. Recent works have demonstrated that most of the available ovarian cancer cell lines are most unlikely high grade serous (HGSOC), the major type of epithelial ovarian cancer. We aimed at establishing well characterized HGSOC cell lines, which can be used as optimal models for ovarian cancer research. We successfully established seven cell lines from HGSOC and provided the major genomic alterations and the transcriptomic landscapes of them. They exhibited different gene expression patterns in the key pathways involved in cancer resistance. Each cell line harbored a unique TP53 mutation as their corresponding tumors and expressed cytokeratins 8/18/19 and EpCAM. Two matched lines were established from the same patient, one at diagnosis and being sensitive to carboplatin and the other during chemotherapy and being resistant. Two cell lines presented respective BRCA1 and BRCA2 mutations. To conclude, we have established seven cell lines and well characterized them at genomic and transcriptomic levels. They are optimal models to investigate the molecular mechanisms underlying the progression, chemo resistance and recurrence of HGSOC. publisher: Elsevier articletitle: Molecular characterization of 7 new established cell lines from high grade serous ovarian cancer journaltitle: Cancer Letters articlelink: http://dx.doi.org/10.1016/j.canlet.2015.03.040 content_type: article copyright: Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd. ispartof: Cancer Letters vol:362 issue:2 pages:218-28 ispartof: location:Ireland status: published

Picoplatin was developed as platinum coordination complex to overcome development of resistance, through conjugation to thioles, by the introduction of a methyl-pyridine moiety into the cisplatin parent structure. Pharmacokinetic parameters of the drug, after intravenous and oral application, were studied in solid tumors and clinical Phase I - III trials performed, in particular in NSCLC and small cell lung cancer (SCLC). Results showed low clinical activity of picoplatin.This article presents an overview of the pharmacokinetic assessments of picoplatin in lung cancer. Specifically, the authors address the relationship between disposition and clinical activity of the drug.Picoplatin failed to overcome resistance to platinum compounds in lung cancer to achieve significant improved survival of most patients. Even highest doses of the drug reaching 150 m/m² given intravenously every 3 weeks were not sufficient to achieve better response than existing chemotherapeutics and the oral bioavailability of a dose of 200 - 400 mg corresponded only to 80 mg/m² iv. Picoplatin therefore seem to be quite ineffective. Picoplatin is expected to overcome tumor resistance in cases which overexpress thiol-conjugating pathways; however, this was not proved in clinical trials. To conclude, this blocked platinum complex is not able to reverse cisplatin resistance to a significant extent in vivo and its mechanisms and kinetics and of DNA damage failed to produce significant clinical results compared to second-line standard therapy for lung cancer.

Ulrike Olszewski, Robert Zeillinger, Klaus Geissler, Gerhard HamiltonLudwig Boltzmann Cluster of Translational Oncology, Ludwig Boltzmann Society, Vienna, AustriaPurpose: Carcinoids are highly chemoresistant tumors associated with a dismal prognosis. This study involved a comparison of the genome-wide gene expression pattern of a chemoresistant and a chemosensitive pulmonary carcinoid cell line to reveal factors that contribute to the resistant phenotype.Materials and methods: Gene expression of UMC-11 chemoresistant carcinoid cells as assessed by 32 K microarray was compared with H835 chemosensitive carcinoid cells, and the genes that were differentially expressed and expected to be related to chemoresistance were selected.Results: Drug-resistant UMC-11 cells exhibited increased expression of transcripts known to confer resistance to different cytostatics such as P-glycoprotein, multidrug resistance-associated proteins 2 and 3, effectors of the glutathione detoxification and xenobiotics degradation pathways, and ion transporters including Na+/K+-adenosine triphosphatase. In addition, enhanced transcription of several S100 proteins, capable of suppressing apoptosis, regulation of topoisomerase I (topo I) expression by antisense transcripts from TOPO1 pseudogenes, and alterations of the cytoskeleton seem to contribute to the multidrug-resistant phenotype. A multitude of epidermal growth factor (EGF)-related and neuropeptide growth factors, overexpression of proteases, and appearance of aerobic glycolytic metabolism complement the malignant phenotype of the UMC-11 cells.Conclusion: The multidrug-resistant phenotype of the UMC-11 pulmonary carcinoid cell line seems to be mediated by classical efflux pumps, drug metabolization or conjugation systems, and, possibly, modulation of apoptotic cell death by S100 proteins and topo I expression by pseudogene transcripts. Autocrine or paracrine stimulation by a host of EGF-related and neuropeptide growth factors, as well as high metastatic potency indicated by increased expression of components of aerobic glycolysis and proteolytic enzymes, may furthermore account for the failure of therapeutic interventions.Keywords: neuroendocrine tumor, drug resistance, microarray, drug transporter, apoptosis

Cancer patients die of metastatic disease but knowledge regarding individual steps of this complex process of intravasation, spread and extravasation leading to secondary lesions is incomplete. Subpopulations of tumor cells are supposed to undergo an epithelial-mesenchymal transition (EMT), to enter the bloodstream and eventually establish metastases in a reverse process termed mesenchymal-epithelial transition (MET). Small cell lung cancer (SCLC) represents a unique model to study metastatic spread due to early dissemination and relapse, as well as availability of a panel of circulating cancer cell (CTC) lines recently. Additionally, chemosensitive SCLC tumor cells switch to a completely resistant phenotype during cancer recurrence. In advanced disease, SCLC patients display extremely high blood counts of CTCs in contrast to other tumors, like breast, prostate and colon cancer. Local inflammatory conditions at the primary tumor site and recruitment of macrophages seem to increase the shedding of tumor cells into the circulation in processes which may proceed independently of EMT. Since millions of cells are released by tumors into the circulation per day, analysis of a limited number of CTCs at specific time points are difficult to be related to the development of metastatic lesions which may occur approximately one year later. We have obtained a panel of SCLC CTC cell line from patients with relapsing disease, which share characteristic markers of this malignancy and a primarily epithelial phenotype with unique formation of large tumorospheres, containing quiescent and hypoxic cells. Although smoking and inflammation promote EMT, partial expression of vimentin indicates a transitional state with partial EMT in these cell lines at most. The CTC lines exhibit high expression of EpCAM , absent phosphorylation of β-catenin and background levels of Snail. Provided that these tumor cells had ever undergone EMT, here in advanced disease MET seem to have occurred already in the peripheral circulation. Alternative explanations for the expression of mesenchymal markers of the CTC lines are the heterogeneity of SCLC cells, cooperative migration or altered gene expression in response to the inflammatory tumor microenvironment allowing for tumor spread without EMT/MET.

Objective: Vascular endothelial growth factor (VEGF) is an important angiogenic factor, and its receptors have been shown to be overexpressed in various human carcinomas. In this study, we investigated the role of scanning with iodine-123 (123I)-labelled VEGF165 in patients with highly malignant osteosarcoma. Methods: Two patients (a 15-year-old female and a 14-year-old male) with osteosarcoma were injected with 140 MBq [165 per patient] of 123I-VEGF165. Dynamic acquisition was initiated immediately after administration and carried out until 30 min after injection. Whole-body images were done in anterior and posterior views at various time points. All patients underwent single-photon emission tomography imaging. Results:123I-VEGF165 scans were positive in these patients. Sequential images clearly showed increased 123I-VEGF165 activity in osteosarcoma lesions. The tumour lesions were still visualized in whole-body images and single-photon emission tomography examinations 2 h after injection. Intravenous injection of 123I-VEGF165 did not cause any side effects. Conclusion: Our results suggest that 123I-VEGF165 receptor scintigraphy may be useful for the visualization of highly malignant osteosarcoma and/or metastasis and the angiogenic activity of the tumour.

Intermittent androgen suppression (IAS) therapy for prostate cancer patients attempts to maintain the hormone dependence of the tumor cells by cycles alternating between androgen suppression (AS) and treatment cessation till a certain prostate-specific antigen (PSA) threshold is reached. Side effects are expected to be reduced, compared to standard continuous androgen suppression (CAS) therapy. The present study examined the effect of IAS on bone metabolism by determinations of serum procollagen I N-terminal peptide (PINP), a biochemical marker of collagen synthesis. A total of 105 treatment cycles of 58 patients with prostate cancer stages ≥pT2 was studied assessing testosterone, PSA and PINP levels at monthly intervals. During phases of AS lasting for up to nine months PSA levels were reversibly reduced, indicating apoptotic regression of the prostatic tumors. Within the first cycle PINP increased at the end of the AS period and peaked in the treatment cessation phase. During the following two cycles a similar pattern was observed for PINP, except a break in collagen synthesis as indicated by low PINP levels in the first months off treatment. Therefore, measurements of the serum PINP concentration indicated increased bone matrix synthesis in response to >6 months of AS, which uninterruptedly continued into the first treatment cessation phase, with a break into each of the following two pauses. In summary, synthesis of bone matrix collagen increases while degradation decreases during off-treatment phases in patients undergoing IAS. Although a direct relationship between bone matrix turnover and risk of fractures is difficult to establish, IAS for treatment of biochemical progression of prostate tumors is expected to reduce osteoporosis in elderly men often at high risk for bone fractures representing a highly suitable patient population for this kind of therapy.

Soluble cytokeratin 18 fragments (M30, M65) are released from human cancer cells during cell death and hold potential as biomarkers in colorectal cancer characterized by frequent metastatic spread. A total of 62 colorectal cancer and 27 control patients were included in the study. M65 (necrosis and apoptosis) and M30 (apoptosis) were quantified preoperatively (n = 62) and postoperatively (n = 31) using specific enzyme-linked immunosorbent assays. Presence of disseminated tumor cells (DTC) in the bone marrow was assessed by staining of A45-B/B3-positive cells in aspirates. M65 was significantly elevated in patients with International Union against Cancer stage I and IIA tumors compared to controls. A subgroup (19/31) exhibited a significant (p0.05) decrease of M65 after tumor surgery (503.9 +/- 230.7 to 342.6 + 94.8 U/l; -32.0 +/- 16.5%), in contrast to 12 patients who revealed higher M65 levels postoperatively (386.5 +/- 128.5 to 519.1 +/- 151 U/l; +37.4 +/- 32.3%). DTC in bone marrow were found in 10% (2/19) of patients with decreasing and 50% (6/12) of the patients with increasing M65 serum concentrations after surgery (p = 0.028). In conclusion, M65 as marker is likely to be valuable to identify patients with a high incidence of systemic disease.

Lung cancer is the leading cause of cancer-related deaths worldwide, and the small cell lung cancer (SCLC) histotype is its most aggressive variant. Yu et al. recently reviewed the detection methods and clinical applications of circulating tumor cells (CTCs) in lung cancer in detail [1]. CTCs are shed from primary tumors or metastatic sites into the peripheral blood and may serve as liquid tumor biopsies for early diagnosis, risk evaluation, and monitoring of therapy [1, 2]. As CTCs are normally rare, detection methods comprise enrichment steps followed by identification with selective markers. The Cell-Search system is the only diagnostic assay approved by the US Food and Drug Administration and was employed to detect SCLC CTCs. In particular, CTCs were defined as nucleated cells coexpressing EpCAM and cytokeratins and negative for the white blood cell surface marker CD45 [2]. In SCLC patients, the detection rates of the CellSearch system ranged from 14.3 to 68.6 % [1]. However, during cancer invasion and metastasis, some cells undergo epithelial– mesenchymal transition (EMT), thus increasing the probability of false-negative CTC results. Among primary lung cancer patients, SCLC patients showed a higher CTC count than non-small cell lung cancer (NSCLC) cases [1]. A higher stage of lung cancer usually correlates with a higher number of CTCs. Yet, only a minority of these disseminated cells can develop into metastases [2]. Lung cancer cells shed into the circulation during surgery only persist for a short time [1]. Numerous investigations have evaluated the prognostic value of CTCs in lung cancer; however, the conclusions of these studies are controversial. For example, Naito et al. indicated that SCLC patients with[8 CTCs/7.5 ml blood had a significantly shorter survival time than the remaining patients with\8 CTCs after therapy [3]. Hou et al. drew the same conclusion using 50 CTCs/7.5 ml blood as the cut-off value, citing an overall survival of 5.4 versus 11.5 months for the high CTC cohort [4]. More studies using standardized methods are needed to validitate a correlation between CTC count and prognosis/disease response. In the latter study, CTCs were present in 85 % of patients and were abundant (range 0–44,896, mean ± SD: 1589 ± 5565), although a significant subpopulation of the CTCs was apoptotic [4]. Molecular characterization of CTCs may provide novel insights into progression of SCLC but investigations are hampered by insufficient tissue for research, because surgical resection and/or serial biopsies are rarely performed. In order to identify the functional properties of different CTC subsets, expansion of CTCs in cell culture systems is required. First ex vivo culture of CTCs has been described in breast cancer by Zhang et al. and Yu et al. [5]. However, these cultures required very high CTC concentrations in suitable blood samples ([1000 cells/7.5 ml in breast cancer), which were so far only found in a few patients. In colon cancer, CTCs isolated from the blood of one patient with a rapidly progressing tumor (out of 71 patients) displaying a CTC count[2300 cells/7.5 ml blood gave rise to a stable colon CTC line [6]. Another approach to expand the number of CTCs is xenotransplantation of patientderived CTCs into immunodeficient mice. Only blood & Gerhard Hamilton gerhard.hamilton@toc.lbg.ac.at

AIMS Nuclear imaging may have an increasing role in the diagnosis of hepatocellular carcinoma (HCC). The aim of this study was to compare prospectively the Gallium-67 citrate ((67)Ga) scintigraphy results with those obtained by positron emission tomography (PET) using (11)C-acetate in patients with HCC. METHODS We prospectively analysed 21 patients (mean age, 64+/-11 years) with histopathologically verified HCC undergoing (11)C-acetate PET and (67)Ga scintigraphy. (67)Ga scans were not performed in three of these 21 patients due to the exacerbation of the disease. Whole-body (11)C-acetate PET were performed following intravenous injection of 850 MBq of (11)C-acetate. For (67)Ga scintigraphy, whole-body, planar and single photon emission computed tomography imaging acquisitions were performed after intravenous application of a mean dose of 189 MBq (67)Ga. RESULTS (67)Ga scintigraphy found abnormalities only in 10 of 18 patients (56%) and detected 22 of 46 clinically involved sites (48%); it was false-positive in two patients. (11)C-acetate PET found abnormalities in 14 of 18 patients (78%) and detected 36 of 46 clinical lesions (78%); it was false-positive in one patients. In one patient with left supraclavicular lymph node metastases, neither the (67)Ga scintigraphy nor the conventional computed tomography have shown the lesions, which were clearly demonstrated by the (11)C-acetate PET. CONCLUSION Our results indicate significantly higher sensitivity and specificity of (11)C-acetate PET than (67)Ga scan in detection of HCC lesions. This study suggests that imaging with (11)C-acetate PET might play a potential role in the diagnostic workup of patients with HCC.

Small cell lung cancer (SCLC) is distinguished by excessive numbers of circulating tumor cells (CTCs) in extended and recurring disease. This malignancy has a poor prognosis due to rapid emergence of chemoradioresistant relapses after first-line chemotherapy. In vitro expansion of several CTC lines allowed for a detailed study of the contribution of these cells to metastasis. Generation of CTCs involves the establishment of co-cultures and recruitment of macrophages and specific cytokines. All cell lines show E-cadherin-positive epithelial-like cells and spontaneous assembling into very large tumorospheres. Such multicellular aggregates seem to be responsible for the observed broad resistance due to limited access to drugs, quiescent cell layers, and hypoxic cores.

Primary pulmonary hypertension is a fatal disease causing progressive right heart failure within 3 years after diagnosis. We describe a new concept for treatment of the disease using vasoactive intestinal peptide, a neuropeptide primarily functioning as a neurotransmitter that acts as a potent systemic and pulmonary vasodilator. Our rationale is based on the finding of a deficiency of the peptide in serum and lung tissue of patients with primary pulmonary hypertension, as evidenced by radioimmunoassay and immunohistochemistry. The relevance of this finding is underlined by an upregulation of corresponding receptor sites as shown by Northern blot analysis, Western blot analysis, and immunological techniques. Consequently, the substitution with the hormone results in substantial improvement of hemodynamic and prognostic parameters of the disease without side effects. It decreased the mean pulmonary artery pressure in our eight study patients, increased cardiac output, and mixed venous oxygen saturation. Our data provide enough proof for further investigation of vasoactive intestinal peptide and its role in primary pulmonary hypertension.

Lymphoma of the mucosa-associated lymphoid tissue (MALT) type usually arises in MALT acquired through chronic antigenic stimulation triggered by persistent infection and/or autoimmune processes. Due to specific ligand–receptor interactions between lymphoid cells and high-endothelial venules of MALT, both normal and neoplastic lymphoid cells display a pronounced homing tendency to MALT throughout the body. In the case of neoplastic disease these homing properties may be responsible for lymphoma dissemination among various MALT-sites. According to this concept, we have standardized staging procedures in all patients diagnosed with MALT-type lymphoma. All patients with MALT-type lymphoma underwent standardized staging procedures before treatment. Staging included ophthalmologic examination, otolaryngologic investigation, gastroscopy with multiple biopsies, endosonography of the upper gastrointestinal tract, enteroclysis, colonoscopy, computed tomography of thorax and abdomen and bone marrow biopsy. Biopsy was performed in all lesions suggestive for lymphomatous involvement, and evaluation of all biopsy specimens was performed by a reference pathologist. 35 consecutive patients with histologically verified MALT-type lymphoma were admitted to our department. Twenty-four patients (68%) had primary involvement of the stomach, five (15%) had lymphoma of the ocular adnexa, three (8.5%) had lymphoma of the parotid, and three (8,5%) of the lung. Lymph-node involvement corresponding to stage EII disease was found in 13 patients (37%), only one patient with primary gastric lymphoma had local and supradiaphragmatic lymph-node involvement (stage EIII). Bone marrow biopsies were negative in all patients. Overall, eight of 35 patients (23%) had simultaneous biopsy-proven involvement of two MALT-sites: one patient each had lymphoma of parotid and lacrimal gland, conjunctiva and hypopharynx, conjunctiva and skin, lacrimal gland and lung, stomach and colon, and stomach and lung. The remaining two patients had bilateral parotideal lymphoma. Staging work-up was negative for lymph-node involvement in all of these eight patients. The importance of extensive staging in MALT-type lymphoma is emphasized by the demonstration of multiorgan involvement in almost a quarter of patients. In addition, our data suggest that extra-gastrointestinal MALT-type lymphoma more frequently occurs simultaneously at different anatomic sites than MALT-type lymphoma involving the GI-tract. © 2000 Cancer Research Campaign

BACKGROUND In an intermittent androgen suppression therapy (IAS) trial we observed regular cycles of tumor suppression and regrowth in the majority of patients (17 +/- 2.6 month), with a subpopulation of patients (14 of 72) exhibiting a prolonged response of 28 +/- 7 month (range 18-64+ month) to the first eight-month androgen suppression cycle. PURPOSE To compare clinical data and laboratory tests (testosterone, prostate-specific antigen, and tissue polypeptide-specific antigen) of matched IAS patients showing either regular treatment cycles (n = 16) or prolonged response (n = 14). RESULTS Periods of androgen suppression resulted in reversible reduction of serum testosterone (< 1 nmol/l), PSA (< 1 ng/ml) indicating partial growth arrest and apoptotic regression of the tumors. The long-term response subgroup showed significantly lower mean values of tumor gradings, of pretreatment PSA values (11.36 +/- 4.54 vs. 47.5 +/- 12.4 ng/ml PSA), of PSA nadirs during androgen suppression (0.5 vs. 1 ng/ml PSA), and of overall testosterone values (3.9 +/- 1.14 vs. 6.6 +/- 1.18 mmol/l pretreatment) associated with low TPS values. CONCLUSIONS Patients exhibiting prolonged response to a single cycle of androgen suppression during IAS are characterized by a lower tumor burden, slow growing and less aggressive tumors, lower testosterone serum concentrations and lower absolute PSA nadirs during androgen suppression compared to a matched control group of short-term responders.

Long acting somatostatin-14 (SST) analogs are used clinically to inhibit tumor growth and proliferation of various tumor types via binding to specific receptors (R). We have developed a 111In-/90Y-labeled SST analog, DOTA-(d)βNal1-lanreotide (DOTALAN), for tumor diagnosis and therapy. 111In-/90Y-DOTALAN bound with high affinity (dissociation constant, Kd, 1–12 nm) to a number of primary human tumors (n = 31) such as intestinal adenocarcinoma (n = 17; 150-4000 fmol/mg protein) or breast cancer (n = 4; 250-9000 fmol/mg protein). 111In-/90Y-DOTALAN exhibited a similar high binding affinity (Kd, 1–15 nm) for the human breast cancer cell lines T47D and ZR75–1, the prostate cancer cell lines PC3 and DU145, the colonic adenocarcinoma cell line HT29, the pancreatic adenocarcinoma cell line PANC1, and the melanoma cell line 518A2. When expressed in COS7 cells, 111In-DOTALAN bound with high affinity to hsst2 (Kd, 4.3 nm), hsst3 (Kd, 5.1 nm), hsst4 (Kd, 3.8 nm), and hsst5 (Kd, 10 nm) and with lower affinity to hsst1...

BACKGROUND The present study evaluated monthly measurements of free and total prostate-specific antigen (PSA), and the tumor proliferation markers tissue polypeptide-specific antigen (TPS) and cytokeratin fragment 21-1 (CYFRA 21-1) in patients with advanced prostate cancer receiving intermittent androgen suppression therapy (IAS). METHODS Thirty-four men received alternating cycles of 8 month androgen suppression and treatment cessation (mean duration, 10.3 months) until PSA increased to >20 μg/l. Measurements of testosterone, percentage of free PSA, TPS, and CYFRA 21-1 were performed using ELISA and RIA assays. RESULTS Periods of androgen suppression resulted in reversible reductions of testosterone (from 6 ± 0.8 to

The excessive uncontrolled activation of inflammatory cells and mediators after trauma or major surgery plays a key role in the development of adult respiratory distress syndrome and multiple organ system failure (MOSF). In the past elevated cytokine levels were shown to influence the outcome of these patients adversely. There are diverging results regarding the removal of circulating cytokines by various methods of hemopurification for clinical improvement of MOSF. Seven patients after trauma or major surgery underwent continuous venovenous hemofiltration (CVVH) for the treatment of severe organ failure of the heart and lungs (Murray score 2.74) but not for renal or liver failure. The cytokine levels were measured at the beginning and 15, 60, 120, and 240 minutes after initiation of CVVH (measure points MP1–5). Clinical improvement during the treatment was monitored, and correlation with cytokine levels was evaluated. Arterially measured tumor necrosis factor α rose from 11.14 ng/ml to 17.86 ng/ml (p < 0.05). Arterial interleukin-6 (IL-6) levels significantly decreased during CVVH from 1284.7 ng/ml to 557.9 ng/ml; IL-8 levels simultaneously decreased from an initial peak of up to 154.4 ng/ml at MP3 to 97.3 ng/ml at MP5. The drop in serum IL-6 and IL-8 levels closely correlated with clinical improvement. After 2 hours of CVVH the hemodynamic situation improved significantly, as revealed by a decrease in catecholamine expenditure, an increase in arterial pressure, and a decrease in pulmonary artery pressure. Moreover, 2 hours after the initiation of CVVH the oxygenation index rose significantly and correlated well with the drop in shunt fraction. The Murray score significantly fell to 1.86. The removal of IL-6 and IL-8 by CVVH after initial stimulation correlates with clinical improvement, which was demonstrated by significantly improved oxygenation and hemodynamics from 2 hours after the initiation of CVVH onward. The elimination of cytokines and several mediators by CVVH may contribute to the cardiopulmonary improvement of critically ill patients. In comparison with the clinical control group ( n = 7), which was comparable in terms of MOSF, no intervention led to a similar improvement in cardiorespiratory failure, and overall two of these patients died. Moreover, patients of the control group experienced a significant longer stay at in the intensive care unit.

Grundlagen: Die schnelle Restitution ermoglicht den raschen Schlus von Epitheldefekten durch Enterozytenmigration. Wir untersuchten den Einflus extrazellularer Matrixproteine auf die Restitution der sauregeschadigten Mukosa des Kaninchenduodenums in vitro. Methodik: Schleimhautpraparationen des Kaninchenduodenums wurden in Ussing-Kammern eingebracht und nach einer 30minutigen Aquilibrierungsphase durch 10 min luminale Exposition mit 10 mM HCl geschadigt. Danach wurden die Gewebe 3 Stunden mit Puffer (Kontrollen) oder Puffer, der auf der luminalen Seite die Testsubstanzen enthielt, inkubiert (n=6 pro Gruppe). Potentialdifferenz und Kurzschlusstrom wurden gemessen, der Widerstand (R) nach dem Ohmschen Gesetz berechnet. Nach Versuchsende wurden HE-gefarbte histologische Schnitte hergestellt, das Ausmas der Schadigung mit einer speziellen Morphometrieanlage gemessen. Ergebnisse: Nach Schadigungsende waren 58±4%, nach 1 und 3 h 40±4% bzw. 36±2% geschadigt (Kontrollen, n=6). Nach HCl-Exposition losten sich die geschadigten Enterozyten der oberen 2 Zottendrittel ab, so das luminal denudierte Stummelzotten resultierten, welche 3 h nach Schadigung teilweise von flachen Enterozyten bedeckt waren. Luminales Rinderserumalbumin (38±5%), Antikorper gegen Kollagen IV (35±7%) und Fibronektin (38±4%) sowie jeweils 25, 50 und 100 µg/ml Kollagen IV (34±3, 35±2 bzw. 33±4%) und Fibronektin (39±2, 36±4 bzw. 36±2%) hatten keinen Einflus auf die Restitution (n=6). Antilaminin fuhrte zu einer signifikanten Zunahme der Schadigung auf 64±5% (p

Grundlagen: Clostridium difficile ist der Erreger der Antibiotika-assoziierten Enterokolitis. In Tierstudien schadigt Clostridium difficile Toxin A, aber nicht Toxin B die Schleimhaut des Gastrointestinaltraktes. Die Wirkung der Toxine auf die humane Kolonschleimhaut ist unbekannt. Deshalb untersuchten wir in dieser Studie die Wirkung von Clostridium difficile Toxin A (TxA) und Toxin B (TxB) auf die humane Kolonschleimhaut in vitro.

Growth factors are mainly involved in the regulation of intestinal epithelial barrier function. This study investigated the effect of epidermal growth factor (EGF) and insulin-like growth factor 1 (IGF-1) on epithelial restitution of rabbit duodenum in vitro.Rabbit duodenal mucosal strips mounted in an Ussing chamber were luminally exposed to 10 mmol/L HCl for 10 minutes and then incubated with buffer alone or luminal buffer containing various concentrations of EGF and IGF-1 for 3 hours. Resistance was calculated from potential difference and short-circuit current. Damage was assessed by morphometry on HE-stained sections.HCl caused resistance to decrease from 112 +/- 2 to 51 +/- 4 ohms x cm2 10 minutes after injury (n = 6; P0.05). Postinjury treatment with 25 or 50 ng/mL luminal EGF for 3 hours stimulated resistance to recover to 94 +/- 3 and 104 +/- 3 ohms x cm2, respectively, vs. 81 +/- 3 omega x cm2 in controls (P0.05). Ten minutes after injury, 62% of the mucosa was damaged; 3 hours after injury, damage was reduced to 24% +/- 1.09% and 10% +/- 1.42% in the 25 and 50 ng/mL EGF group, respectively, vs. 38% +/- 0.93% in controls (n = 6 per group). EGF stimulated enterocyte migration. IGF-1 did not impair epithelial restitution.EGF, but not IGF-1, promoted epithelial restitution of rabbit duodenum in vitro.

Grundlagen Die Multidrug-Resistenz (MDR) gilt als wichtiger Chemoresistenzmechanismus besonders bei hamatologischen, aber auch bei soliden Malignomen. Inhibitoren des MDR-Effektors P-Glycoprotein (P-GP), Chemomodulatoren genannt, zeigten fur den Fall solider Tumoren aufgrund nebenwirkungsbedingter Einschrankungen der erreichbaren Konzentrationen bisher noch keine klinischen Erfolge. Mit der Fragestellung, ob und wie weit Chemomodulatoren den P-Gpmediierten Zytostatikatransport in intestinalen Karzinomzellen hemmen, wurden neben dem Antiostrogen Tamoxifen (TMX) die speziell fur die MDR-Reversion entwickelten Chemomodulatoren, namlich das weniger kardiotoxische R-Enantiomer von Verapamil (R-VPL) und das nichtimmunsuppressive Cyclosporin SDZ PSC-833 (PSC), in vitro getestet. Methodik Unter Verwendung von HCT-8 intestinalen Adenokarzinomzell-Monolayem untersuchten wir den P-Gp-mediierten, transmembranen, sekretorischen Transport von Vinblastin, einem P-Gp-Substrat, sowie den zellassoziierten Vinblastingehalt in An- oder Abwesenheit obengenannter Chemomodulatoren unter verschiedenen extrazellularen pH-(pHo-)Bedingungen, wie sie in soliden Tumoren vorkommen. Zusatzlich fuhrten wir durchfluszytometrische Effluxmessungen mit dem P-Gp-Substrat Rhodamin 123 unter denselben extrazellularen Bedingungen durch. Das Ausmas der durch die Chemomodulatoren bedingten Resistenzreversion wurde anhand von Chemosensitivitatsassays (Inkorporation von3H-Thymidin) mit HCT-8 Einzelzellsuspensionen gezeigt. Ergebnisse Die Modulatoren R-VPL, PSC und TMX inhibierten den transepithelialen VIN-Transport in einem Ausmas von bis zu 50,55 bzw. 30%, der zellassoziierte Vinblastingehalt war nach 5h im Vergleich zur Kontrolle 3,7-, 2,3- bzw. 1.7mal groser. Im Gegensatz zu PSC und TMX stellten wir bei Verwendung von R-VPL einen 32-bzw. 47%igen Verlust der modulierenden Wirkung bei extrazellularer Ansauerung (pHo 7,0 bzw. 6,8) fest. Damit ubereinstimmende Ergebnisse erzielten wir auch bei den durchfluszytometrischen Studien unter gleichen extrazellularen Bedingungen. Die Chemosensitivitatsassays ergaben einen 8-, 5.3- und 10,7fachen Anstieg der VIN-Toxizitat durch R-VPL (10 μM), PSC (100 ng/ml) bzw. TMX (10 μM). Schlusfolgerungen Mit TMX konnte trotz schwacher P-Gp-Inhibition eine deutliche Resistenzreversion bewirkt werden, was darauf hindeutet, das zusatzliche Mechanismen an der Resistenz der Zellinie beteiligt sind, die durch TMX, unabhangig von P-Gp-Interaktionen, beeinflusbar sind. Der festgestellte Wirkungsverlust von R-VPL unter sauren Bedingungen konnte als Erklarung der vorherrschend negativen Ergebnisse klinischer Studien mit Verapamil bei Tragern solider Tumoren dienen. Sofern MDR wesentlich zum fehlenden Ansprechen solider Tumoren auf Chemotherapie beitragt, wurden wir klinische Erfolge mit PSC erwarten, das sich bei geringer klinischer Toxizitat durch ein hohes vom pHo weitgehend unabhangiges P-Gp-Inhibitions-potential auszeichnet.

Grundlagen: Resistenzmechanismen auf Tumorebene und zellularer Art sind fur die niedrigen Ansprechraten und Uberlebenszeiten nach Chemotherapie, insbesondere bei den meisten soliden Tumoren, hauptverantwortlich. Die sogenannte Multidrug-Resistenz (MDR) wird durch ein Transmembranprotein (P-Gp) bewirkt, das klinisch wichtige Zytostatika aus dem Zellinneren transportiert und in hamatologischen und soliden Tumoren nachgewiesen wurde. P-Gp wird im Normalgewebe in Epithelien der Nierentubuli, der Leber, der Nebenniere, der Blut-Hirnschranke und andernorts exprimiert. Es war das Ziel der vorliegenden Arbeit, die Funktion des P-Gp und seine Inhibierbarkeit durch Chemomodulatoren, im Gegensatz zu den normalerweise untersuchten Suspensionszellen, in polarisierter, apikaler Expression eines Epithelmodells zu erfassen. Als Zellsystem wurden Monolayerkulturen der humanen HCT-8-Adenokarzinomlinie auf porosen Gewebekulturinserts etabliert.

Hintergrund: Solide Tumoren weisen azidotische Regionen mit durchschnittlichen extrazellularen pH Werten (pHo) von 6,9 bis 7,0 (Normalgewebe: 7,4 bis 7,5) auf. In humanen SW620-Kolonkarzinomzellen konnten in einer fruheren Arbeit ein Na/H-Austauscher sowie ein Na/HCO3-Kotransporter als Saureextrusoren nachgewiesen werden, ein Cl/HCO3-Austauscher wurde im Gegensatz zu anderen Tumorlinien nicht gefunden. Diese Studie untersucht die Auswirkungen unterschiedlicher extrazellularer pH-Bedingungen auf die intrazellulare pH-Regulation am Modell der SW620-Kolonkarzinomzellen, durch Variation des CO2-Partialdrucks und der HCO3-Konzentration der Inkubationslosungen.

Prostate cancer is among the most common types of malignancies and causes of cancerrelated deaths in men worldwide (Fitzpatrick, et al., 2009). Primary tumor involvement outside the prostatic capsule or relapse following radical prostatectomy results generally in incurability (Lassi & Dawson, 2009). Androgen deprivation therapy has progressed since attempts in 1941, when surgical castration had been shown to improve outcomes for the first time. Palliative treatment consists of hormonal manipulation to deprive the cancer cells of androgenic stimulation by orchidectomy or use of LHRH analogs and steroidal or nonsteroidal antiandrogens (Kollmeier & Zelefsky, 2008). Although continuous androgen suppression therapy (CAS) has been a cornerstone of the management of prostate cancer for more than 50 years, controversy remains regarding its optimum application. Generally, androgen suppression (AS) is performed as continuous treatment, resulting in apoptotic regression of the tumor cells in a high percentage of cases. However, surgical or medical castration results in median progression-free survival of only 2–3 years, with no other effective treatment left (Mellado, et al., 2009). Responses to cytotoxic therapy are low and only recently several studies revealed a possible benefit of incorporating chemotherapeutic agents in treatment regimen for prostate cancer (Chang & Kibel, 2009). Taxanes like docetaxel and cabazitaxel, therapeutic cancer vaccines and newly developed agents targeting androgen receptor signaling are expected to improve therapy (Madan et al., 2011). Following experimental research using animal models, intermittent androgen suppression (IAS) was introduced as new clinical concept, assuming that during limited regrowth in the treatment cessation periods tumorigenic cells are residing in an androgen-responsive state (Goldenberg, et al., 1995). Since induction of androgen independence may occur early after treatment initiation, cessation of antiandrogen therapy prior to this switch is expected to maintain the apoptotic potential of the tumor cells and keep them sensitive to retreatment. Providing an easy method for selection of the type of treatment and early assessment of tumor growth during the off-periods serial serum PSA determinations made IAS feasible. In detail, IAS consists of an initial androgen suppression period of up to nine months combining LHRH antagonists and antiandrogens, which is followed by treatment cessation

Pancreatic adenocarcinoma is a devastating disease characterized by early dissemination and poor prognosis. These solid tumors express receptors for neuropeptides like neurotensin (NT) or epidermal growth factor (EGF) and exhibit acidic regions when grown beyond a certain size. We previously demonstrated increases in intracellular Ca2+ levels, intracellular pH and interleukin-8 (IL-8) secretion in BxPC-3 and PANC-1 pancreatic cancer cells in response to a stable NT analog. The present study aimed at investigation of the dependence of the relative expression of NT receptor 1 (NTR1) and EGFR in BxPC-3 and MIA PaCa-2 cells on cell density and extracellular pH (pHe). MTT assays revealed the NTR1 inhibitor SR 142948-sensitive Lys8-ψ-Lys9NT (8–13)-induced proliferation in BxPC-3 and PANC-1 cells. Confluent cultures of BxPC3 and HT-29 lines exhibited highest expression of NTR1 and lowest of EGFR and expression of NTR1 was maximal at slightly acidic pHe. IL-8 production was stimulated by Lys8-ψ-Lys9NT (8–13) and even enhanced at both acidic and alkaline pHe in BxPC-3 and PANC-1 cells. In conclusion, our in vitro study suggests that one contributing factor to the minor responses obtained with EGFR-directed therapy may be downregulation of this receptor in tumor cell aggregates, possibly resulting in acquisition of a more aggressive phenotype via other growth factor receptors like NTR1.

The treatment of advanced metastatic prostate cancer by hormone manipulation or orchiectomy is frequently followed by the appearance of hormone-insensitive and highly chemoresistant tumor cells. In this study we have investigated the contribution of the P-glycoprotein-mediated drug efflux (multidrug-resistance; MDR) to the cellular resistance of prostate carcinoma-derived cell lines to diverse cytotoxic drugs by detection of P-glycoprotein (P-gp) measurement of P-gp-mediated drug transport and reversal of MDR by chemosensitizers. The in vitro chemosensitivity of three prostate cancer cell lines (PC-3, DU-145 and LNCaP) to doxorubicin was measured in a thymidine incorporation proliferation assay. Growth of the partially hormone-sensitive cell line LNCaP is inhibited by low doses of doxorubicin (IC50:27 ng./ml.), but PC-3 and DU-145 are highly resistant to the drug, with IC50 values of 10 micrograms./ml. and 7.5 micrograms./ml., respectively. The chemosensitivity of the PC-3 and DU-145 cells is increased in response to 1 microM. verapamil, 1 micrograms./ml. cyclosporine A and 2 microM. tamoxifen, which are known to partially reverse the MDR phenotype in other resistant tumors. A verapamil-sensitive drug efflux has been demonstrated for the PC-3 and Du-145, but not for the LNCaP, cell lines, using flow cytometric measurements of the P-gp substrate rhodamine 123 efflux from preloaded cells. In agreement with the functional measurements, the expression of the P-glycoprotein was detected in the PC-3 and Du-145 cell lines in Western blots using the monoclonal C 219 antibody. In conclusion, the chemoresistant and hormone-insensitive PC-3 and Du-145 cell lines express P-gp and exhibit verapamil-sensitive drug efflux, indicative of MDR. However, the low MDR-reversal rates observed in these cell lines in response to chemosensitizers in clinically achievable concentrations (approximately 2- to 3-fold reversal), point to non-MDR-associated cellular mechanisms as dominant factors of chemoresistance in prostate cancer.

In human orthotopic liver transplantation (LTX) intraoperative elevations of TNF-α (> 100 pg/ml) and IL-6 (>800 pg/ml) have been found to correlate with early post-operative rejections and infections respectively. In this study the possible mechanism responsible for the induction of these cytokines has been investigated during liver allografting in 38 recipients. Intraoperative elevations of TNF-α (> 100 pg/ml) were detected in the majority of pre-transplant endotoxin positive recipients (8/12, > 10 endotoxin units/ml), the patients turning endotoxin positive until the end of grafting (3/5), and in a subgroup (6/21 patients), apparently endotoxin negative for the whole operation. Therefore endotoxin (ET) seems to stimulate release of TNF-α in approximately 50% of the patients, whereas sensitized Kupffer graft cells or immediate allograft reactivity of the host are likely to account for the remaining TNF-α positive cases. Elevations of IL-6 > 800 pg/ml) were found in approximately 50% of the TNF-α positive cases, indicating partially independent regulatory pathways for IL-6 induction in the TNF-α negative patients. In agreement with a previous study, 11/13 (85%) of the intraoperative TNF-α positive recipients rejected their grafts within the first 10 days post-operatively. These data demonstrate that ET/infection associated as well as ET independent/reperfusion associated intraoperative TNF-α elevations, promote the initiation of allograft rejection in human liver transplantation. The transient and low endotoxaemia caused by the liver grafting procedure performed without veno-venous bypass seems to be of minor importance in the intraoperative induction of TNF-α.

Acidosis commonly observed in solid tumors like pancreatic cancer promotes genetic instability and selection of a more malignant phenotype of cancer cells. Overexpression or activation of integral membrane proteins mediating H+ efflux may contribute to extracellular acidification. Neurotensin (NT) induces intracellular alkalinization and stimulates interleukin-8 production in pancreatic cancer cells and, as demonstrated here, the stable NT analog Lys(8)-psi-Lys(9)NT(8-13) enhances the amiloride-sensitive, Na+-dependent transmembrane H+ flux by a factor of 2.05+/-0.28 and 2.69+/-0.07 in BxPC-3 and PANC-1 pancreatic cancer cells, respectively, by phosphorylation of the Na+/H+ exchanger 1 (NHE1). Human genome-wide gene expression analysis was performed to detect effects of Lys(8)-psi-Lys(9)NT(8-13) on BxPC-3 cells. Results indicated upregulation of genes involved in regulation of NHE1, hypoxic response and glycolysis in response to Lys(8)-psi-Lys(9)NT(8-13) even under normoxic conditions. Therefore, our findings suggest that growth factors like NT may be implicated in the early progression of pancreatic cancer by localized acidification and induction of an aerobic glycolytic phenotype with higher metastatic potential in small cell aggregates.

IMPORTANCE IN THE FIELD: Primary bone tumors are considered as (sex steroid) hormone-dependent tumors. Osteosarcoma, osteoblastoma and bone cysts are preferentially found in males, while giant cell tumors are more common in females. Indeed, bone tumor development and progression are influenced by sex steroid hormones derived from in situ synthesis in bone cells.This review describes intracrine mechanisms for local formation of the biologically most active estrogen, 17beta-estradiol (E2), from circulating steroid precursors through the 'aromatase' (aromatization of androgens) and the 'sulfatase' (conversion of inactive estrone-sulfate) pathway.The reader gains knowledge on both pathways and the enzymes, which contribute to the in situ availability of active hormones, namely 3beta-hydroxysteroid dehydrogenases, 17beta-hydroxysteroid dehydrogenases, aromatase, steroid sulfatases and sulfotransferases. An overview is given and the expression and function of these enzymes in bone tumors are discussed.Knowledge on pathways for the in situ formation of E2 in bone cells may allow the identification of potential targets for i) novel endocrine therapeutic options in primary bone tumors and ii) future preventive interventions.

Im Verlauf einer gramnegativen abdominellen Sepsis wurden prospektiv bei 18 Patienten Endotoxin, TNFa und IL-6 im Plasma gemessen und mit dem Uberleben sowie der Lungen- und Leberfunktion korreliert. Wahrend Endotoxin (median 12 vs. 9 EU/ml) und TNFa (12 vs. 21 pg/ml) bei Uberlebenden und Toten nicht unterschiedlich waren, fand sich eine signifikante Erhohung von IL-6 bei letalem Verlauf (82 vs. 171 pg/ml, p=0,02). An Tagen mit Lungenversagen war IL-6 signifikant erhoht (70 vs. 159 pg/ml, p=0,009). Endotoxin (11 vs. 13 EU/ml) und TNFa (12 vs. 15 pg/ml) waren bei Patienten mit und ohne respiratorischer Insuffizienz nicht different. Ebenso war IL-6 bei Leberversagen signifikant hoher (349 vs. 69 pg/ml, p=0,0000), wahrend fur Endotoxin (11 vs. 13 EU/ml) und TNFa (33 vs. 17 pg/ml) kein Unterschied nachgewiesen werden konnte. Bei gramnegativer abdomineller Sepsis fand sich eine Korrelation von IL-6 mit Uberleben, Lungen- und Leberfunktion, wahrend Endotoxin und TNFa in diesen Parametern nicht signifikant waren.

The correlation of endotoxin (ET), tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and cellular immune parameters with multiple organ failure and lethal outcome in intraabdominal infections was studied in a group of 18 patients with peritonitis, abscess or pancreatitis. Of these patients, 7 developed respiratory failure and 5 died due to multiple septic organ failure. The peak levels of ET (2.7 +/- 1.3 ng/ml) in the course of the disease were followed by moderate increases of TNF-alpha (mean 147 +/- 41 pg/ml) and IL-6 (170 +/- 61 pg/ml) within 2 days. Analysis of the parameters for the last 12 days prior to death or discharge showed, that the patient group with lethal outcome was characterized by significant lower mean plasma levels of TNF-alpha (less than 75 pg/ml versus greater than 160 pg/ml) and IL-6 (less than 130 pg/ml versus greater than 270 pg/ml), as well as high rates of unstimulated thymidine uptake into peripheral mononuclear blood cells (greater than 44000 cpm/8 x 10(6) PMBC/18 h versus less than 24000 cmp), T-lymphocyte depression (CD3; approximately greater than 40% reduction) with lower T-helper/inducer subset cell numbers (mean CD:CD8 ratio 1.0 +/- 0.55 versus 1.8 +/- 0.2) and lower lectin (PHA) stimulation values (1.9 +/- 1.4 versus 4.1 +/- 1.0). These data demonstrate an anergic immune status with low mediator levels and depressed T-lymphocyte function in patients with poor prognosis.(ABSTRACT TRUNCATED AT 250 WORDS)

In cancer patients detection of systemic disease is of great importance to obtain prognostic information and to guide therapy. Bone marrow (BM) seems to be a common homing tissue for the early spread of tumor cells from various epithelial tumors; however, verification of the prognostic significance of BM-disseminated tumor cells (BM-DTCs), is restricted to breast cancer so far. These cells may be dormant for a long time, and signals triggering their activation leading to recurrence remain to be characterized. A recent study involving metastatic breast cancer patients reported that the shortest disease-free survival is correlated with cytokeratin (CK)-negative BM aspirates and that CK-positive BM-DTCs correspond to dormant tumor cells. Soluble CK fragments in serum including CK18 and 19 (measured as TPS and CYFRA 21-1, respectively) and caspase-cleaved CK18 are widely used to monitor tumor progression and response to therapy, actually indicating proliferation and/or necrotic/apoptotic cell death. In order to assess the source of the CK fragments, we used determinations of CK18 and caspase-cleaved CK18 fragments in serum samples before and after radical tumor surgery in colon cancer patients. Elevated serum concentrations of CK18 were found to persist in patients with a high incidence of BM-DTCs, and high perioperative levels of caspase-cleaved CK18 fragments were detected in patients with early relapses, respectively. These results indicate that in some patients at increased risk of recurrence disseminated cell populations exist that are responsible for the release of the bulk of CK fragments after removal of the apparently nonmetastatic tumor. In good agreement with the results in metastatic breast cancer patients, release of CK18 or 19 fragments by BM-DTCs seem to indicate disseminated tumor cells mainly in a dormant state, whereas caspase-cleaved CK18 may indicate skipping of this latent phase and early progression. Therefore, caspase cleavage of CKs in intact tumor cells seems to accompany or is involved in the differentiation leading from dormant to progressively active disseminated tumor cells. Release of respective CK fragments would result in an apparent clearing of CK-positive cells in BM, leaving malignant cells that have possibly undergone an epithelial-mesenchymal transition. Micrometastatic cancer cell lines derived from breast cancer patients were found to display loss of epithelial CK8, 18 and 19 as well as ectopic expression of vimentin as in mesenchymal cells. In conclusion, degradation of CKs may represent a marker indicating reactivation of dormant tumor cells in BM.

Pancreatic adenocarcinomas express neurotensin receptors in up to 90% of cases, however, their role in tumor biology and as a drug target is not clear. In the present study, a stable neurotensin (NT) analog induced intracellular calcium release and intracellular alkalinization in BxPC-3 and PANC-1 pancreatic cancer cells that was abolished by inhibitors of NT receptor (NTR) and sodium-proton exchanger 1 (NHE1), amiloride and SR 142948, respectively. Activation of NHE1 involved increased phosphorylation of dimethylfumarate-sensitive mitogen- and stress-activated kinase 1/2 (MSK1/2). NTR signaling appears to promote a metastatic phenotype in pancreatic cancer cells by induction of localized extracellular acidification in normoxic cells, preceeding acidosis induced by hypoxia and switch to glycolysis in addition to increased expression of interleukin-8 (IL-8).

Background Soluble cytokeratin 18 (CK18; M65) and a caspase-cleaved fragment of CK18 (M30) have been used as biomarkers, corresponding to tumor cell death and apoptosis, respectively. Methods In the present study, M30 was quantified for the first time in serum samples of colon cancer patients pre- and postoperatively as well as during chemotherapy. Minimal residual disease (MRD) was assessed preoperatively by detection of pan-cytokeratin antibody A45-B/B3-positive cells in bone marrow aspirates. Results Out of 46 patients, those with colon tumors of stages I and IV had significantly elevated M30 serum concentrations compared to controls (n = 23). In 31 colon cancer patients, M30 determinations were performed prior to and seven days after tumor surgery. A group of 24 patients exhibited a significant decrease of M30 in response to tumor removal, in contrast to seven patients who revealed either persistent or higher M30 levels postoperatively. The frequency of MRD was not significantly different for patients with decreasing (4/24) and persisting (3/7) M30. However, M30 correlated significantly with the increased number of recurrences within 36 months in the group with persisting M30 (4/7 versus 2/24, p = 0.032; hazard ratio 8.3, p = 0.016). In a group of patients (n = 10) receiving capecitabine/oxaliplatin chemotherapy (CapOx), transient increases in M30 did not correlate with responses. Conclusion The data obtained within the present limited pilot study in colon cancer patients demonstrate that perioperative changes of M30 may indicate systemic residual tumor load and increased risk of recurrence warranting further evaluation of this marker of apoptosis in a larger prospective clinical trial.

Fourteen patients with metastatic pancreatic adenocarcinoma were treated with the long-acting somatostatin (SST) analogue lanreotide. No objective response was obtained, and the median survival was 4 months (range 1.8–7 months). Pancreatic cancer could not be visualized by means of SST-receptor (R) scintigraphy in our patients. In vitro data also demonstrated absence of SSTR2 expression, suggesting pancreatic cancer not to be a potential target for treatment with SST analogues. © 1999 Cancer Research Campaign

Prostate carcinoma-derived factors induce a proliferative response in osteoblasts. The present study investigated the involvement of MAP kinase in the osteoblastic reaction of osteocytes and the response of 1alpha,25-hydroxy-vitamin D3 (1,25-vitD3)-pretreated osteoblasts. Conditioned media (CM) from prostate, colon, pancreatic, renal cell and breast cancer cell lines were tested on their proliferative activity using murine osteoblast-like MC3T3-E1 cells, MG63 human osteosarcoma cells and immortalized human osteoblasts (AHTO-7). Changes in osteoblastic activities of the supernantants were measured in the presence of MAP kinase inhibitors and following 1,25-vitD3-induced differentiation of the target osteoblasts. Supernatants of prostate cancer cells stimulated proliferation of osteoblasts in all three indicator cell lines, with AHTO-7 exhibiting the most significant correlation to human primary osteoblast cultures. 1,25-vitD3 induced the differentiation marker alkaline phosphatase (ALP) in MC3T3-E1 and AHTO-7, but only to a minor degree in MG63 cells. 1,25-vitD3-induced differentiation reduced the proliferative response to CM from several cell lines in MC3T3-E1 and MG63 to a minor degree, whereas in AHTO-7 cells the osteoblastic reaction was reduced for 2/4 pancreatic, 3/3 colon and 1/1 renal cancer CMs, however not for 3/3 prostate cancer CMs. Stimulation of AHTO-7 cells by CM from prostate cancer lines is inhibited significantly by MEK1 kinase inhibitor PD 98059 in contrast to CMs derived from other carcinomas, except ACHN renal cancer cells. The findings in the present study demonstrate that human AHTO-7 cells seem to represent a valid human system to monitor osteoblastic activity, especially in respect to 1,25-vitD3-induced differentiation. Vitamin D3-induced differentiation has no direct effect on prostate cancer-derived osteoblastic activity in the same cell line in vitro, which however, could be reversed by disruption of the signal transduction at the MAP kinase level, revealing a new target for the inhibition of prostate cancer-associated bone formation.

BACKGROUND Recent studies have shown that vascular endothelial growth factor (VEGF) receptor is overexpressed in vascular endothelial cells of various human tumours as well as in human tumour cells. The aim of this study was to evaluate the usefulness of scanning with VEGF(165) labeled with (123)I for tumor localisation in patients with gastrointestinal tumours. PATIENTS AND METHODS Human recombinant VEGF(165) was radiolabelled with (123)I by electrophilic radioiodination using the chloramine T method. [(123)I]VEGF(165) was administered intravenously [mean dose 184 +/- 18 MBq (

10-hydroxycamptothecin (HCPT), a natural analog of the alkaloid camptothecin (CPT), is a promising anticancer agent currently undergoing preclinical trials. Though HCPT is less toxic and more active in various human cancer cell lines and in animal tumor models than the clinically approved CPT-analog topotecan, little is known about its biotransformation products and their route of elimination. To investigate the metabolism and biliary excretion, livers of male Wistar rats were perfused with HCPT (5 microM). Bile and perfusate samples were collected for 60 min and quantified by reversed-phase high-performance liquid chromatography (HPLC). Besides HCPT, three metabolites, namely HCPT glucuronide (M1), hydroxyHCPT glucuronide (M2), and hydroxyHCPT (M3) could be identified by enzymatic hydrolysis with beta-glucuronidase and mass spectroscopy. Biliary secretion of HCPT and M1-M3 reached a peak secretion of 1532+/-124, 75+/-16, 5.8+/-1.6 and 2.1+/-0.5 pmoles/g liver.min, respectively, after 25 min. The total amount of HCPT and M1-M3 excreted into bile during the time of perfusion (60 min) was low and represented a mean of 9.9+/-3.2%, 0.44+/-0.17%, 0.041+/-0.010%, and 0.022+/-0.004% of the initial HCPT dose, respectively. In the perfusate, besides HCPT M1 and M2 but not M3 could be detected (maximum concentrations after about 20 min: 3248+/-210, 16.8+/-2.8 and 1.0+/-0.4 pmoles/g liver.min, respectively). The cumulative efflux of HCPT and M1 and M2 into the perfusate was 21.1+/-3.9%, 0.145+/-0.036% and 0.018+/-0.004% of the initial dose, respectively, indicating a preferable non-biliary secretion for HCPT and a predominant biliary elimination for conjugated HCPT biotransformation products. In conclusion, HCPT is biotransformed in a rat liver model to three metabolites, mainly excreted into bile, which may be of clinical relevance during cancer therapy.

Based on the fact that somatostatin (SST) analogs have given promising results for treatment of hepatocellular cancer, we performed both in vitro and in vivo investigations to define the role of a depot formulation of the long acting SST-analog lanreotide (LAN). A decrease of cells in the S-phase as compared to controls (p