Suppressive effect of bevacizumab on peritoneal dissemination from gastric cancer in a peritoneal metastasis model

Purpose

Vascular endothelial growth factor (VEGF) has been reported to enhance vascular permeability and angiogenesis in the abdominal wall, thereby contributing to peritoneal dissemination with malignant ascites. We conducted this experimental study to find out if bevacizumab, a humanized monoclonal antibody against VEGF, had a suppressive effect on peritoneal dissemination from gastric cancer, in an experimental nude mouse model of peritoneal metastasis.

Methods

Each mouse was treated with a single intraperitoneal (i.p.) injection of bevacizumab. Five mice were killed, and we measured their body weight, the mean number of tumor nodules, and the volume of ascites. We also extracted retroperitoneal tissues for histological examination, to count the frequency of mitosis, and to calculate the mitotic index. Another five mice were monitored until death, and their mean survival duration was calculated.

Results

The volume of ascites and the mitotic index were significantly lower in the therapy group than in the nontherapy group (P = 0.042 and P < 0.01, respectively). The survival curve of the therapy group was significantly higher than that of the nontherapy group (P = 0.005).

Conclusion

Bevacizumab may suppress peritoneal dissemination from gastric cancer.     

Vascular Endothelial Growth Factor Receptor 2 Gene Polymorphisms as Predictors for Tumor Recurrence and Overall Survival in Non-Small-Cell Lung Cancer

Purpose

VEGFR-2 gene displays several functional germline polymorphisms with impact on VEGFR-2 mediated angiogenesis. Our purpose was to evaluate VEGFR-2 polymorphisms as prognostic markers for tumor recurrence and overall survival (OS) in non-small-cell lung cancer (NSCLC).

Methods

In total, 209 Caucasian patients who had been surgically treated for NSCLC between 1996 and 2010 were included in this study. Genotyping of peripheral blood cells was performed by TaqMan^? genotyping assays or polymerase chain reaction for five VEGFR-2 polymorphisms. Chi- square test, Kaplan?CMeier estimator, and Cox regression hazard model were used to assess the prognostic value of VEGFR-2 polymorphisms.

Results

VEGFR-2+4422 (AC)_10?C14 polymorphism was identified as a positive prognostic marker for time to metastasis (11/12 vs. 11/11 (AC) repeats: hazard ratio (HR), 0.28; 95% confidence interval (CI), 0.11?C0.75; p?=?0.012) and OS (11/12 vs. 11/11 (AC) repeats: HR, 0.41; 95% CI, 0.21?C0.82; p?=?0.012) in squamous cell carcinoma. For adenocarcinoma, VEGFR-2?906 C>T (C/T vs. CC: HR, 0.19; 95% CI, 0.43?C0.82; p?=?0.027) and VEGFR-2?271 G>A (G/A vs. G/G: HR, 0.25; 95% CI, 0.07?C0.86; p?=?0.027) predicted longer time to local recurrence and VEGFR-2?906 C>T was a predictor for better OS (T/T vs. C/C: HR, 0.28; 95% CI, 0.09?C0.84; p?=?0.024).

Conclusions

VEGFR2 germline polymorphisms predict tumor recurrence and OS in NSCLC.     

Lipocalin-2 is Associated With a Good Prognosis and Reversing Epithelial-to-Mesenchymal Transition in Pancreatic Cancer

Background

Lipocalin-2 is a multifaceted modulator in cancer progression. Its clinical significance is not clear in pancreatic cancer. The purpose of this study was to investigate whether lipocalin-2 is associated with good prognosis by reversing epithelial-to-mesenchymal transition (EMT) in pancreatic cancer.

Methods

Lipocalin-2, E-cadherin, or vimentin expression was detected in 60 pancreatic adenocarcinoma specimens. Correlations between lipocalin-2 expression and EMT, the clinicopathologic characteristics, and prognosis were investigated. Whether pancreatic cancer cells’ migration and invasion (some characteristics of EMT) were affected by lipocalin-2 was also explored.

Results

High lipocalin-2 expression was significantly associated with a good prognosis in pancreatic cancer (p < 0.05). Overexpression of lipocalin-2 correlated with a lower extent of EMT (p < 0.05), increased E-cadherin expression (p < 0.05), decreased vimentin expression (p < 0.05), and reduced cancer cell migration and invasion in pancreatic cancer.

Conclusions

Lipocalin-2 may be considered an epithelial inducer, which may reverse EMT and predict a good prognosis in pancreatic cancer.     

MicroRNA-10a is Overexpressed in Human Pancreatic Cancer and Involved in Its Invasiveness Partially via Suppression of the HOXA1 Gene

Background

There is increasing evidence that microRNAs are differentially expressed in many types of cancers. Despite progress in analyses of microRNAs in several types of cancers, the functional contributions of microRNAs to pancreatic cancer remain unclear.

Methods

In the present study, the expression levels of specific microRNAs identified by microarray analyses were examined in a panel of 15 pancreatic cancer cell lines. We then investigated the functional roles of these microRNAs in the proliferation and invasion of pancreatic cancer cells.

Results

Based on the microarray data, we found frequent and marked overexpression of miR-10a, miR-92, and miR-17-5p in pancreatic cancer cell lines. Microdissection analyses revealed that miR-10a was overexpressed in pancreatic cancer cells isolated from a subset of primary tumors (12 of 20, 60%) compared with precursor lesions and normal ducts (P?miR-10a inhibitors decreased the invasiveness of pancreatic cancer cells (P?HOXA1, a target of miR-10a, promoted the invasiveness of pancreatic cancer cells (P?Conclusions The present data suggest that miR-10a is overexpressed in a subset of pancreatic cancers and is involved in the invasive potential of pancreatic cancer cells partially via suppression of HOXA1.     

Inhibition of VEGFR-2 Reverses Type 1 Diabetes in NOD Mice by Abrogating Insulitis and Restoring Islet Function

The dysregulation of receptor tyrosine kinases (RTKs) in multiple cell types during chronic inflammation is indicative of their pathogenic role in autoimmune diseases. Among the many RTKs, vascular endothelial growth factor receptor (VEGFR) stands out for its multiple effects on immunity, vascularization, and cell migration. Herein, we examined whether VEGFR participated in the pathogenesis of type 1 diabetes (T1D) in nonobese diabetic (NOD) mice. We found that RTK inhibitors (RTKIs) and VEGF or VEGFR-2 antibodies reversed diabetes when administered at the onset of hyperglycemia. Increased VEGF expression promoted islet vascular remodeling in NOD mice, and inhibition of VEGFR activity with RTKIs abrogated the increase in islet vascularity, impairing T-cell migration into the islet and improving glucose control. Metabolic studies confirmed that RTKIs worked by preserving islet function, as treated mice had improved glucose tolerance without affecting insulin sensitivity. Finally, examination of human pancreata from patients with T1D revealed that VEGFR-2 was confined to the islet vascularity, which was increased in inflamed islets. Collectively, this work reveals a previously unappreciated role for VEGFR-2 signaling in the pathogenesis of T1D by controlling T-cell accessibility to the pancreatic islets and highlights a novel application of VEGFR-2 antagonists for the therapeutic treatment of T1D.In type 1 diabetes (T1D), genetic and environmental risk factors lead to immune dysregulation, provoking an autoimmune response directed toward insulin-producing β-cells of the islets of Langerhans. Previous investigations have estimated that β-cells or islets in nonobese diabetic (NOD) mice and humans are diminished to 10–30% of their initial mass (1,2), and the residual islets are largely dysfunctional when hyperglycemia is first detected (1,2). However, low levels of C-peptide can be detected in T1D patients as far out as 1–2 years postdiagnosis, indicating a window of opportunity for therapies that can restore or preserve islet mass and function (3).Multitarget receptor tyrosine kinase inhibitors (RTKIs), such as sunitinib, were originally designed to target malignant tumors that express dysregulated tyrosine kinases, including platelet-derived growth factor (PDGF)-R, c-FMS, or c-Kit. However, these inhibitors also target vascular endothelial growth factor (VEGF) receptors (VEGFRs), which are elevated in the parenchyma and tissue vasculature in many tumor microenvironments and during chronic inflammation. VEGF regulates vasculogenesis and angiogenesis largely through activation of VEGFR-2 (4). In addition to stimulating endothelial cell mitogenesis and cell migration, VEGF also has effects on a limited number of other cell types, including stimulation of monocyte/macrophage migration. Studies of transgenic mice lacking VEGFR-1 (5) or that express VEGFR-1 with a “dead” kinase domain (6) reveal that VEGFR-1 functions as a negative regulator of vasculogenesis and angiogenesis. Similarly, VEGFR-2 deficiency is embryonically lethal in mice but is attributed to a nonfunctional and underdeveloped vascular system (7). The phenotypes of VEGFR-1 and VEGFR-2–null mice indicate that, although VEGF-A has limited function through VEGFR-1, the vascular remodeling functions of VEGF-A are largely mediated through the activation of VEGFR-2.Tyrosine kinase inhibitors (TKIs) have shown efficacy in mouse models of muscular dystrophy (8), multiple sclerosis (9), rheumatoid arthritis (10–12), and psoriasis (13). TKI can prevent and reverse diabetes in NOD mice (14–16). Imatinib, which predominantly targets c-abl and PDGF, reversed diabetes in NOD mice (14), but other RTKIs with distinct inhibitory profiles (e.g., sunitinib) were even more effective, suggesting that the precise constellations of TK targets were critical for maximum efficacy. In this regard, the VEGF-A/VEGFR-2 pathway, a key target of sunitinib, stands out as a key kinase regulating the pathogenesis of several of these inflammatory disorders (17–19). Intriguingly, VEGF serum levels are elevated in T1D patients compared with healthy controls and positively correlate with increased HbA_1c levels (20).In this study, we determined whether VEGFR-2 might be involved in the pathogenesis of T1D and tested the therapeutic efficacy of VEGFR-2 inhibition in the NOD mouse model of T1D. We report that inhibition of VEGFR-2 by RTKIs or blocking antibodies rapidly reversed diabetes and maintains euglycemia with continued drug administration. Reversal of diabetes was attributed to an abrogation of vascular remodeling in the pancreatic islets, which impairs T-cell trafficking and the severity of insulitis, ultimately improving glucose tolerance. Histological analysis of human and mouse pancreata revealed a positive correlation between the severity of insulitis and islet vascularity, implicating inflammation as a major driving force in the vascular remodeling observed in the islets. Collectively, our findings suggest that VEGF/VEGFR-2 signaling serves a critical gatekeeper function by controlling essential remodeling of the vasculature that is necessary for T cells to gain access to tissues.     

Pim-3 promotes the growth of human pancreatic cancer in the orthotopic nude mouse model through vascular endothelium growth factor

Background

As one of the most lethal cancers, pancreatic cancer presents poor prognosis with an overall 5-y survival of less than 5%. We previously reported that Pim-3, a member of the proto-oncogene Pim family that encodes serine/threonine kinases, is aberrantly expressed in human pancreatic cancer lesions. In the current study, we investigated the role of Pim-3 in promoting tumor growth and angiogenesis in an orthotopic nude mouse model of human pancreatic cancer.

Methods

We constructed retroviral vectors for human Pim-3 and a kinase-dead mutant of human Pim-3 (K69M); the retroviral supernatants generated from these vectors were then used to infect the human pancreatic cancer cell line MiaPaCa-2 to establish stable cell lines. We assessed cell proliferation using CCK-8, tumor growth, and angiogenesis in vivo in an orthotopic mouse model of pancreatic cancer. While tumor size was measured using magnetic resonance imaging, the tumor tissues were excised for protein extraction and histological analysis to detect vascular endothelium growth factor (VEGF) expression and vessel density.

Results

We established an orthotopic nude mouse model of human pancreatic cancer. We observed that Pim-3 promoted the proliferation of human pancreatic cancer cells, both in vitro and in vivo. Moreover, Pim-3 is required for vasculogenesis of primary human pancreatic tumors in vivo and promotion of angiogenesis through the induction of VEGF expression.

Conclusions

Pim-3 can promote tumor growth and angiogenesis by stimulating the VEGF pathway.     

Osteopontin Expression is Associated with Improved Survival in Patients with Pancreatic Adenocarcinoma

Background

Osteopontin (OPN) is a secreted protein of the extracellular matrix. It has been used as a marker for tumor aggressiveness and correlated with clinical outcomes in several solid tumors, such as liver, lung, and breast. We determined the OPN expression and its influence on survival in patients with resected pancreatic adenocarcinoma.

Methods

Tissue microarrays were constructed from 245 resected pancreatic adenocarcinomas. Immunohistochemical staining for OPN was undertaken and compared to normal pancreas (n?=?12). OPN expression was then correlated with patient demographics, tumor size, grade, node, and margin status. Survival curves were created by the Kaplan?CMeier method and compared by log rank analysis.

Results

In total, 181 (74?%) of pancreatic adenocarcinoma tissues expressed OPN compared to 7 (58?%) of normal controls (p?=?0.004). Expression was observed predominantly in the cytoplasm of the tumor cells. The median and 2?year overall survival was longer when OPN was expressed (17.1 vs. 11.6?months, and 38 vs. 24?%, respectively, p?=?0.04). Multivariate analysis showed OPN expression and T stage to be independent predictors of overall survival, while other histopathologic factors such as tumor grade, tumor size, and nodal status were not.

Conclusions

These results suggest that the presence of OPN expression in pancreatic adenocarcinoma may have a protective effect independent of tumor stage. This emphasizes the importance of the interaction between pancreatic cancer cells and their stromal elements.     

Genomic Analysis of Pterostilbene Predicts Its Antiproliferative Effects Against Pancreatic Cancer In Vitro and In Vivo

Background

To investigate the inhibitory role of pterostilbene in pancreatic cancer, we conducted a genomic analysis of pterostilbene-treated pancreatic cancer cells. We also investigated the effect of pterostilbene upon the carcinogenic markers, manganese superoxide dismutase, cytochrome C, Smac/DIABLO, and STAT3 phosphorylation in vitro. The antiproliferative effects of pterostilbene were further evaluated in an in vivo model.

Methods

Pancreatic cancer cells were treated with pterostilbene and evaluated with DNA microarray analysis. Pterostilbene-treated cells were analyzed for cytochrome C, Smac/DIABLO, manganese superoxide dismutase (MnSOD)/antioxidant activity, and STAT3 phosphorylation using ELISA. Data were statistically analyzed using ANOVA. Pterostilbene was then administered to nude mice for 8?weeks, and tumor growth rates were recorded and statistically analyzed.

Results

Microarray analysis of pterostilbene-treated cells revealed upregulation of pro-apoptosis genes. In vitro, pterostilbene treatment altered levels of phosphorylated STAT3, MnSOD/antioxidant activity, cytochrome C, and Smac/DIABLO. In nude mice, oral pterostilbene inhibited tumor growth rates.

Conclusion

Pterostilbene alters gene expression in pancreatic cancer and increases the antiproliferative markers cytochrome C, Smac/DIABLO, and MnSOD/antioxidant activity. It was also shown to inhibit phosphorylated STAT3, a marker of accelerated tumorigenesis, and decrease pancreatic tumor growth in vivo. Further studies are warranted to elucidate the effects of pterostilbene in humans.     

Metastasenresektion nach neoadjuvanter Systemtherapie mit Multityrosinkinaseinhibitoren beim metastasierten Nierenzellkarzinom

Background

Metastatic renal cell carcinoma (mRCC) still poses a challenge to therapists in spite of the availability of multiple innovative molecular treatment options. Complete remission is rare and in cases of partial remission it is often unclear if necrosis or vital carcinoma tissue persists. We report on a cohort of patients who underwent metastasectomy after neoadjuvant therapy with multi-tyrosine kinase inhibitors (MTKI).

Methods

In 2009 a total of 11 patients (7 male and 4 female) underwent metastasectomy after achievement of ≥ 3 months stable partial remission. All patients received either sunitinib (n=7, mean 5.5 cycles), bevacizumab and interferon (IFN)-α2a (n=2, mean 8.5 months), temsirolimus (n=1, mean 9 months) or a combination of sunitinib followed by temsirolimus (n=1). Of the patients 7 presented with retroperitoneal lymph node metastases with a mean diameter of 3.5-12?cm, 2 patients with pulmonary metastases, 1 patient with lymph node and pancreas tail metastases and 1 female patient showed residual disease in the vena cava.

Results

All metastases were completely resected with negative surgical margins. In 82% of the cases histologically active, Ki-67 positive renal cell cancer tissue was identified. The following adjunctive interventions were necessary: vena cava resection with vascular prosthesis and reimplantation of the renal vein (n=3), partial liver resection (n=1), splenectomy (n=1) and pancreas tail resection (n=1). There were no significant perioperative complications but 1 patient developed fascial dehiscence and underwent revision surgery and 1 patient developed clinically insignificant pancreatitis. After a median follow-up of 12 months (range 8-19 months) 5 patients had no recurrence and 6 of the patients showed liver (n=3), lung (n=2) or bone (n=1) recurrences from which 3 patients died.

Conclusions

Metastasectomy of mRCC is associated with a low rate of complications in experienced centers. Surgical resection of metastatic disease is indicated to achieve complete remission with a favorable prognosis because of biologically active kidney cancer tissue. Patients with isolated and resectable metastases are ideal candidates for such a procedure.     

Perioperative Complications After Neoadjuvant Chemotherapy With and Without Bevacizumab for Colorectal Liver Metastases

Purpose

Bevacizumab has been shown to increase progression free and overall survival in patients with metastatic colorectal cancer. Neoadjuvant bevacizumab is commonly used in patients undergoing liver resection. Our purpose was to evaluate whether bevacizumab is associated with increased rate of perioperative complications in patients undergoing hepatic resection for colorectal liver metastases (CRLM).

Methods

Retrospective analysis of patients undergoing hepatic resection for CRLM who received chemotherapy and bevacizumab (group 1, n?=?134), or chemotherapy alone (group 2, n?=?57). We compared demographics, surgical characteristics, and perioperative course.

Results

Perioperative complications developed in 35 % of patients in group 1, and 47 % in group 2 (p?=?0.11). Of those complications, 15 (11.2 %) in group 1, and 5 (8.8 %) in group 2 were considered major (p?=?0.617). Four patients, all of whom received preoperative bevacizumab, developed enteric leaks following combined liver and bowel resection. The rate of anastomotic leak in group 1 was 10 %, compared with 0 in group 2, p?=?0.56.

Conclusion

Neoadjuvant chemotherapy along with bevacizumab was not associated with an increased risk of postoperative complications after hepatic resection. Possible association of increased morbidity with simultaneous bowel and liver resections following bevacizumab administration was found and we recommend avoiding such treatment combination.     

Hemoglobin-Based Oxygen Carrier Mitigates Transfusion-Mediated Pancreas Cancer Progression

Background

Perioperative blood transfusion in pancreatic cancer patients is linked to decreased survival; however, a causal mechanism has not been determined. Previously we have shown that the plasma fraction of stored packed red blood cells (pRBCs) promotes pancreas cancer progression and associated morbidity. We hypothesize these untoward effects will be mitigated by use of a hemoglobin-based oxygen carrier (HBOC).

Methods

Cytokines and growth factors were measured in the plasma fraction from stored pRBCs and in an HBOC via cytokine array followed by formal enzyme-linked immunosorbent assay (ELISA). In an immunocompetent murine model, pancreas cancer progression was determined in vivo by bioluminescence, tumor weight, and number of metastases.

Results

Elevated levels of epidermal growth factor (EGF), platelet-derived growth factor BB (PDGF-BB), and regulated upon activation, normal T cell expressed and secreted (RANTES) were present in the plasma fraction of stored pRBCs, but were not found in the HBOC. Intravenous delivery of plasma fraction to mice with pancreatic cancer resulted in increased bioluminescence activity compared with mice that received HBOC. Metastatic events and pancreatic primary tumor weights were significantly higher in animals receiving plasma fraction from stored pRBCs compared with animals receiving HBOC.

Conclusions

Intravenous receipt of the acellular plasma fraction of stored pRBCs promotes pancreatic cancer progression in an immunocompetent mouse model. These untoward events are mitigated by use of an HBOC.     

Impact of Preoperative Bevacizumab on Complications After Resection of Colorectal Liver Metastases: Case-Matched Control Study

Background

Chemotherapy may increase postoperative morbidity and mortality after liver surgery. Especially bevacizumab, a monoclonal antibody against vascular endothelial growth factor (VEGF), could have a detrimental effect. To assess the impact of neoadjuvant bevacizumab on clinical outcome after hepatectomy for colorectal liver metastases (CRLMs) this case-matched control study was initiated.

Methods

The multicentric data collection was performed in the Swiss HPB Center of the University Hospital Zurich (CH), the Department of Digestive Surgery and Transplantation Strasbourg (F), and the Division of Hepato-biliary-pancreatic surgery of “Josep Tureta” Hospital Girona (E). Consecutive patients operated onbetween July 2005 and December 2007 due to CRLMs who received neoadjuvant chemotherapy were assessed. Patients were divided in two groups: group A had neoadjuvant chemotherapy with bevacicumab, and group B had it without bevacizumab.

Results

No differences in overall morbidity (56 vs. 40% in the bevacizumab and control groups, respectively, p = 0.23) or mortality could be documented. Similarly, the incidence of severe postoperative complications was not statistically different between the bevacizumab and control groups (31 and 18%, respectively, p = 0.31). Wound complications were comparable (11% in the bevacizumab group compared and 9% in the control group, p = 1.00). However, bevacizumab was associated with a significantly decreased incidence of postoperative hepatic insufficiency (7 vs. 20%, p = 0.03).

Conclusions

No impact on the incidence or severity of complications by bevacizumab could be shown. Bevacizumab may even reduce the incidence of liver failure after liver surgery.     

Patterns of Pancreatic Resection Differ Between Patients with Familial and Sporadic Pancreatic Cancer

Background

Although the increased risk of developing pancreatic cancer (PC) in families with a strong history of the disease is well known, characteristics and outcomes of patients with familial PC is not described well.

Aims

This study aims to evaluate outcomes following resection in patients with familial PC.

Methods

We studied 208 patients who underwent resection of PC from 2000 to 2007 and had prospectively completed family history questionnaires for the Biospecimen Resource for Pancreas Research at our institution. We compared clinical characteristics and outcomes of familial and sporadic PC patients.

Results

Familial (N?=?15) and sporadic PC patients (N?=?193) did not have significantly different demographics, pre-operative CA19-9, pre-operative weight loss, R0 status, or T-staging (all p??? 0.05). Familial PC patients had lower pre-operative total serum bilirubin concentrations (p?=?0.03) and lesions outside of the pancreatic head more frequently (p?=?0.02) than sporadic PC patients. There was no difference in survival at 2?years between familial and sporadic PC patients (p?=?0.52).

Conclusions

Familial PC patients appear to develop tumors outside of the pancreatic head more frequently than sporadic PC patients. This difference in tumor distribution may be due to a broader area of cancer susceptibility within the pancreas for familial PC patients.     

Predictive role of tissue-based molecular markers in patients treated with sunitinib for metastatic renal cell carcinoma

Introduction

We analyzed the clinicopathological data of patients undergoing radical nephrectomy for clear cell type renal cell carcinoma (RCC) who presented with metastasis and were subsequently treated with sunitinib and identified molecular markers in nephrectomy specimens to predict susceptibility to sunitinib.

Patients and methods

The medical records of 65 patients who underwent nephrectomy for metastatic clear cell type RCC and were then treated with sunitinib were reviewed. The nephrectomy specimens were subjected to prospective immunohistochemical staining for vascular endothelial growth factor, vascular endothelial growth factor receptor-2 (VEGFR-2), platelet-derived growth factor-B, and platelet-derived growth factor receptor-β expression.

Results

In 58 evaluable patients, the median value of initial and best overall response was ?9.69 and ?24.04 %, respectively. VEGFR-2 expression was associated significantly with initial and best overall responses to sunitinib, along with Karnofsky performance status and Memorial Sloan-Kettering Cancer Center prognostic risk group. Multiple linear regression analyses revealed that strong VEGFR-2 expression was positively associated with the best reduction in tumor response (β = ?0.275, P = 0.016) and poor Karnofsky performance status was negatively associated it (β = 0.477, P < 0.001). Karnofsky performance status and retroperitoneal lymph node involvement associated independently with progression-free- and overall survivals. None of the molecular markers associated significantly with survival.

Conclusion

VEGFR-2 expression might be a useful biomarker for predicting the response to sunitinib by patients with metastatic RCC. Karnofsky performance status and retroperitoneal lymph node involvement were predictive of disease progression and death.

     

Targeted therapy with bevacizumab and erlotinib tailored to the molecular profile of patients with recurrent glioblastoma. Preliminary experience

Background

Advances in comprehension of molecular biology of glioblastoma (GBM) have led to the development of targeted therapies. The aim of the present study was to evaluate the efficacy and safety of a targeted therapeutic approach in which administration of bevacizumab and erlotinib was tailored on the molecular profile of recurrent GBM.

Methods

We prospectively enrolled ten adult patients suffering from recurrent GBM who had undergone surgical resection and standard chemo-radiotherapy. Tumor tissue was assessed for the expression of EGFRvIII and MGMT promoter methylation by RT-PCR, and for PTEN and VEGF expression by immunohistochemistry. Normal PTEN status was required for inclusion. Patients with VEGF overexpressing tumors (10/10) were treated with bevacizumab (10 mg/kg iv every 2 weeks in 6-week?cycles); patients whose tumor expressed EGFRvIII (4/10) added erlotinib (150 mg/day orally; 300 mg/day if on enzyme-inducing antiepileptic drugs). Therapy was continued until disease progression or unacceptable toxicity. Primary endpoints of the study were response rate (RR), 6-month progression-free survival (PFS-6), and safety profile.

Results

The RR and PFS-6 were 100 % (4/4) and 50 % (3/6) in patients treated with bevacizumab+erlotinib (n?=?4) and bevacizumab (n?=?6), respectively. In the whole cohort (n?=?10), RR and PFS-6 were both 70 % (7/10); median PFS and overall survival (OS) were 8.0 (3.0–31.0) and 9.5 (5.0–31.0) months, respectively. No grade 3/4 adverse events were observed; three patients treated with bevacizumab+erlotinib displayed grade 1/2 rash not requiring dose reduction; one patient treated with bevacizumab developed intratumoral hemorrhage requiring treatment discontinuation.

Conclusion

To our knowledge, this is the first study on recurrent GBM in which administration of bevacizumab and erlotinib was tailored on the molecular profile of the patient’s tumor. Although we treated a limited number of patients, we obtained significantly higher RR and PFS-6 than those reported in a previous trial lacking molecular tumor analysis.     

Impact of microvessel density on lymph node metastasis and survival after curative resection of pancreatic cancer

Purpose  

The roles of angiogenesis and the most prominent angiogenic vascular endothelial growth factor (VEGF) in diseases of the pancreas remain controversial. We compared microvessel density (MVD) and VEGF status in normal pancreatic, chronic pancreatic, and pancreatic cancer (PC) tissues to establish their prognostic relevance.     

Dexamethasone stimulates expression of C-type Natriuretic Peptide in chondrocytes

Background

Endostatin is a C-terminal fragment of collagen XVIII which is a component of basement membranes with the structural properties of both collagens and proteoglycans. Endostatin has a major role in angiogenesis which is intimately associated with bone development and remodeling. Signaling between the endothelial cells and the bone cells, for example, may have a role in recruitment of osteoclastic precursor cells. Our study aims at exploring a possibility that endostatin, either as a part of basement membrane or as a soluble molecule, may control osteoclastogenesis and osteoclastic bone resorption in vitro.

Methods

Rat pit formation assay was employed in order to examine the effect of endostatin alone or in combination with vascular endothelial growth factor-A (VEGF-A) on bone resorption in vitro. Effect of these agents on osteoclast differentiation in vitro was also tested. Osteoclastogenesis and the number of osteoclasts were followed by tartrate resistant acid phosphatase (TRACP) staining and resorption was evaluated by measuring the area of excavated pits.

Results

Endostatin inhibited the VEGF-A stimulated osteoclastic bone resorption, whereas endostatin alone had no effect on the basal resorption level in the absence of VEGF-A. In addition, endostatin could inhibit osteoclast differentiation in vitro independent of VEGF-A.

Conclusion

Our in vitro data indicate that collagen XVIII/endostatin can suppress VEGF-A induced osteoclastic bone resorption to the basal level. Osteoclastogenesis is also inhibited by endostatin. The regulatory effect of endostatin, however, is not critical since endostatin alone does not modify the basal bone resorption.     

Pancreatic cancer in the remnant pancreas following primary pancreatic resection

Purpose

To clarify the clinical features of cancer in the pancreatic remnant.

Methods

We retrospectively reviewed the clinical and pathological findings of 10 patients who developed remnant pancreatic cancer in our hospital between 2002 and 2012. The KRAS sequences in both the initial pancreatic tumor and remnant pancreatic cancer were examined in two patients.

Results

Eight patients underwent a second pancreatectomy for remnant pancreatic cancer (resected group), while two patients were not operated on and underwent chemotherapy (unresected group). The remnant pancreatic cancer developed at the cut end of the pancreas (pancreaticogastrostomy site) in four patients. In the resected group, four patients died 17 months after the emergence of the remnant pancreatic cancer and four patients survived during the median 40.5-month observation period. The median survival of the unresected group after the emergence of the remnant pancreatic cancer was 10 months. The findings of the KRAS sequencing and immunohistological staining of the remnant pancreatic cancer for MUC1 and MUC2 in the two patients were consistent with those of the initial pancreatic tumor in one patient, and not consistent in the other.

Conclusions

Our results suggest that both local recurrence and a new primary cancer can develop in the pancreatic remnant, and repeated pancreatectomy can prolong survival.