Marker Of Tumor Aggressiveness Research Articles

68Ga-DOTATOC and FDG PET Imaging of Preclinical Neuroblastoma Models.

Somatostatine receptors subtype 2 (SSTR2) are regarded as a potential target in neuroblastoma (NB) for imaging and promising therapeutic approaches. The purpose of this study was to evaluate and compare the SSTR2 status by (68)Ga-[tetraxetan-D-Phe1, Tyr3]-octreotide ((68)Ga-DOTATOC) positron-emission tomography (PET) and the tumour metabolic activity by (18)F-fluorodeoxyglucose (FDG) PET in different experimental models of NB. Three cell lines of human NB with different levels of expression of SSTR2 were grafted into nude mice. Animals were imaged with FDG and (68)Ga-DOTATOC and the maximum standardized uptake value (SUVmax) was determined to quantify tracer uptake. Ex vivo biodistribution of (68)Ga-DOTATOC and immunohistochemical analysis of NB xenografts were performed. Compared with FDG, the SUVmax of (68)Ga-DOTATOC uptake by the tumour was lower but the ratio to background was higher; there was a strong positive correlation between SUVmax values observed with the two tracers (r(2)=0.65). Sorting the cell lines according to uptake of FDG or (68)Ga-DOTATOC, injected activity per gram of tissue, Ki67 index or expression of SSTR2 assessed visually led to the same classification. (68)Ga-DOTATOC allows preclinical imaging of NB according to the intensity of the expression of SSTR2. In contrast with what has been reported for neuroendocrine tumours, in this NB model, the (68)Ga-DOTATOC uptake was positively correlated with FDG uptake and with Ki67 index, usual markers of tumour aggressiveness. If confirmed in humans, this result would favour a theranostic application of (68)Ga-DOTATOC in NB, even in advanced stages.

Programmed death ligand 1 expression in hepatocellular carcinoma: Relationship With clinical and pathological features.

PD-L1 expression by either neoplastic or intratumoral inflammatory cells is related to tumor aggressiveness and suggests that the response to treatments targeting the PD-L1/PD-1 immune checkpoint could be restricted to particular HCC variants; thus, enrichment of these tumor subtypes in future clinical trials should be considered. (Hepatology 2016;64:2038-2046).

Quantitative apparent diffusion coefficient measurement obtained by 3.0 Tesla MRI as a potential noninvasive marker of tumor aggressiveness in breast cancer

PurposeTo assess the association between apparent diffusion coefficient (ADC), and histological prognostic parameters in malignant breast lesions. The ability of ADC to identify lesions with the presence of Lymphovascular invasion (LVI) in breast carcinoma was also examined. Materials and methodsThis HIPAA-compliant retrospective study consisted of 212 consecutive patients with known cancers who underwent 3.0T MRI between January 2011 and 2013. In this study, a total of 126 malignant lesions in 114 women, who had undergone DWI (b-values of 0 and 1000s/mm2) in addition to diagnostic MRI, were included. Patients with less than 0.8cm lesions, or those who underwent neoadjuvant chemotherapy or suboptimal DW images were excluded. Classical prognostic factors [lesion size, histopathological type and grade, lymph node (LN) status and lymphovascular invasion (LVI)], molecular prognostic markers [estrogen receptor (ER), progesterone receptor (PR) and human epidermal grow factor receptor 2 (HER2)] were reviewed and recorded. A region of interest (ROI) was drawn within the lesions to measure ADC values. Statistical analyses were performed by the Wilcoxon rank sum test (statistical significance at P<0.05). Adjusted p values from multiple comparison analysis were also calculated. ResultsThis study demonstrates an inverse correlation between ADC and LVI in malignant lesions and the ability of ADC to identify aggressiveness in lesions with positive LVI. Tumor size, grade, ER, PR, HER2 and lymph node status did not impact tumor ADC value. However, tumors with LVI showed significantly lower ADC values when compared to tumors without LVI, regardless of the enhancement type, histological grade, histological type, and LN status. ConclusionOur study shows that ADC could be a potential clinical adjunct in the evaluation of prognostic factors related to malignant lesion aggressiveness such as LVI.

Infection-related hospitalizations in breast cancer patients: Risk and impact on prognosis

Infections are a common cause of hospitalization in breast cancer patients. We studied the risk, clinical characteristics and outcomes of infection-related hospitalizations in this patient population. A Swedish registry-based study including 8338 breast cancer patients diagnosed between 2001 and 2008, followed prospectively for infection-related hospitalizations until 2010. Standardized incidence ratios (SIRs) were calculated using background rates from the general female population. Associations with clinical characteristics and mortality were analyzed using flexible parametric survival models. In total, 720 patients experienced an infection-related hospitalization during a median follow-up of 4.9 years. Infection rates were highest within the first year of diagnosis (SIR=5.61, 95% CI; 4.98-6.32), and site-specific risks were most pronounced for sepsis (SIR=3.14, 95% CI; 2.66-3.71) and skin infections (SIR=2.80, 95% CI; 2.24-3.50). Older age at diagnosis, comorbidities, markers of tumor aggressiveness, chemotherapy and axillary node dissection were independent predictors of infectious disease risk. Infection-related hospitalizations were also independently associated with overall and breast cancer-specific death. A significant number of breast cancer patients are hospitalized with an infection following diagnosis, which in turn predicts poor prognosis. The risk profile of infection-related hospitalizations is multifactorial, including patient, tumor and treatment-related factors.

Significance of Microscopically Incomplete Resection Margin After Esophagectomy for Esophageal Cancer.

The objectives of this study were to establish if R1 resection margin after esophagectomy was (i) a poor prognostic factor independent of patient and tumor characteristics, (ii) a marker of tumor aggressiveness and (iii) to look at the impact of adjuvant treatment in this subpopulation. Data were collected from 30 European centers from 2000 to 2010. Patients with an R1 resection margin (n = 242) were compared with those with an R0 margin (n = 2573) in terms of short- and long-term outcomes. Propensity score matching and multivariable analyses were used to compensate for differences in baseline characteristics. Independent factors significantly associated with an R1 resection margin included an upper third esophageal tumor location, preoperative malnutrition, and pathological stage III. There were significant differences between the groups in postoperative histology, with an increase in pathological stage III and TRG 4-5 in the R1 group. Total average lymph node harvests were similar between the groups; however, there was an increase in the number of positive lymph nodes seen in the R1 group. Propensity matched analysis confirmed that R1 resection margin was significantly associated with reduced overall survival and increased overall, locoregional, and mixed tumor recurrence. Similar observations were seen in the subgroup that received neoadjuvant chemoradiation. In R1 patients adjuvant therapy improved survival and reduced distant recurrence however failed to affect locoregional recurrence. This large multicenter European study provides evidence to support the notion that R1 resection margin is a prognostic indication of aggressive tumor biology with a poor long-term prognosis.

Abstract P1-07-22: Infectious disease hospitalization in breast cancer patients: Risk and impact on prognosis

Abstract Background: Infection-related hospitalizations are a serious complication in breast cancer patients, resulting in treatment delay, prolonged hospitalization and future morbidity and mortality. Little, however, is known about the actual risk, clinical characteristics and outcomes of infection-related hospitalizations in this patient population. Methods: We conducted a prospective population-based study including 7071 women diagnosed with primary invasive non-metastatic breast cancer between 2001 and 2008 in the Stockholm-Gotland region (Sweden), with complete follow-up until 2010. Standardized incidence ratios (SIRs) for infection-related hospitalizations (overall and by site) were estimated using background rates from the general female population, matched on age and calendar period. Associations with clinical characteristics and breast cancer outcomes (breast cancer death, distant metastasis and locoregional recurrence) were analysed using flexible parametric survival models. Results: During a medium follow-up of 5.3 years, 657 hospital admissions with infections were observed and 1,2 and 5-year cumulative risks were 3.4, 4.7 and 8.1% respectively. Rates of infection-related hospitalizations were increased compared to the general female population (SIR = 2.12; 95% CI = 1.96-2.29) and site-specific SIRs were most pronounced for sepsis (SIR = 3.65; 95% CI = 3.13-4.26) and skin infections (SIR = 2.93; 95% CI = 2.35-3.64). The overall risk of infections was highest during the first year (SIR = 5.06; 95% CI, 4.46-5.74), and recurrent disease contributed to the long-term risk observed. Older age at diagnosis, chemotherapy, axillary radiotherapy, comorbidities and markers of tumor aggressiveness (large tumors, estrogen receptor negative tumors and lymph node positive tumors) were independent predictors of infectious disease risk. Analyses evaluating the prognostic impact of infections revealed an independent effect on breast cancer death and distant metastasis, associations that were mainly driven by sepsis (HR = 4.53; 95% CI, 3.25-6.33 and 1.69; 95% CI, 1.04-2.79 respectively). No association was found with future risk of locoregional recurrence. Conclusions: Physicians and patients should be aware of the risk of serious infections which persists beyond the initial treatment period. Infection-related hospitalizations are an independent marker of poor prognosis. Further research is needed to elucidate the role of sepsis in breast cancer progression. Citation Format: Brand JS, Colzani E, Johansson A, Giesecke J, Clements M, Bergh J, Hall P, Czene K. Infectious disease hospitalization in breast cancer patients: Risk and impact on prognosis. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-07-22.

Enhanced expression of melanoma progression markers in mouse model of sleep apnea

IntroductionObstructive sleep apnea has been associated with higher cancer incidence and mortality. Increased melanoma aggressivity was reported in obstructive sleep apnea patients. Mice exposed to intermittent hypoxia (IH) mimicking sleep apnea show enhanced melanoma growth. Markers of melanoma progression have not been investigated in this model. ObjectiveThe present study examined whether IH affects markers of melanoma tumor progression. MethodsMice were exposed to isocapnic IH to a nadir of 8% oxygen fraction for 14 days. One million B16F10 melanoma cells were injected subcutaneously. Immunohistochemistry staining for Ki-67, PCNA, S100-beta, HMB-45, Melan-A, TGF-beta, Caspase-1, and HIF-1alpha were quantified using Photoshop. ResultsPercentage of positive area stained was higher in IH than sham IH group for Caspase-1, Ki-67, PCNA, and Melan-A. The greater expression of several markers of tumor aggressiveness, including markers of ribosomal RNA transcription (Ki-67) and of DNA synthesis (PCNA), in mice exposed to isocapnic IH than in controls provide molecular evidence for a apnea–cancer relationship. ConclusionsThese findings have potential repercussions in the understanding of differences in clinical course of tumors in obstructive sleep apnea patients. Further investigation is necessary to confirm mechanisms of these descriptive results.

Comparison of the influence of human epidermal growth factor receptor 2/neu on the markers of aggression in gastric adenocarcinomas and ductal carcinoma breast

Background: Human epidermal growth factor receptor 2/neu (HER2/neu) assessment is carried out in gastric adenocarcinomas (GAC) and invasive ductal carcinoma (IDC) breast to decide upon of trastuzumab targeted therapy. The study the only of its kind which has compared the influence of HER2/neu expression to markers of tumor aggression in these two malignancies in similar target population. Material and Methods: HER2/neu evaluation was conducted by immunohistochemistry (IHC) using the guidelines of Trastuzumab for Gastric Cancer trial for GAC and ASCO-CAP 2013 for IDC breast. The study correlated HER2/neu expression with markers of tumor aggression. Aims: The present retrospective study compares the disease influence of HER2/neu over expression on markers of disease aggression like age at presentation, grade of tumor and lymph node metastasis in GAC and IDC. Results: We have compared 82 cases of GAC with 113 cases of IDC. HER2/neu expression was seen in 23.2% and 36.3% of GACs and IDCs respectively. The mean age of HER2/neu positive GACs was found to be 3.47 years greater than their negative counterpart but the pattern was reversed in cases of IDC breast where the mean age of HER2/neu positive tumors was 6.09 years less than their negative counterpart. Though no correlation was found between grade of tumor and HER2neu status in GACs (P = 0.2930), however in IDC breast a significant correlation HER2neu positivity was found with higher grade (P = 0.0002). Similarly though we did not find any correlation of HER2neu status with nodal metastasis in GACs (P 0.0511) but in IDCs a significant correlation was found of nodal metastasis with HER2/neu positivity (P = 0.0001). Conclusion: We conclude that HER2/neu expression is more frequent in IDC breast and is significantly associated with markers of aggression in contrast to its influence on the same markers in GACs.

Minichromosome maintenance protein 7 as prognostic marker of tumor aggressiveness in pituitary adenoma patients.

Ki-67 labeling index (LI) is currently regarded as a useful prognostic marker of pituitary adenoma (PA) clinical behavior, although its relevance as a reliable clinical indicator is far from being universally accepted, since both validations and criticisms are found in the literature. Minichromosome maintenance 7 (MCM7), a cell-cycle regulator protein, has been recently proposed as a marker of tumor aggressiveness in tumors from many sites, including the CNS. Therefore, we evaluated MCM7, in comparison to Ki-67, as a potential marker of clinical outcome in PA. In this single-institution retrospective study, 97 patients with PA (23 ACTH, 12 GH, 29 PRL, 10 FSH/LH, and 23 non-secreting adenomas) were recruited and the prognostic value of both MCM7 and Ki-67 was evaluated by immunohistochemical techniques. In addition, p53 nuclear expression and mitotic index were also evaluated. Twenty-six of the 97 PA patients recurred during the follow-up period. Cox's regression analysis showed that high nuclear expression of MCM7 LI, unlike Ki-67 LI, was directly associated with a higher (7.7-fold) risk of recurrence/progression. Kaplan-Meier analysis of recurrence/progression-free survival curves revealed that patients with high MCM7 LI (≥15%) had a shorter recurrence/progression-free survival than those with low MCM7 LI (<15%). Moreover, among patients with invasive tumors, high MCM7 LI identified those with the highest risk of recurrence/progression. Data from this study suggest that MCM7 is a prognostic marker of clinical outcome in PA patients, more reliable and informative than Ki-67.

LGR5 is associated with tumor aggressiveness in papillary thyroid cancer.

Leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5) is a cancer stem cell marker and a down-stream target in Wnt/β-catenin signaling. In human papillary thyroid cancer (PTC), over activation of Wnt/β-catenin has been associated with tumor aggressiveness. Using established human cell lines (TPC-1, KTC-1, Nthy-ori-3-1), we report LGR5 and R-spondin (RSPO1-3) overexpression in PTC and manipulate LGR5 and Wnt/β-catenin signaling via both pharmacologic and genetic interventions. We test the association of LGR5 tumor expression with markers of PTC aggressiveness using a Discovery Cohort (n = 26 patients) and a Validation Cohort (n = 157 patients). Lastly, we explore the association between LGR5 and the BRAFV600E mutation (n = 33 patients). Our results reveal that LGR5 and its ligand, RSPO, are overexpressed in human PTC, whereby Wnt/β-catenin signaling regulates LGR5 expression and promotes cellular migration. In two separate cohorts of patients, LGR5 and RSPO2 were associated with markers of tumor aggressiveness including: lymph node metastases, vascular invasion, increased tumor size, aggressive histology, advanced AJCC TNM stage, microscopic extra thyroidal extension, capsular invasion, and macroscopic invasion. As a biomarker, LGR5 positivity predicts lymph node metastasis with 95.5% sensitivity (95% CI 88.8%-98.7%) and 61% specificity (95% CI: 48.4%-72.4%) and has a negative predictive value (NPV) of 91.3% (95% CI 79.2%-97.5%) for lymph node metastatic disease. In human PTC, LGR5 is also strongly associated with the BRAFV600E mutation (p = 0.005). We conclude that overexpression of LGR5 is associated with markers of tumor aggressiveness in human PTC. LGR5 may serve as a future potential biomarker for patient risk stratification and loco regional metastases in PTC.

Prognostic utility of diffusion-weighted MRI in oesophageal cancer: is apparent diffusion coefficient a potential marker of tumour aggressiveness?

To investigate the role of the apparent diffusion coefficient (ADC) as a potential prognostic biomarker in the evaluation of the aggressiveness of oesophageal cancer. Between November 2009 and December 2013, 43 patients with evidence of oesophageal or oesophago-gastric junction cancer were referred to our institution and prospectively entered in our database. The final study population consisted of 23 patients (18 men; 5 women; mean age, 64.62±10.91years) who underwent diffusion-weighted Magnetic Resonance before surgical intervention. Specifically, 14 were directly treated with surgery and 9 were addressed to chemo/radiotherapy beforehand. Two radiologists independently measured mean tumour ADC and inter-observer agreement (Spearman's and intraclass correlation coefficient [ICC]) was assessed. In the univariate analysis, overall survival curves related to pathological ADC, pT, pN, tumour location and histotype were fitted using the Kaplan-Meier method. Survival curves were then compared using the log-rank test. Inter-observer reproducibility was very good (Spearman's rho=0.95; ICC=0.94). At a total median follow-up of 19months (2-49months), 4 patients had died. The median follow-up was 18.50months (5-49months) for the surgery-only group (1/4 events, 25%) and 24months (2-34months) for the chemo/radiotherapy group (3/4 events, 75%). Survival time at 48months for the overall population was 59% (±0.11), while for the surgery-only group and the chemo/radiotherapy group was 90% (±0.09) and 61% (±0.34), respectively. In the univariate analysis, ADC values below or equal to 1.4×10(-3)mm(2)/s were associated with a negative prognosis both in the total population (P=0.016) and in the surgery-only group (P<0.001). Despite the biggest limitation of our study (i.e. the small study population), we were able to show that pathological ADC could be considered a prognostic factor for oesophageal cancer. DWI might be introduced into clinical practice as a promising and reliable technique in the diagnostic pathway of this tumour.

TGF Beta1 Expression Correlates with Survival and Tumor Aggressiveness of Prostate Cancer.

Although biopsy Gleason score and clinical stage can be used to inform treatment decisions for prostate cancer, identifying molecular markers of tumor aggressiveness could lead to a more tailored approaches to therapy. In the present study, we investigated the association of transforming growth factor (TGF)-β1 levels and various markers of tumor aggressiveness and explore some potential mechanisms underlying the associations. We used human and murine prostate cancer cell lines and their respective hormone resistance sub-lines, in vitro and in vivo to examine the changes in tumor aggressiveness, as well as the pathway responsible for these changes. Furthermore, 105 prostate cancer biopsy specimens were analyzed to correlate the level of TGF-β1 with the clinical characteristics of patients. Our data revealed that activated TGF-β1 signaling resulted in more aggressive tumor growth and augmented the epithelial-mesenchymal transition. Activated IL-6 signaling was associated with TGF-β1 levels and the aggressive tumor features noted in TGF-β1-positive prostate cancers in vitro and in vivo. Furthermore, the TGF-β1 levels significantly correlated with Tregs accumulation in vivo. The clinical data indicated that TGF-β1 immunoreactivity had a moderate positive correlation with IL-6 staining, advanced clinical stage, higher Gleason score, and pretreatment PSA in patients with prostate cancer. TGF-β1 levels are significantly associated with aggressive prostate features. In vitro and in vivo alternations of TGF-β1 expression impacts tumor invasiveness, tumor growth rate and recruitment of immunosuppressive Treg cells in the tumor microenvironment. TGF-β1 expression may represent a clinical useful biomarker to guide prostate cancer treatment decisions.

Circulating Thyroid-Stimulating Hormone Receptor Messenger Rna as a Marker of Tumor Aggressiveness in Patients with Papillary Thyroid Microcarcinoma

We have previously shown that thyroid-stimulating hormone receptor messenger RNA (TSHR mRNA) is detectable in the peripheral blood of patients with papillary thyroid microcarcinoma (PTmC). The aim of this study was to analyze the utility of TSHR mRNA status as a marker of tumor aggressiveness in patients with PTmC. Preoperative TSHR mRNA values were obtained in 152 patients who underwent thyroidectomy and were found to have PTmC on final pathology. Clinical parameters were analyzed from an institutional review board-approved database using χ(2) and t tests. Preoperatively, TSHR mRNA was detected in the peripheral blood in 46% of patients, which was less than that for macroscopic papillary thyroid carcinoma (PTC) (80%) but higher than for benign thyroid disease (18%) (P<.001). The focus of cancer was larger in the TSHR mRNA-positive group compared to the negative group (0.41 vs. 0.30 cm, respectively, P = .015). The prevalence of tall-cell variant was higher in the TSHR mRNA positive group. The rates of lymph node (LN) metastasis (16% vs. 10%), multifocality (46% vs. 49%), and extra-thyroidal extension (10% vs. 5%) were similar between the TSHR mRNA-positive and-negative groups, respectively. In patients 45 years or older, rate of LN metastasis was higher in those who were TSHR mRNA positive (10%) versus negative (2%) (P = .039). TSHR mRNA positivity predicted a higher likelihood of radioactive iodine treatment (36% vs. 17%, P = .009) postoperatively. This study shows that TSHR mRNA, which is a marker of circulating thyroid cancer cells, is detectable in about half of patients with PTmC. The positivity of this marker predicts a higher likelihood of LN involvement in patients with PTmC who are 45 years or older.

MPGES-1 in prostate cancer controls stemness and amplifies epidermal growth factor receptor-driven oncogenicity.

There is evidence that an inflammatory microenvironment is associated with the development and progression of prostate cancer (PCa), although the determinants of intrinsic inflammation in PCa cells are not completely understood. Here we investigated whether expression of intrinsic microsomal PGE synthase-1 (mPGES-1) enhanced aggressiveness of PCa cells and might be critical for epidermal growth factor receptor (EGFR)-mediated tumour progression. In PCa, overexpression of EGFR promotes metastatic invasion and correlates with a high Gleason score, while prostaglandin E2 (PGE2) has been reported to modulate oncogenic EGFR-driven oncogenicity. Immunohistochemical studies revealed that mPGES-1 in human prostate tissues is correlated with EGFR expression in advanced tumours. In DU145 and PC-3 cell lines expressing mPGES-1 (mPGES-1SC cells), we demonstrate that silencing or ‘knock down’ of mPGES-1 (mPGES-1KD) or pharmacological inhibition by MF63 strongly attenuates overall oncogenic drive. Indeed, mPGES-1SC cells express stem-cell-like features (high CD44, β1-integrin, Nanog and Oct4 and low CD24 and α6-integrin) as well as mesenchymal transition markers (high vimentin, high fibronectin, low E-cadherin). They also show increased capacity to survive irrespective of anchorage condition, and overexpress EGFR compared to mPGES-1KD cells. mPGES-1 expression correlates with increased in vivo tumour growth and metastasis. Although EGFR inhibition reduces mPGES-1SC and mPGES-1KD cell xenograft tumour growth, we show that mPGES-1/PGE2 signalling sensitizes tumour cells to EGFR inhibitors. We propose mPGES-1 as a possible new marker of tumour aggressiveness in PCa.

PTH-155 Significance of microscopically incomplete resection margin following oesophagectomy for oesophageal cancer

<h3>Introduction</h3> The aim of this study was to establish if R1 resection margin after oesophagectomy was (i) a poor prognostic factor independent of patient and tumour characteristics, (ii) a marker of tumour aggressiveness and (iii) to look at the impact of adjuvant treatment in this subpopulation. <h3>Method</h3> Data was collected from 30 European centres from 2000–2010. Patients with an R1 resection margin (n = 242) were compared to those with an R0 margin (n = 2573) in terms of short- and long-term outcomes. Propensity score matching and multivariable analyses were used to compensate for differences in baseline characteristics. <h3>Results</h3> Important factors significantly associated with an R1 resection margin included an upper third oesophageal tumour location, preoperative malnutrition and pathological stage III. There were significant differences between the groups in postoperative histology, with an increase in pathological stage III and TRG 4/5 in the R1 group. Total average lymph node harvests were similar between the groups, however there was an increase in the number of positive lymph nodes seen in the R1 group. Propensity matched analysis confirmed R1 resection margin was significantly associated with reduced overall survival and increased overall, loco-regional, and mixed tumour recurrence. Similar findings were seen in the subgroup that received neoadjuvant chemoradiation. In R1 patients adjuvant therapy improved survival and reduced distant recurrence however failed to affect locoregional recurrence. <h3>Conclusion</h3> This large multi-centre European study provides evidence to support the notion that R1 resection margin is a prognostic indication of aggressive tumour biology with a poor long-term prognosis. <h3>Disclosure of interest</h3> None Declared.

(1) H NMR spectroscopy of glioblastoma stem-like cells identifies alpha-aminoadipate as a marker of tumor aggressiveness.

Patients suffering from glioblastoma multiforme (GBM) face a poor prognosis with median survival of about 14 months. High recurrence rate and failure of conventional treatments are attributed to the presence of GBM cells with stem-like properties (GSCs). Metabolite profiles of 42 GSC lines established from the tumor tissue of adult GBM patients were screened with (1) H NMR spectroscopy and compared with human neural progenitor cells from human adult olfactory bulb (OB-NPCs) and from the developing human brain (HNPCs). A first subset (n=12) of GSCs exhibited a dramatic accumulation of the metabolite α-aminoadipate (αAAD), product of the oxidation of α-aminoadipic semialdehyde catalyzed by the ALDH7A1 aldehyde dehydrogenase (ALDH) family in lysine catabolism. αAAD was low/not detectable in a second GSC subset (n=13) with the same neural metabolic profile as well as in a third GSC subset (n=17) characterized by intense lipid signals. Likewise, αAAD was not detected in the spectra of OB-NPCs or HNPCs. Inhibition of mitochondrial ATP synthase by oligomycin treatment revealed that the lysine degradative pathway leading to αAAD formation proceeds through saccharopine, as usually observed in developing brain. Survival curves indicated that high αAAD levels in GSCs significantly correlated with poor patient survival, similarly to prostate and non-small-cell-lung cancers, where activity of ALDH7A1 correlates with tumor aggressiveness.

Aerobic Glycolysis as a Marker of Tumor Aggressiveness: Preliminary Data in High Grade Human Brain Tumors.

Objectives. Glucose metabolism outside of oxidative phosphorylation, or aerobic glycolysis (AG), is a hallmark of active cancer cells that is not directly measured with standard 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET). In this study, we characterized tumor regions with elevated AG defined based on PET measurements of glucose and oxygen metabolism. Methods. Fourteen individuals with high-grade brain tumors underwent structural MR scans and PET measurements of cerebral blood flow (CBF), oxygen (CMRO2) and glucose (CMRGlu) metabolism, and AG, using 15O-labeled CO, O2 and H2O, and FDG, and were compared to a normative cohort of 20 age-matched individuals. Results. Elevated AG was observed in most high-grade brain tumors and it was associated with decreased CMRO2 and CBF, but not with significant changes in CMRGlu. Elevated AG was a dramatic and early sign of tumor growth associated with decreased survival. AG changes associated with tumor growth were differentiated from the effects of nonneoplastic processes such as epileptic seizures. Conclusions. Our findings demonstrate that high-grade brain tumors exhibit elevated AG as a marker of tumor growth and aggressiveness. AG may detect areas of active tumor growth that are not evident on conventional FDG PET.

Proteomic analysis of human glioblastoma cell lines differently resistant to a nitric oxide releasing agent.

Glioblastoma multiforme is the most aggressive astrocytoma characterized by the development of resistant cells to various cytotoxic stimuli. Nitric oxide (NO) is able to overcome tumor resistance in PTEN mutated rat C6 glioma cells due to its ability to inhibit cell growth by influencing the intracellular distribution of ceramide. The aim of this study is to monitor the effects of NO donor PAPANONOate on ceramide trafficking in human glioma cell lines, CCF-STTG1 (PTEN-mutated, p53-wt) and T98G (PTEN-harboring, p53-mutated), together with the assessment of their differential molecular signature by 2D-DIGE and MALDI mass spectrometry. In the CCF-STTG1 cell line, the results indicate that treatment with PAPANONOate decreased cell proliferation (<50%) and intracellular trafficking of ceramide, assessed by BODIPY-C5Cer, while these events were not observed in the T98G cell line. Proteomic results suggest that CCF-STTG1 cells are characterized by an increased expression of proteins involved in NO-associated ER stress (i.e. protein disulfide-isomerase A3, calreticulin, 78 kDa glucose-regulated protein), which could compromise ceramide delivery from ER to Golgi, leading to ceramide accumulation in ER and partial growth arrest. Conversely, T98G cell lines, resistant to NO exposure, are characterized by increased levels of cytosolic antioxidant proteins (i.e. glutathione-S-transferase P, peroxiredoxin 1), which might buffer intracellular NO. By providing differential ceramide distribution after NO exposure and differential protein expression of two high grade glioma cell lines, this study highlights specific proteins as possible markers for tumor aggressiveness. This study demonstrates that, in two different high grade glioma cell lines, NO exposure results in a different ceramide distribution and protein expression. Furthermore, this study highlights specific proteins as possible markers for tumor aggressiveness.

Clinical Utility of Altered Expressions of P53,Vascular Endothelial Growth Factor and Survivin in Patients with Bladder Cancer

Backgroundand Objectives: Bladder cancer is considered a major problem world wide. Aberrant P53, mutant vascular endothelial growth factor and deregulated expression of survivin were used as biochemical markers for investigation, prognosis and follow up. Aims: This study was conducted to assess the prognostic impact of altered expressions of these parameters in Erbil population with bladder cancer and correlated with other confounding factors age, gender and smoking effects, these factors also correlated with histopathologic characteristics such as grade and stage. Patients and Methods: This prospective study enrolled 50 newly diagnosed patients with bladder cancer, in addition 50 apparently healthy adults age – sex matched were also involved in this study. Serum parameters levels were measured using enzyme linked immunosorbent assay. Patients had these general criteria, newly discovered cases, no deep x-ray therapy, no chemotherapy, no hormonal therapy with histologic and cytologic confirmation of bladder cancer. Statistical Study: Data were analyzed using SPSS v. 18. Results: The mean serum P53,vascular endothelial growth factor and survivin levels in patient and control groups were P53= 1.682 ±0.665, 1.192 ± 0.284, vascular endothelial growth factor = 5.296 ±2.8, 2.000 ±0.704 and survivin = 5.468±0.715, 4.240±0.656 respectively. The statistical analysis revealed that, serum parameters levels were significantly increased in patients as compared with control group p< 0.001. Conclusion: Data revealed for the first time the relation between altered expressions of interested parameters in combination pattern with the incidence of bladder cancer in Erbil population, which has not previously reported in this region.This study tested the hypothesis and supported the concept that higher serum levels of these parameters might be a pathogenic and prognostic factors and a markers of tumor aggressiveness in bladder cancer. Early diagnosis is necessary for maximizing the rate of therapy. The gold standard care for the investigating of bladder cancer is cystoscopy which detects tumors accurately but it is invasive, expensive, and represents a high burden to the patients, and also, small papillary and flat-growing cancer may be missed. Regular cystoscopic examinations performed for monitoring the patients, because the recurrent rate is high. Therefore, this study was conducted to investigate the possibilities for replacing cystoscopy with more accurate, safe, no expensive, noninvasive diagnostic test and to recognize a new approach for providing opportunities for early detection which is an important goal to speed up the therapy of patients.

Gene and protein expression of glucose transporter 1 and glucose transporter 3 in human laryngeal cancer-the relationship with regulatory hypoxia-inducible factor-1α expression, tumor invasiveness, and patient prognosis.

Increased glucose uptake mediated by glucose transporters and reliance on glycolysis are common features of malignant cells. Hypoxia-inducible factor-1α supports the adaptation of hypoxic cells by inducing genes related to glucose metabolism. The contribution of glucose transporter (GLUT) and hypoxia-inducible factor-1α (HIF-1α) activity to tumor behavior and their prognostic value in head and neck cancers remains unclear. The aim of this study was to examine the predictive value of GLUT1, GLUT3, and HIF-1α messenger RNA (mRNA)/protein expression as markers of tumor aggressiveness and prognosis in laryngeal cancer. The level of hypoxia/metabolic marker genes was determined in 106 squamous cell laryngeal cancer (SCC) and 73 noncancerous matched mucosa (NCM) controls using quantitative real-time PCR. The related protein levels were analyzed by Western blot. Positive expression of SLC2A1, SLC2A3, and HIF-1α genes was noted in 83.9, 82.1, and 71.7 % of SCC specimens and in 34.4, 59.4, and 62.5 % of laryngeal cancer samples. Higher levels of mRNA/protein for GLUT1 and HIF-1α were noted in SCC compared to NCM (p < 0.05). SLC2A1 was found to have a positive relationship with grade, tumor front grading (TFG) score, and depth and mode of invasion (p < 0.05). SLC2A3 was related to grade and invasion type (p < 0.05). There were also relationships of HIF-1α with pTNM, TFG scale, invasion depth and mode, tumor recurrences, and overall survival (p < 0.05). In addition, more advanced tumors were found to be more likely to demonstrate positive expression of these proteins. In conclusion, the hypoxia/metabolic markers studied could be used as molecular markers of tumor invasiveness in laryngeal cancer.