Mast Cells Positive for c-Kit Receptor and Tryptase Correlate with Angiogenesis in Cancerous and Adjacent Normal Pancreatic Tissue

Background: Mast cells (MCs) contain proangiogenic factors, in particular tryptase, associated with increased angiogenesis in several tumours. With special reference to pancreatic cancer, few data have been published on the role of MCs in angiogenesis in both pancreatic ductal adenocarcinoma tissue (PDAT) and adjacent normal tissue (ANT). In this study, density of mast cells positive for c-Kit receptor (MCDP-c-KitR), density of mast cells positive for tryptase (MCDPT), area of mast cells positive for tryptase (MCAPT), and angiogenesis in terms of microvascular density (MVD) and endothelial area (EA) were evaluated in a total of 45 PDAT patients with stage T2–3N0–1M0. Results: For each analysed tissue parameter, the mean ± standard deviation was evaluated in both PDAT and ANT and differences were evaluated by Student’s t-test (p ranged from 0.001 to 0.005). Each analysed tissue parameter was then correlated to each other one by Pearson t-test analysis (p ranged from 0.01 to 0.03). No other correlation among MCDP-c-KitR, MCDPT, MCAPT, MVD, EA and the main clinical–pathological characteristics was found. Conclusions: Our results suggest that tissue parameters increased from ANT to PDAT and that mast cells are strongly associated with angiogenesis in PDAT. On this basis, the inhibition of MCs through tyrosine kinase inhibitors, such as masitinib, or inhibition of tryptase by gabexate mesylate may become potential novel antiangiogenetic approaches in pancreatic cancer therapy.

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Mast cells positive to c-kit receptor and to tryptase in normal to cancer pancreatic tissue and the correlation with angiogenesis.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.15_suppl.e16502 ◽

2020 ◽

Vol 38 (15_suppl) ◽

pp. e16502-e16502

Author(s):

Carmelo Laface ◽

Michele Ammendola ◽

Valeria Zuccalà ◽

Francesco Luposella ◽

Nicola Zizzo ◽

...

Keyword(s):

Standard Deviation ◽

Mast Cells ◽

Secretory Granules ◽

Mean Value ◽

T Test ◽

Ductal Adenocarcinoma ◽

Cancer Tissue ◽

Test Analysis ◽

Kit Receptor ◽

The Mean

e16502 Background: In the last years published studies demonstrated that mast cells (MCs) contain pro-angiogenic factors associated with tumoral angiogenesis. Tryptase is the most abundant and potent pro-angiogenic substance contained in MCs secretory granules and it can be released in tumour microenvironment. Up to now few data have been published on the role of MCs in angiogenesis in both pancreatic ductal adenocarcinoma cancer tissue (PDACT) and adjacent normal tissue (ANT). Methods: In this study MCs density positive to C-Kit receptor (MCDP-C-KitR), MCs density positive to tryptase (MCDPT), MCs area positive to tryptase (MCAPT), angiogenesis in terms of microvascular density (MVD) and endothelial area (EA) were evaluated by immunohistochemistry and image analysis in both PDACT and ANT. All tissue samples were obtained from 45 patients with stage T2-3N0-1M0 (by AJCC for Pancreas Cancer Staging 7th Edition) who had undergone surgery. For each analyzed tissue parameter mean ± standard deviation was evaluated in both PDACT and ANT and differences were evaluated by Student t-test. Each analysed tissue parameter was then correlated each other by Pearson t-test analysis. Results: The mean value ± standard deviation (SD) regarding MCDP-C-KitR, MCDPT, MCAPT, MVD and EA in TT was 14.69±4.57, 13.31± 4,23, 171.41±62,39 µ2, 29.11±7.93, 201.82±70.05 µ2 respectively and the mean value ± SD in ANT was 5.61 ± 2.39, 5.13 ± 2.03, 54.43 ± 16.73 µ2, 11.45 ± 4.96, 67.60 ± 21.96 µ2, respectively. Differences in terms of mean value ± SD between PDACT and ANT were significant for each analyzed tissue biomarker (p ranged from 0,001 to 0,004 by t-test analysis; Table). Data demonstrated that MCDP-C-KitR MCDPT, MCAPT, MVD and EA significantly increased from ANT to PCT. In PCT it was showed a correlation between MCDP-C-KitR and MCDPT (r = 0.87, p = 0.01), MCDP-C-KitR and MVD (r = 0.74, p = 0.02), MCDP-C-KitR and MCAPT (r = 0.81, p = 0.01), MCDPT and MVD (r = 0.72, p = 0.02), MCD-C-KitR and EA (r = 0.73, p = 0.02), MCDPT and MCAPT (r = 0.85, p = 0.01), MCAPT and MVD (r = 0.76, p = 0.02), MCAPT and EA (r = 0.66, p = 0.03), MCDPT and EA (r = 0.69, p = 0.03), MVD and EA (r = 0.82, p = 0.01). Conclusions: Our data suggest that assessed tissue parameters increased from ANT to PDACT and that MCs are associated with angiogenesis in PDACT. On this basis inhibition of MCs by tyrosine kinase inhibitors such as masitinib or inhibition of tryptase by gabexate mesylate may be a novel antiangiogenetic approach in pancreatic cancer therapy.

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CRM1 expression in pancreatic carcinoma.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.e15115 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. e15115-e15115 ◽

Cited By ~ 1

Author(s):

Amit Mahipal ◽

Domenico Coppola ◽

Shilpa Gupta ◽

Barbara Centeno ◽

Mokenge Peter Malafa

Keyword(s):

Pancreatic Cancer ◽

Pancreatic Carcinoma ◽

Anticancer Agents ◽

Normal Tissue ◽

Tissue Sample ◽

Pancreatic Tissue ◽

Ductal Adenocarcinoma ◽

Cancer Tissue ◽

Tissue Samples ◽

Paired Samples

e15115 Background: Increased expression of chromosomal region maintenance (CRM1) protein, also known as exportin 1, has been described in several human cancers. It is an important regulator of subcellular localization of tumor suppressor proteins and growth regulatory proteins. Direct inhibition of CRM1 blocks cell proliferation and induces apoptosis leading to the current evaluation of CRM1 inhibitors as anticancer agents in early phase clinical trials. There is a paucity of data regarding the prevalence of CRM1 expression in pancreatic cancer. Methods: We analyzed the expression of CRM1 by immunohistochemistry (IHC) in pancreatic tissue microarray (TMA) samples (malignant and non-malignant) obtained from patients who underwent potentially curative resection for pancreatic ductal adenocarcinoma. CRM1 antibody (Santa Cruz Biotechnology) was used for immunostaining the formalin fixed paraffin embedded core sections in the TMA samples. The intensity of staining and percentage of cells stained were graded on a scale of 0 to 3, with 3 being highest. The final IHC score was obtained using the product of immunostain intensity and percentage of cells stained (Range: 0-9). Low and high CRM1 expression was considered if the IHC score was 0 to 4 and 6 to 9 respectively. Results: Seventy nonmalignant and 91 pancreatic carcinoma samples were evaluated in this study. The median IHC score was 6 (range: 0-9) and 3 (range: 1-9) in malignant and nonmalignant pancreatic tissue samples respectively (P<0.0001). High CRM1 expression was found in 11% (8/70) of normal tissue samples and 69% (63/91) of tumor samples (P<0.0001). There were 48 paired samples of pancreatic cancer tissue and normal tissue obtained from same patient. Among these patients, 33 (69%) patients had higher CRM1 expression, 7 (15%) patients had similar expression and 8 (17%) patients had lower expression in malignant tissue sample as compared to their adjacent normal tissues. Conclusions: Higher CRM1 expression occurs frequently in pancreatic cancer as compared to nonmalignant pancreatic tissue, reinforcing its putative tumor oncogenic activity, and raising the value of targeting it for pancreatic cancer therapy.

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Effect of three-dimensional collagen I and global histone acetylation on gemcitabine resistance in pancreatic cancer cells.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.15_suppl.e14515 ◽

2012 ◽

Vol 30 (15_suppl) ◽

pp. e14515-e14515

Author(s):

Surabhi Dangi-Garimella ◽

Vaibhav Sahai ◽

Mario A. Shields ◽

Hidayatullah G. Munshi

Keyword(s):

Pancreatic Cancer ◽

Histone Acetylation ◽

High Mobility ◽

Tissue Microarrays ◽

Pancreatic Tissue ◽

Ductal Adenocarcinoma ◽

Pancreatic Tumors ◽

Collagen Gels ◽

Pancreatic Cancer Cells ◽

3D Collagen

e14515 Background: Pancreatic ductal adenocarcinoma (PDAC) is associated with a pronounced collagen-rich stromal reaction that has been shown to contribute to chemo-resistance. PDAC is also associated with epigenetic changes. We have previously shown that PDAC cells are resistant to gemcitabine in the collagen microenvironment because of increased expression of the chromatin remodeling protein high mobility group A2 (HMGA2) and increased ERK1/2 signaling. Methods: Pancreatic tissue microarrays were stained with trichrome and for histone H3K9, H3K27 acetylation (Ac), and histone acetyltransferase (HATs) expression. PDAC cells were plated onto tissue culture plastic or in 3D collagen gels and protein expression was assessed by Western blotting. DNA damage response was assessed by comet and clonogenic assays. Results: Human PDAC tumors display in areas of fibrosis higher histone H3K9Ac and H3K27Ac. Moreover, PDAC cells upregulate H3K9Ac and H3K27Ac along with GCN5, PCAF and p300 HATs when grown in 3D collagen. Inhibiting ERK1/2 activity and/or decreasing HMGA2 expression attenuates the effect of collagen on H3Ac and HAT expression. Human PDAC tumors with HMGA2 also demonstrate H3Ac and HAT expression. Additionally, cells in 3D collagen demonstrate reduced tailing with the comet assay, increased clonogenic potential and increased γH2AX following gemcitabine treatment, suggesting an increased repair response to damaged DNA in the collagen microenvironment. Significantly, downregulation of HATs along with inhibition of ERK1/2 activity attenuates gemcitabine-induced γH2AX detected in 3D collagen. Conclusions: Collagen microenvironment limits the effectiveness of gemcitabine through ERK1/2 and HMGA2-dependent HAT expression. HMGA2 expression is associated with histone acetylation and HAT expression in human PDAC tumors, particularly in area of fibrosis, suggesting that fibrosis may contribute to chemo-resistance through increased HMGA2-HAT signaling. Given that very little progress has been made in the treatment of pancreatic cancer, targeting HATs could be a novel approach to sensitize pancreatic tumors to chemotherapy.

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Candidate biomarkers identified in the saliva of patients with pancreatic adenocarcinoma.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.4_suppl.173 ◽

2012 ◽

Vol 30 (4_suppl) ◽

pp. 173-173

Author(s):

Jason B. Fleming ◽

Cynthia Trajtenberg

Keyword(s):

Mass Spectrometry ◽

Pancreatic Cancer ◽

Imaging Techniques ◽

Normal Subjects ◽

Ductal Adenocarcinoma ◽

P Value ◽

2D Dige ◽

Maldi Tof Mass Spectrometry ◽

Maldi Tof ◽

Student’S T

173 Background: There is currently no simple test to identify patients with pancreatic ductal adenocarcinoma (PDAC). The goal of this research is to develop such a method. We hypothesized that the salivary proteome of patients with localized PDAC contains a protein set distinct from normal subjects. Methods: We collected, processed and stored stimulated whole salivary gland secretions from patients with Stage I-II PDAC and aged-matched healthy volunteers. Fluorescently-labeled proteins were separated using 2D DIGE. Overlay imaging techniques compared 2D DIGE results, and spots representing individual proteins were quantified by signal, isolated from the gels, processed and protein identified using MALDI-TOF Mass Spectrometry. The SWISS-PROT database was used to identify proteins, and the ratio of signal in cancer versus normal samples was quantified and compared using Student’s t-test. Results: Samples from 10 PDAC patients and 10 healthy subjects were compared. A total of 96 spots on the 2D-DIGE comparison were uniquely present or absent in PDAC patient saliva. MALDI-TOF Mass Spectrometry examination of these 96 characterized 53 unique proteins. After eliminating duplicates and proteins without significant differences (p-value <0.02), a total of 30 proteins were identified with levels higher (n=18) or lower (n=12) in saliva from PDAC patients versus normal subjects. Conclusions: Examination of saliva from pancreatic cancer patients identified a set of proteins unique from normal subjects. These proteins represent targets for the development of biomarkers that could be used in early detection of pancreatic cancer.

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Tooth Size Discrepancies among Different Malocclusions in a Japanese Orthodontic Population

The Angle Orthodontist ◽

10.2319/101007-486.1 ◽

2008 ◽

Vol 78 (6) ◽

pp. 994-999 ◽

Cited By ~ 13

Author(s):

Toshiya Endo ◽

Ryota Abe ◽

Hiroo Kuroki ◽

Kenji Oka ◽

Shohachi Shimooka

Keyword(s):

Sex Differences ◽

T Test ◽

Class I ◽

Tooth Size ◽

Class Iii ◽

Test Analysis ◽

Size Correction ◽

Clinically Significant ◽

Overall Ratio ◽

Student’S T

Abstract Objective: To identify the possible sex differences in anterior and overall tooth size ratios and to evaluate whether any differences exist in tooth size ratios and distributions of subjects with clinically significant tooth size discrepancies among Angle Class I, Class II, and Class III malocclusion groups with the corresponding skeletal characteristics in a Japanese population. Materials and Methods: Each malocclusion group comprised 60 subjects (30 males and 30 females). The mesiodistal width from first molar to first molar was measured on each pretreatment cast to the nearest 0.01 mm using digital calipers, and the anterior and overall ratios were calculated. Student's t-test, Welch t-test, analysis of variance, and χ2-test were performed for statistical analysis. Results: No statistically significant sex differences were found in anterior or overall ratio in any group. No significant differences in anterior or overall ratios were found among the malocclusion groups. No significant differences were found between the distributions of subjects with clinically significant tooth size discrepancies, categorized by the Bolton standard deviation definition and by the actual amount of change calculated for tooth size correction in millimeters, among the malocclusion groups except for the mandibular correction for the overall ratio between Class I and Class III subjects. Conclusion: Bolton's values can be used with confidence for the typical Japanese orthodontic population. The use of the actual millimeters of correction for the tooth size ratios could help orthodontists avoid underestimating the prevalence of clinically significant tooth size discrepancies.

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SOX9 is a critical regulator of TSPAN8-mediated metastasis in pancreatic cancer

Oncogene ◽

10.1038/s41388-021-01864-9 ◽

2021 ◽

Author(s):

Junjian Li ◽

Xiaoliang Chen ◽

Liqun Zhu ◽

Zhenghong Lao ◽

Tianhao Zhou ◽

...

Keyword(s):

Pancreatic Cancer ◽

Molecular Mechanisms ◽

Kinase Inhibitors ◽

Tumor Stage ◽

Human Pancreatic Cancer ◽

Ductal Adenocarcinoma ◽

Endogenous Expression ◽

Egfr Tyrosine Kinase Inhibitors

AbstractPancreatic ductal adenocarcinoma (PDAC) is the deadliest cancer mainly owing to its proclivity to early metastasis and the lack of effective targeted therapeutic drugs. Hence, understanding the molecular mechanisms underlying early invasion and metastasis by PDAC is imperative for improving patient outcomes. The present study identified that upregulation of TSPAN8 expression in PDAC facilitates metastasis in vivo and in vitro. We found SOX9 as a key transcriptional regulator of TSPAN8 expression in response to EGF stimulation. SOX9 modulation was sufficient to positively regulate endogenous expression of TSPAN8, with concomitant in vitro phenotypic changes such as loss of cell–matrix adherence and increased invasion. Moreover, increased SOX9 and TSPAN8 levels were shown to correlate in human pancreatic cancer specimens and downregulated in vitro by EGFR tyrosine kinase inhibitors. High expression of SOX9 and TSPAN8 has been associated with tumor stage, poor prognosis and poor patient survival in PDAC. In conclusion, this study highlights the importance of the EGF-SOX9-TSPAN8 signaling cascade in the control of PDAC invasion and implies that TSPAN8 may be a promising novel therapeutic target for the treatment of PDAC.

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Detection of circulating hypoxia-regulated miR-210 in pancreatic adenocarcinoma patients

Journal of Clinical Oncology ◽

10.1200/jco.2009.27.15_suppl.4624 ◽

2009 ◽

Vol 27 (15_suppl) ◽

pp. 4624-4624

Author(s):

A. S. Ho ◽

X. Huang ◽

H. Cao ◽

A. C. Koong ◽

Q. T. Le

Keyword(s):

Pancreatic Cancer ◽

Cancer Patients ◽

Fold Increase ◽

Adverse Outcomes ◽

T Test ◽

Cellular Mechanisms ◽

C Elegans ◽

Novel Mirna ◽

Student’S T ◽

Student’S T Test

4624 Background: MicroRNAs (miRs) are small non-coding transcripts involved in many cellular mechanisms, including tumorigenesis. miR-210, in particular, has been shown to be induced by hypoxia, over-expressed in several different cancers, and correlated with adverse outcomes in breast cancer. Moreover, since pancreatic adenocarcinomas have been previously shown to be extremely hypoxic, we hypothesized that miR-210 may be elevated in the plasma of these patients compared to non-cancer controls. Here, we compared the circulating plasma levels of miR-210 in pancreatic cancer patients and controls using a novel miRNA extraction approach and quantitative PCR. Methods: Pretreatment EDTA plasma samples were obtained from pancreatic cancer patients and age-matched non-cancer controls. miRNA was extracted from 40ul of plasma and reverse transcribed to cDNA. A known quantity of c. elegans miR-54 was added to the sample as a normalization control. miR-210 and cel-miR-54 were then quantified using TaqMan MicroRNA Assays. The procedure was performed on the initial 11 pairs of age-matched pancreatic cancer patients and non- cancer controls, then validated with a second cohort of 12 pancreatic cancer patients and 11 controls. Results: miR-210 was reliably detected and quantified in small amounts of plasma using the approach developed in our study. There is a statistically significant four-fold increase of mir-210 expression in pancreatic cancer patients compared to normal controls (Student's t-test, p <0.0001). This difference was confirmed in the validation group (Student's t-test, p<0.05). Conclusions: Circulating miR-210 levels can be readily measured from a small quantity of plasma using a novel extraction method. Its expression is significantly higher in the blood of pancreatic cancer patients compared to controls and may potentially serve as a useful biomarker for pancreatic cancer diagnosis. No significant financial relationships to disclose.

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Pancreatic-Polypeptide in the Human Pancreas: Expression and Quantitative Variation During Development and in Ductal Adenocarcinoma

Acta Medica (Hradec Kralove Czech Republic) ◽

10.14712/18059694.2019.2 ◽

2003 ◽

Vol 46 (1) ◽

pp. 9-14 ◽

Cited By ~ 1

Author(s):

Demetrio Tamiolakis ◽

Constantine Simopoulos ◽

Athanasia Kotini ◽

Ioannis Venizelos ◽

Theodoros Jivannakis ◽

...

Keyword(s):

Pancreatic Cancer ◽

Pancreatic Polypeptide ◽

Pancreatic Tissue ◽

Ductal Adenocarcinoma ◽

Human Pancreas ◽

Neoplastic Tissue ◽

Mantle Zone ◽

Tissue Sections ◽

Ductal Pancreatic Adenocarcinoma ◽

Significant Difference

Aim: To determine the immunoreactivity of pancreatic-polypeptide (PP) during the development of the human fetal pancreas and ductal pancreatic adenocarcinoma, given that, PP positive cells were demonstrated either into its embryonic anlage or into pancreatic cancer. Methods: Tissue sections from 15 pancreatic fetal specimens, and equal number of ductal adenocarcinoma specimens, were assessed. Results: The density of positive cells in the primitive exocrine ductal epithelium and endocrine epithelium was significantly higher than the relevant density in the neoplastic pancreatic tissue of mixed (ductal – endocrine) and pure ductal type (p1=0.001, p2<0.0005, p3 =0.046 and p4<0.0005 respectively). The above values were estimated during the 10th to 12th week. There was no significant difference in the density of positive cells in the mantle zone of the islets from the 13th to the 24th week, and the neoplastic tissue of mixed (p5=0.11) and pure ductal type (p6=0.23). Conclusion: The immunostaining for PP identifies a subgroup of pancreatic ductal adenocarcinomas with a neuroendocrine component, initially considered as pure ductal tumors, and mixed ductal and neuroendocrine tumors. This pattern of expression in neoplasms recapitulates the normal pattern during the embryonal development of the organ, raising the question of therapeutic efficacy of PP and analogues as potential adjuvant treatment of pancreatic cancer.

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Screening and Validation of Independent Predictors of Poor Survival in Pancreatic Cancer

Pathology & Oncology Research ◽

10.3389/pore.2021.1609868 ◽

2021 ◽

Vol 27 ◽

Author(s):

Shui Liu ◽

Yan Cai ◽

E. Changyong ◽

Jiyao Sheng ◽

Xuewen Zhang

Keyword(s):

Pancreatic Cancer ◽

Cox Regression ◽

Clinical Stage ◽

T Test ◽

Functional Similarity ◽

Hub Genes ◽

Vital Status ◽

Student’S T ◽

One Way Anova ◽

Student’S T Test

Pancreatic cancer is a digestive system malignant tumor with high mortality and poor prognosis, but the mechanisms of progression remain unclear in pancreatic cancer. It’s necessary to identify the hub genes in pancreatic cancer and explore the novel potential predictors in the prognosis of pancreatic cancer. We downloaded two mRNA expression profiles from Gene Expression Omnibus and The Cancer Genome Atlas Pancreatic Cancer (TCGA-PAAD) datasets to screen the commonly differentially expressed genes in pancreatic cancer by limma package in R. Subsequently, measurement of the functional similarity among the 38 DEGs in common was performed to identify the hub genes using GOSemSim package. Then, survival analysis and Cox regression were applied to explore prognosis-related hub genes using the survival package. Statistics analysis by two-tailed Student’s t-test or one-way based on TCGA-PAAD datasets and qPCR detection in clinical samples were performed to explore the correlations between expression of hub genes in pancreatic cancer tissues and clinical parameters. Based on integrated analysis of TCGA and GEO datasets, we screened 38 DEGs in common, which were all up-regulated. The functional similarity results showed that 10 DEGs including TSPAN1, MSLN, C1orf116, PKP3, CEACAM6, BAIAP2L1, PPL, RAB25, ERBB3, and AP1M2 in the DEGs in common, which had the higher average functional similarity, were considered as the hub genes. Survival analysis results and Cox regression analysis showed that TSPAN1, CEACAM6, as well as ERBB3 were all associated with poor overall survival of PC. qPCR results showed that the expression levels of TSPAN1 and ERBB3 were significantly upregulated in the PC tissues. The statistical analysis results revealed that TSPAN1 expression correlated significantly with histologic grade, T stage, clinical stage, and vital status by two-tailed Student’s t-test or one-way ANOVA; ERBB3 expression correlated significantly with T stage, clinical stage, and vital status by two-tailed Student’s t-test or one-way ANOVA. We found that TSPAN1 and ERBB3 could be independent predictors of poor survival in pancreatic cancer.

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