Microvessel Density Expression Research Articles

Immunohistochemical evaluation of tumor hypoxia and angiogenesis: Pathological significance and prognostic role in head and neck squamous cell carcinomas

Objectives: Tumor hypoxia and angiogenesis have been implicated in therapeutic resistance of head and neck squamous cell carcinomas (HNSCCs). Immunohistochemical evaluation of hypoxia-inducible factor-1 alpha (HIF-1 α), a hypoxia transcription factor, and vascular endothelial growth factor (VEGF), a hypoxia-responsive pro-angiogenic factor, can be exploited for prognostication and guiding treatment intensification or de-escalation decisions in HNSCC patients. The purpose of the study is to evaluate the immunohistochemical expression patterns of HIF-1 α and VEGF and the microvessel density (MVD) for angiogenesis in HNSCC and assess their pathological significance and prognostic role. Materials and Methods: In this cross-sectional study, immunohistochemical expression of HIF-1 α, VEGF, and MVD through Cluster of Differentiation (CD31) was evaluated in paraffin-embedded tumor resection tissue of 44 patients with HNSCC. Associations among HIF-1 α, VEGF, and MVD with clinicopathological variables were assessed. Statistical Analysis: For assessment of association between HIF-1α, VEGF and MVD by CD 31 immunohistochemical markers and other clinicopathological variables Pearson’s chi-square test and Fisher’s exact tests were used. Analysis of survival was done using Kaplan-Meier statistics. Also, the univariate and multivariate analysis were performed using the Cox proportional hazard regression model for the calculation of hazard ratios. Results: Nuclear expression of HIF-1 α showed significant association with MVD (P = 0.007) and cytoplasmic expression of HIF-1 α with histologic grade (P = 0.03). Overexpression of HIF-1 α was more frequent in T3/T4 stage. In addition to cytoplasmic staining, VEGF showed a unique nuclear expression pattern in four cases of advanced disease with nodal metastasis. Logistic regression analysis showed tumors with nuclear overexpression of HIF-1 α to have increased MVD (P = 0.05), and tumors with higher MVD to have a presence of lymphovascular invasion (P = 0.014). Multivariate analysis showed HIF-1 α nuclear overexpression to be significantly associated with decreased survival of patients (P = 0.05). Conclusions: Immunohistochemical overexpression of HIF-1 α and MVD quantification can serve as cost-effective tools for prognostication and treatment modification of HNSCC patients in resourcelimited settings.

Application of CD34 expression combined with three-phase dynamic contrast-enhanced computed tomography scanning in preoperative staging of gastric cancer.

Accurate preoperative staging of gastric cancer (GC), a common malignant tumor worldwide, is critical for appropriate treatment plans and prognosis. Dynamic three-phase enhanced computed tomography (CT) scanning for preoperative staging of GC has limitations in evaluating tumor angiogenesis. CD34, a marker on vascular endothelial cell surfaces, is promising in evaluating tumor angiogenesis. We explored the value of their combination for preoperative staging of GC to improve the efficacy and prognosis of patients with GC. To explore the evaluation value of CD34 expression + dynamic three-phase enhanced CT scanning in preoperative staging of GC. Medical records of 106 patients with GC treated at the First People's Hospital of Lianyungang between February 2021 and January 2023 were retrospectively studied. All patients underwent three-phase dynamic contrast-enhanced CT scanning before surgery, and CD34 was detected in gastroscopic biopsy specimens. Using surgical and pathological results as the gold standard, the diagnostic results of three-phase dynamic contrast-enhanced CT scanning at different T and N stages were analyzed, and the expression of CD34-marked microvessel density (MVD) at different T and N stages was determined. The specificity and sensitivity of three-phase dynamic contrast-enhanced CT and CD34 in T and N staging were calculated; those of the combined diagnosis of the two were evaluated in parallel. Independent factors affecting lymph node metastasis were analyzed using multiple logistic regression. The accuracy of three-phase dynamic contrast-enhanced CT scanning in diagnosing stages T1, T2, T3 and T4 were 68.00%, 75.00%, 79.41%, and 73.68%, respectively, and for diagnosing stages N0, N1, N2, and N3 were 75.68%, 74.07%, 85.00%, and 77.27%, respectively. CD34-marked MVD expression increased with increasing T and N stages. Specificity and sensitivity of three-phase dynamic contrast-enhanced CT in T staging were 86.79% and 88.68%; for N staging, 89.06% and 92.86%; for CD34 in T staging, 64.15% and 88.68%; and for CD34 in N staging, 84.38% and 78.57%, respectively. Specificity and sensitivity of joint diagnosis in T staging were 55.68% and 98.72%, and N staging were 75.15% and 98.47%, respectively, with the area under the curve for diagnosis improving accordingly. According to multivariate analysis, a longer tumor diameter, higher pathological T stage, lower differentiation degree, and higher expression of CD34-marked MVD were independent risk factors for lymph node metastasis in patients with GC. With high accuracy in preoperatively determining the invasion depth and lymph node metastasis of GC, CD34 expression and three-phase dynamic contrast-enhanced CT can provide a reliable basis for surgical resection.

PET/MRI of hypoxia and vascular function in ER-positive breast cancer: correlations with immunohistochemistry

ObjectivesTo explore the relationship between indices of hypoxia and vascular function from 18F-fluoromisonidazole ([18F]-FMISO)-PET/MRI with immunohistochemical markers of hypoxia and vascularity in oestrogen receptor–positive (ER +) breast cancer.MethodsWomen aged > 18 years with biopsy-confirmed, treatment-naïve primary ER + breast cancer underwent [18F]-FMISO-PET/MRI prior to surgery. Parameters of vascular function were derived from DCE-MRI using the extended Tofts model, whilst hypoxia was assessed using the [18F]-FMISO influx rate constant, Ki. Histological tumour sections were stained with CD31, hypoxia-inducible factor (HIF)-1α, and carbonic anhydrase IX (CAIX). The number of tumour microvessels, median vessel diameter, and microvessel density (MVD) were obtained from CD31 immunohistochemistry. HIF-1α and CAIX expression were assessed using histoscores obtained by multiplying the percentage of positive cells stained by the staining intensity. Regression analysis was used to study associations between imaging and immunohistochemistry variables.ResultsOf the lesions examined, 14/22 (64%) were ductal cancers, grade 2 or 3 (19/22; 86%), with 17/22 (77%) HER2-negative. [18F]-FMISO Ki associated negatively with vessel diameter (p = 0.03), MVD (p = 0.02), and CAIX expression (p = 0.002), whilst no significant relationships were found between DCE-MRI pharmacokinetic parameters and immunohistochemical variables. HIF-1α did not significantly associate with any PET/MR imaging indices.ConclusionHypoxia measured by [18F]-FMISO-PET was associated with increased CAIX expression, low MVD, and smaller vessel diameters in ER + breast cancer, further corroborating the link between inadequate vascularity and hypoxia in ER + breast cancer.Key Points• Hypoxia, measured by [18F]-FMISO-PET, was associated with low microvessel density and small vessel diameters, corroborating the link between inadequate vascularity and hypoxia in ER + breast cancer.• Increased CAIX expression was associated with higher levels of hypoxia measured by [18F]-FMISO-PET.• Morphologic and functional abnormalities of the tumour microvasculature are the major determinants of hypoxia in cancers and support the previously reported perfusion-driven character of hypoxia in breast carcinomas.

Histopathological Evaluation of Angiogenic Markers in Non-Hodgkin's Lymphoma.

Background Angiogenesis plays a key role in the development, maintenance, and progression of tumor. The incidence of non-Hodgkin's lymphoma (NHL) is increasing from the past three decades. Materials and Methods The aim of the study is to evaluate microvessel density (MVD) using CD34 monoclonal antibody and vascular endothelial growth factor (VEGF) using monoclonal antibody that were studied in pretreatment paraffin-embedded tissue samples of 60 cases. Results MVD was found to be increased in parallel with increasing grade of tumor. B-NHL had a mean MVD of 79.5 ± 8.8 (no./mm 2 ), while T-NHL had a mean MVD of 183 ± 37.6 (no./mm 2 ). VEGF expression was seen in 42 cases (70%), 20 cases (33.3%) showed strong VEGF expression, and the remainder showed either weak (36.6%) or no (30%) staining. Strong VEGF expression is seen in 100% cases of T-NHL and 77.7% cases of B-NHL. Mean MVD and VEGF expression was found to be correlated significantly with the histological grade of NHL ( p = 0.001 and p = 0.000, respectively). Average microvessel counts were 53, 82.9, and 130.8 vessels (no./mm 2 ) for negative, weak, and strong VEGF staining, respectively. These differences were statistically significant ( p = 0.005 for strong vs. negative and p = 0.091 for strong vs. weak VEGF staining individually). Conclusion As the grade of tumor progresses, the angiogenic potential also advances which seems to depend on VEGF. The presence of higher MVD in high-grade lymphomas can be utilized for antiangiogenic drugs.

The Modeling Analysis and Effect of CHI3L1 and CD31-Marked Microvessel Density in the Occurrence and Development of Cervical Squamous Cell Carcinoma.

Background Chitinase-3-like protein 1 (CHI3Ll) has been identified as a novel tumor marker in several cancers. The objective of this study was to detect the expression of Chitinase-3-like protein 1 (CHI3L1) and CD31-labeled microvessel density (MVD) in cervical squamous cell carcinoma (CSCC) and to assess its prognostic impact. Methods Elivision™ plus immunohistochemical method was used to detect CHI3L1 expression and MVD in different cervical tissues. We analyzed the relationship between CHI3L1 and MVD in CSCC tissues and investigated the relationship between CHI3L1, MVD, and clinicopathological parameters. Univariate and multivariate survival analyses were performed to assess the impact on progression-free survival (PFS) and overall survival (OS). Results The positive expression rate of CHI3L1 protein in CSCC tissues (69.9%, 72/103) was significantly higher than that in high-grade cervical intraepithelial lesions (53.3%, 32/60), low-grade cervical intraepithelial lesions (25%, 15/60), and normal cervical tissues (16.7%, 10/60). MVD values ranged from 6 to 64 in CSCC, and no microvascular formation was observed in normal cervical tissues, high-grade intraepithelial lesions, or low-grade intraepithelial lesions. The high expression of CHI3L1 and MVD was significantly correlated with the invasion depth, differentiation degree, vascular invasion, and lymph node metastasis of CSCC (all P < 0.05). In CSCC, the expression of MVD in the CHI3L1 high-expression group (41.35 ± 9.056) was significantly higher than that in the CHI3L1 low-expression group (23.26 ± 11.000, P < 0.05). On univariate Kaplan–Meier analysis, FIGO stage, tumor diameter, lymph node metastasis, vascular invasion, CHI3L1, and MVD of CSCC were related to the prognosis of PFS and OS (all P < 0.05); however, CHI3L1 and MVD were not independent prognostic factors. Conclusion CHI3L1 may be involved in the progression of cervical cancer. Its high expression can promote neovascularization in the tumor microenvironment. CHI3L1 is a potential therapeutic target in the context of cervical cancer.

The Value of IOTA Simple Rules Combined With CEUS Scoring System in the Diagnosis of Benign and Malignant Ovarian Masses and Its Correlation With MVD and VEGF: A Preliminary Study.

To investigate the diagnostic value of International Ovarian Tumor Analysis (IOTA) simple rules combined with contrast-enhanced ultrasound (CEUS) scoring system in the differential diagnosis of ovarian tumors, and the correlations of the scoring system with microvessel density (MVD) and vascular endothelial growth factor (VEGF). One hundred eighty-nine patients with ovarian tumors were examined by routine ultrasound and CEUS. The enhanced characteristics of CEUS were observed, and the masses were classified by IOTA simple rules. To compare the diagnostic value of IOTA simple rules combined with CEUS scoring system and IOTA simple rules in the diagnosis of ovarian tumors. Immunohistochemistry was used to detect the expression of MVD and VEGF in postoperative tissue samples. The correlations between the new scoring system with MVD and VEGF were analyzed. The sensitivity (93.98%), specificity (94.34%), positive predictive value (92.86%), negative predictive value (95.24%), and accuracy (94.18%) of IOTA simple rules combined with CEUS scoring system in the diagnosis of ovarian tumors were higher than those of IOTA simple rules alone (all P < .05). The score system was significantly positively correlated with MVD and VEGF, and the r values were 0.77 and 0.63, respectively (P < .001). IOTA simple rules combined with CEUS scoring system was helpful to improve the accuracy of ultrasound diagnosis of ovarian tumors, which was significantly correlated with MVD and VEGF. It could provide important reference information for treatment scheme formulation and prognosis evaluation.

Mast Cell Counts and Microvessel Density Expressions in Hodgkin’s Lymphoma and Reactive Lymphadenopathy in Children

Mast Cell Counts and Microvessel Density Expressions in Hodgkin’s Lymphoma and Reactive Lymphadenopathy in Children

Dynamic contrast-enhanced MRI perfusion parameters are imaging biomarkers for angiogenesis in lung cancer.

Quantitative parameters of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) may have the potential to reflect angiogenesis and proliferation of pulmonary neoplasms. To verify whether DCE-MRI can identify pulmonary neoplasm property and evaluate the correlation of DCE-MRI perfusion parameters with microvessel density (MVD) and Ki-67 in lung cancer. This study enrolled 65 patients with one pulmonary neoplasm who underwent computed tomography-guided percutaneous lung biopsy with pathological diagnosis (43 malignant, 22 benign; mean age = 59.71 ± 11.72 years). All patients did DCE-MRI before biopsy. Quantitative MRI parameters including endothelial transfer constant (Ktrans), flux rate constant (Kep), and fractional extravascular extracellular space (EES) volume (Ve) were calculated by extended Tofts linear model. MVD was evaluated by CD34-expressing tumor vessels. Proliferation was assessed by Ki-67 staining. The correlations of parameters with MVD and Ki-67 expression were analyzed. Ktrans and Kep values were significantly increased in malignant lesions compared to benign lesions (P = 0.001 and 0.022, respectively), whereas no statistical difference in Ve was found. The CD34 expression was positively correlated to Ktrans (r = 0.608; P = 0.004) and Kep (r = 0.556; P = 0.001). Subsequent subtype analyses also showed positive correlations of Ktrans and Kep with MVD in adenocarcinoma group (r = 0.550 and 0.563; P = 0.012 and 0.015, respectively). No significant correlation was found between these parameters and Ki-67. Ktrans and Kep may distinguish benign and malignant pulmonary neoplasm. Ktrans and Kep, with their positive correlation to MVD, can be used as non-invasive parameters reflecting lung cancer angiogenesis.

Clinical significance of CD34, podoplanin and Ki-67 expression in patients with locally advanced squamous cell cervical carcinoma

The aim of the study was to assess immunohistochemical CD34, podoplanin and Ki-67 expression in cervical tumour of patients with cervical squamous cell carcinoma (SCC) staged IIB and IIIB, a relationship with selected clinical and histological parameters and its prognostic significance. This prospective study included 52 patients. Microvessel density (MVD) by CD34, lymphatic vessel density (LVD) by podoplanin and the Ki-67 index in specimens from paraffin blocks with cervical SCC tissues were examined. The relationship between these data and selected clinical and histological parameters was analysed. Positive correlation of MVD and the Ki-67 index was observed. No correlation was observed for MVD, LVD and the Ki-67 index in the tumour with staging, grading, length of treatment and squamous cell carcinoma antigen (SCC-Ag) concentration before and after treatment. The expression of MVD, LVD and the Ki-67 index in cervical SCC did not contribute to the risk of relapse and cancer-related death. No relationship was found for MVD, LVD and the Ki-67 index in cervical tumour of patients with locally advanced cervical SCC with staging, grading and serum SCC-Ag level. MVD, LVD and the Ki-67 index in the tumour did not contribute to the risk of relapse or cervical SCC-related death. Impact Statement What is already known on this subject? In many patients, invasive cervical squamous cell carcinoma (SCC) is diagnosed in a locally advanced stage, when the prognosis depends on many well-known factors connected with tumour biology, staging and general condition of the patient. Despite numerous studies, the value of immunohistochemical CD34, podoplanin and Ki-67 expression in cervical tumour of these patients is still not well defined. What do the results of this study add? In our prospective study, no relationship for microvessel density (MVD), lymphatic vessel density (LVD) and the Ki-67 index in cervical tumour of patients with locally advanced cervical SCC with staging, grading and serum squamous cell carcinoma antigen (SCC-Ag) level was found. Additionally, MVD, LVD and the Ki-67 index in the tumour did not contribute to the risk of relapse or cervical SCC-related death. What are the implications of these findings for clinical practice and/or further research? Our study underlines the limited value of immunohistochemical CD34, podoplanin and Ki-67 expression in cervical tumour of patients with locally advanced cervical SCC. Further research should be focussed on identifying and validating novel prognostic and predictive factors.

Stromal microenvironment namely angiogenesis, tumor-infiltrating lymphocytes, and matrix metalloproteinase in invasive breast carcinoma: Do they have a prognostic role?

The role of stromal microenvironment in growth, invasiveness, and metastatic potential of breast carcinoma (BC) is being recognized increasingly, both to predict prognosis and as potential therapeutic targets. The present study aimed to evaluate the correlation of angiogenesis, tumor-associated lymphocytes, and stromal CD10 expression with clinicopathologic parameters. This study included 100 consecutive cases of invasive BC undergoing modified radical mastectomy. Relevant clinical details, pathological grade, lymph nodal status, and clinical stage were noted. Paraffin-embedded sections were subjected to immunohistochemistry for CD34, CD20, CD45RO, and CD10. Microvessel density (MVD), tumor-associated lymphocytes, and stromal CD10 expression were estimated from these sections. Statistical analysis was done using nonparametric tests to correlate the clinic-pathologic features with each of these parameters. MVD was found to be significantly higher in Grade III, node-positive cases, and higher stage breast cancers (P < 0.05). The number of T-lymphocytes was higher in node-positive cases, while B-lymphocytes were lower in number in higher grade tumors. CD10 expression showed a significant positive association with tumor grade, nodal status, and stage (P < 0.05 for each). This study demonstrates that changes in stromal microenvironment of BC such as MVD, tumor-associated lymphocytes, and stromal CD10 expression correlate with the clinicopathological parameters and hence may be exploited as prognostic markers or therapeutic targets, based on further larger studies.

Angiogenesis and osteopontin expression in paraskeletal myeloma with plasmablastic morphology and aggressive clinical course: a report of two cases

Extramedullary disease (EMD) of multiple myeloma (MM) can present as paraskeletal (paraosseous) plasmocytoma (PP) that arise from skeletal focal lesions or extramedullary plasmacytomas (EMP) that derive from hematogenous spread. The pathogenetic mechanisms that distinguish classical MM, PP, and EMP are still insufficiently known, as are the therapies that would be effective in EMD. The aim of this study was to evaluate immunohistochemically the angiogenesis, determined as microvessel density (MVD) and osteopontin expression in PP, of two patients with MM of plasmablastic morphology and an aggressive course of disease. We found high levels of MVD and osteopontin expression in both cases of PP. The role of angiogenesis and osteopontin in EMD should be clarified in future investigations, especially since there are no satisfactory therapeutic protocols for this form of multiple myeloma, and both of these biological factors can be the potential targets of new therapies.

The Protective Benefit of Heme Oxygenase-1 Gene-Modified Human Placenta-Derived Mesenchymal Stem Cells in a N-Nitro-L-Arginine Methyl Ester-Induced Preeclampsia-Like Rat Model: Possible Implications for Placental Angiogenesis.

We previously reported that cytoprotective Heme oxygenase-1, HO-1 (HMOX1) gene-modified human placenta-derived mesenchymal stem cell (HO-1-PMSC) improved placental vascularization in vitro. In the current study, we explored the protective benefit of HO-1-PMSC transplantation in a preeclampsia (PE)-like rat model. A model of PE was successfully constructed by intraperitoneal injection of N-nitro-L-arginine methyl ester (L-NAME). Blood pressure and urinary protein levels were measured. Doppler ultrasound was examined to understand uteroplacental perfusion. ELISA was used to examine the serum levels of VEGF, PlGF, sFlt-1, and sEng. The placentas and fetuses were weighed to verify the improvement in pregnancy outcome. Immunohistochemical and H&E staining was used to detect microvessel density (MVD) in placental tissues and kidney pathology, respectively. The distribution of GFP-labeled PMSC in the placenta were observed under fluorescence microscopy. Blood pressure and proteinuria were reduced and kidney damage was improved. PE rat models treated with PMSC and HO-1-PMSC exhibited an increase in the quality of fetuses and placentas, MVD, VEGF, and PlGF expression, but substantially decreased expression of sFlt-1 and sEng. Doppler ultrasound showed that the placental perfusion was improved. Green fluorescent tracing experiments verified that the cells were successfully transplanted into the placenta and distributed in the blood vessels, indicating that the cells might participate in the process of angiogenesis. These results indicate that therapy with HO-1-PMSC could improve placental vascular dysplasia, increase placental perfusion, control PE symptoms, and promote pregnancy outcome by regulating the balance of angiogenic and antiangiogenic factors or directly participating in the repair of placental vessels in a PE-like rat model.

Down-Regulation of Toll-Like Receptor 5 (TLR5) Increased VEGFR Expression in Triple Negative Breast Cancer (TNBC) Based on Radionuclide Imaging.

In this study, GFP-tagged TNBC 4T1 cells with down-regulated TLR5 expression (TLR5− 4T1) and normal TLR5 expression (TLR5+ 4T1) were constructed, respectively. RT-PCR and Western blot studies showed that down-regulation of TLR5 obviously increased the expression of VEGFR in 4T1 cells. Highly stable radio-probes 125I-anti-TLR5 mAb/125I-VEGF/125I-IgG were obtained with labeling rates over 85% and radiochemical purities above 90%. Among these three probes, 125I−anti−TLR5 mAb and 125I-VEGF were used for specifically imaging TNBC, while 125I-IgG was used for comparison. Whole-body phosphorus autoradiography showed clear imaging at 48 h after injection of 125I-anti-TLR5 mAb and 125I-VEGF also provided clear imaging at 24 h. Biodistribution study demonstrated a higher tumor uptake of 125I-anti-TLR5 mAb in TLR5+ group compared with that in TLR5− group (P < 0.05), whereas tumor uptake of 125I-VEGF in TLR5+ group was lower than that in the TLR5− group (P < 0.05). Immunohistochemical staining suggested that the expression of TLR5 was lower, whereas the expression of VEGFR, CD31, and MVD (microvessel density) was higher in TLR5− tumor-bearing mice. In summary, the down-regulation of TLR5 in TNBC promoted the VEGFR expression and angiogenesis, resulting in the proliferation of TNBC cells. TLR5/VEGF might be a better indicator for monitoring the development of TNBC.

Epithelial-to-Mesenchymal Transition and Neoangiogenesis in Laryngeal Squamous Cell Carcinoma.

Simple summaryThe mechanism of epithelial–mesenchymal transition is fundamental for carcinogenesis, tumor progression, cancer cell invasion, metastasis, recurrence, and therapy resistance, resulting in cellular junction degradation and increased cellular motility. The same factors that drive epithelial cells toward a mesenchymal phenotype may also drive endothelial cells toward a proangiogenic phenotype. This study aimed to investigate a potential interplay between epithelial–mesenchymal transition and angiogenesis in laryngeal carcinoma. In our study, univariate Cox regression identified pN+ status and Slug expression as predictive of disease-free survival, while a trend toward significance emerged for CD105-assessed microvessel density and N-cadherin expression. In the multivariate Cox regression model, pN-status, Slug, and N-cadherin expressions retained their significant values in predicting disease-free survival. Data from our study support the hypothesis of a mutual concurrence of epithelial–mesenchymal transition and angiogenesis in the development of an aggressive phenotype in laryngeal squamous cell carcinoma.The mechanism of epithelial–mesenchymal transition (EMT) is fundamental for carcinogenesis, tumor progression, cancer cell invasion, metastasis, recurrence, and therapy resistance, comprising important events, such as cellular junction degradation, downregulation of epithelial phenotype markers, overexpression of mesenchymal markers, and increase in cellular motility. The same factors that drive epithelial cells toward a mesenchymal phenotype may also drive endothelial cells toward a proangiogenic phenotype. The aim of this exploratory study was to investigate a potential interplay between EMT and angiogenesis (quantified through CD105 expression) in laryngeal carcinoma (LSCC). CD105-assessed microvessel density (MVD) and EMT markers (E-cadherin, N-cadherin, Snail, Slug, Zeb1, and Zeb2) were assessed on 37 consecutive LSCC cases. The univariate Cox regression model identified pN+ status (p = 0.0343) and Slug expression (p = 0.0268) as predictive of disease-free survival (DFS). A trend toward significance emerged for CD105-assessed MVD (p = 0.0869) and N-cadherin expression (p = 0.0911). In the multivariate Cox model, pN-status, Slug, and N-cadherin expressions retained their significant values in predicting DFS (p = 0.0346, p = 0.0430, and p = 0.0214, respectively). Our data support the hypothesis of a mutual concurrence of EMT and angiogenesis in driving LSCC cells toward an aggressive phenotype. To better characterize the predictive performance of prognostic models based on EMT and angiogenesis, further large-scale prospective studies are required.

This Is a Title in Title Case: SDF1,CXCR4 and MVD Play Important Roles in Multiple Myeloma

The myeloid microenvironment of patients with multiple myeloma plays an important role in regulating the occurrence, development, invasion, secretion and angiogenesis of myeloma cells. Chemokines SDF-1 and its receptor CXCR4 form SDF1-CXCR4 biologicalaxis in bone marrow microenvironment. The role and mechanism of SDF1-CXCR4 biologicalaxis in multiple myeloma remain to be studied.We collected 144 cases of hematology bone marrow from January 2013 to June 2017 in Southwest Medical University Affiliated Hospital and the Third People's Hospital of Chengdu, of which 104 cases were multiple myeloma patients, and 40 cases were normal as control. The relationship between SDF1-CXCR4 expression and its clinical features were analyzed. Through these analysis we tried to learn the role of SDF1-CXCR4 in the development, progression and prognosis of multiple myeloma, and hoped to provide evidence for the diagnosis and treatment of multiple myeloma.As a result, the positive expression of SDF1 and CXCR4 in bone marrow biopsy of patients with multiple myeloma was significantly higher than that in the normal control group. More importantly, there was a positive correlation between the expression of SDF1 and CXCR4, indicating that SDF1-CXCR4 in bone marrow microenvironment plays an important role in the occurrence of multiple myeloma.The density of microvessel density (MVD) in the myeloma was also significantly higher than that in the normal control group. The expression of SDF1-CXCR4 in the myeloma group was positively correlated with the expression of MVD. This study demonstrates that multiple myeloma cell growth and microvascular proliferation are significantly correlated in microenvironment. It is suggested that the secretion of chemokine SDF1-CXCR4 in the bone marrow microenvironment of multiple myeloma may promote microvascular proliferation and the growth of tumor cells, which may be the pathogenesis.Multiple myeloma clinical features were analyzed according to SDF1-CXCR4 and MVD expression levels. We found that SDF1-CXCR4 and MVD expression levels were not correlated with gender, age and immunoglobulin typing, but were significantly correlated with multiple myeloma staging in bone marrow tissue of MM patients. But we found that in the later stage of SDF1, the expression level of SDF1、CXCR4 and MVD is higher. This finding further indicates that chemokine SDF1-CXCR4 secretion and microvascular proliferation play important roles in progression of multiple myeloma.The Kplan-Meier analysis on overall survival (OS) and progression-free survival (PFS) was performed according to the levels of SDF1-CXCR4 and MVD expression in multiple myeloma. We discovered that the OS rate and PFS rate were lower in the patients with higher expression of SDF1, CXCR4 and MVD , which indicates that chemokine SDF1-CXCR4 secretion and microvascular proliferation also play important roles in prognosis of multiple myeloma.Our findings reveal that chemokine SDF1-CXCR4 secretion and microvascular hyperplasia in bone marrow microenvironment are essential in the occurrence, development and prognosis of multiple myeloma. The higher secretion of chemokine SDF1-CXCR4 suggests the worse progression and poor prognosis. Chemokine SDF1-CXCR4 is expected to become a new target for MM treatment. DisclosuresNo relevant conflicts of interest to declare.

Impact of Cox2, CD163, and Microvessel Density Expression on the Prediction of Relapse and Patients’ Outcome in Classical Hodgkin Lymphoma

Background: Tumor microenvironment, specifically tumor-associated macrophages, plays an important role in tumor initiation and progression. CD163 has been recognized as a valuable specific macrophage marker. Cyclooxygenase-2 (Cox2) plays a role in tumor progression. CD31 is reliable for estimation of the density of microvesseles (MVD), which has prognostic importance in several malignant tumors. Thus, the current study was conducted to test the association between CD163, Cox2, and CD31 expression with the prognosis of classical Hodgkin lymphoma (cHL) patients and their potential correlation with clinicopathological variables. Method: CD163, Cox2, and CD31 expressions were examined in newly diagnosed patients with cHL through immunohistochemistry on tissue biopsy and the results were correlated with the patients’ outcome after the median follow-up, which was about 35 months. Results: 104 patients were included in this study. High CD163 was found in 32.7 % of the patients. Cox2 was positive in 42.3% of them. CD 31 with high MVD (≥10%) was found in 51% of the subjects. A significant association was detected between CD163 and Cox2 with tumor stage (P=0.001, and P= 0.001) and IPS score. Regarding CD31, we could not find any significant associations with disease parameters, except with histological subtype (P=0.001). A significant relationship was observed between Cox2 and CD163 expression and the relapse rate (P=0.001, P=0.01 respectively). Regarding survival, only Cox2 showed a significant association with disease-free survival (DFS) (P=0.0379). Conclusion: These findings suggested that Cox2 and CD163 expression can be used as predicator for early relapse and as new therapeutic targets in cHL.

The role of c-Met and VEGFR2 in glioblastoma resistance to bevacizumab

Dismal prognosis of glioblastoma (GBM) prompts for the identification of response predictors and therapeutic resistance mechanisms of current therapies. The authors investigated the impact of c-Met, HGF, VEGFR2 expression and microvessel density (MVD) in GBM patients submitted to second-line chemotherapy with bevacizumab. Immunohistochemical expression of c-Met, HGF, VEGFR2, and MVD was assessed in tumor specimens of GBM patients treated with bevacizumab, after progression under temozolomide. Survival analysis was evaluated according to the expression of the aforementioned biomarkers. c-Met overexpression was associated with a time-to-progression (TTP) after bevacizumab of 3 months (95% CI, 1.5–4.5) compared with a TTP of 7 months (95% CI, 4.6–9.4) in patients with low or no expression of c-Met (p = 0.05). VEGFR2 expression was associated with a TTP after bevacizumab of 3 months (95% CI, 1.8–4.2) compared with a TTP of 7 months (95% CI, 5.7–8.3) in patients with no tumoral expression of VEGFR2 (p = 0.009). Concomitant c-Met/VEGFR2 overexpression was associated with worse overall survival (13 months) compared with concomitant c-Met/VEGFR2 negative expression (19 months; p = 0.025). Our data support the hypothesis that c-Met and VEGFR2 overexpression have a role in the development of glioblastoma early resistance and might predict poorer responses to anti-angiogenic therapies.

Acidosis induces synovial fibroblasts to release vascular endothelial growth factor via acid-sensitive ion channel 1a

AbstractAcid-sensitive ion channel 1a (ASIC1a) is a member of the extracellular H+ activated cation channel family. Studies have shown that tissue acidification contributes to the formation of microvessels in rheumatoid arthritis (RA) synovial tissue, but its underlying mechanisms remain unclear. The purpose of this study was to investigate the role of tissue acidification in microvascular formation of arthritic synovial tissue and the effect of ASIC1a on vascular endothelial growth factor (VEGF) release from arthritic synovial tissue. Our results indicate that ASIC1a expression, VEGF expression, and microvessel density (MVD) are elevated in RA synovial tissue and adjuvant arthritis (AA) rat synovial tissue. When AA rats were treated with ASIC1a-specific blocker psalmotoxin-1 (PcTx-1), the expression of ASIC1a, VEGF expression, and MVD were all reduced. Acidification of RA synovial fibroblasts (RASF) can promote the release of VEGF. PcTx-1 and ASIC1a-short hairpin RNA can inhibit acid-induced release of VEGF. In addition, the ASIC1a overexpression vector can promote acid-induced VEGF release. This indicates that extracellular acidification induces the release of VEGF by RASF via ASIC1a. These findings suggest that blocking ASIC1a mediates the release of VEGF from synoviocytes may provide a potential therapeutic strategy for RA therapy.Both ASIC1a and VEGF are highly expressed in rheumatoid arthritis synovial tissue and are associated with vascular disease. Interfering with ASIC1a in vitro using silencing, blocking, and overexpression interferes with the release of VEGF under acid stimulation. Blocking ASIC1a in the articular cavity of rats with adjuvant arthritis not only reduces the expression of VEGF in the synovium, but also reduces the proliferation and lesions of blood vessels and interferes with the development of the disease.

Microvessels in tumor tissue in patients with central nervous system tumors

Angiogenesis plays an essential role in the development of a tumor. The most preferred histological evaluation method of this process is the determination of microvessel density (MVD) in the biopsy. Numerous studies report the possibility of using various MRI techniques for imaging the network of neoplastic vessels and determining the angiogenesis in them, as well as the subsequent analysis of the obtained images. In this study, a classic MRI imaging with intravenous contrast agent administration was used, and in order to ensure objectification and repeatability, and to eliminate human error that may occur in classic MRI image analysis, an automatic evaluation algorithm was used to determine the degree of angiogenesis, which was then presented in the form of a statistical pixels analysis. The study involved 48 adult patients of both sexes, treated surgically at the Department of Neurosurgery, Nicolaus Copernicus University Collegium Medicum in Bydgoszcz, due to a solid brain tumor. The highest mean density of MVD microvessels in tumor tissue was observed in high-grade gliomas. The high level of MVD expression in tumors of the central nervous system showed a positive correlation with the high degree of tumor malignancy. Based on the conducted research, it was concluded that the determination of the level of MVD expression in the tumor tissue is an additional prognostic factor in neoplasms of the central nervous system. The comparison of the results obtained with the tests performed routinely translates into a more accurate selection of treatment and assessment of the prognosis for survival.

Sorafenib blocks the activation of the HIF-2α/VEGFA/EphA2 pathway, and inhibits the rapid growth of residual liver cancer following high-intensity focused ultrasound therapy in vivo

BackgroundInsufficient high-intensity focused ultrasound (HIFU) can promote the rapid progression of the residual tumor through the hypoxia inducible factor-2α +(HIF-2α)/vascular endothelial growth factor A (VEGFA)/ephrin type-A receptor 2 (EphA2) pathway. Although sorafenib has been shown to significantly improve the survival of patients with advanced liver cancer, the use of sorafenib in residual tumor tissues following HIFU has rarely been elucidated. Thus, this study aimed to investigate the potential adjuvant therapeutic effects of sorafenib following HIFU in order to reduce the relapse rate following insufficient HIFU. MethodsXenograft tumors were established using nude mice injected with liver cancer cells. At approximately 4 weeks after the inoculation of the tumor cells (tumors reached 1.3–1.5 cm), all mice were randomly divided into 3 groups as follows: i) The control group (no treatment); ii) the HIFU-alone group, and iii) the combination group (HIFU + sorafenib), with 6 mice per group. The residual tumor volume was determined among the different treatment groups. The protein expression levels of HIF-2α, VEGFA and EphA2 were determined by immunohistochemistry and western blotting, and the mRNA levels were detected by RT-qPCR. The microvessel density (MVD) was calculated by CD31 immunohistochemistry staining. ResultsThe results revealed that by comparing the control group, insufficient HIFU promoted HIF-2α, VEGFA and EphA2 expression (P < 0.05). Compared with the HIFU-alone group, the protein and mRNA levels of HIF-2α, VEGFA and EphA2 were markedly decreased in the group that received combined treatment with HIFU and sorafenib (P < 0.05). Similar results were obtained for MVD expression. Synergistic tumor growth inhibitory effects were also observed between the control group and HIFU group (P < 0.05). ConclusionsThe findings of this study demonstrate that the expression of HIF-2α, VEGFA and EphA2 can be inhibited by sorafenib, and that sorafenib is likely to provide an effective adjunct treatment for patients with HCC following HIFU ablation.