Role of heat shock protein 27 in transforming growth factor-β-stimulated vascular endothelial growth factor release in osteoblasts

We have previously reported that transforming growth factor-β (TGF-β) stimulates heat shock protein?27 (HSP27) induction via p44/p42 mitogen-activated protein (MAP) kinase, p38 MAP kinase and stress-activated protein kinase/c-Jun N-terminal kinase in osteoblast-like MC3T3-E1 cells, and that the release of vascular endothelial growth factor (VEGF) is induced by TGF-β in these cells. In the present study, we investigated the effect of HSP27 knockdown on the TGF-β-stimulated VEGF release in these cells. Gene silencing using short interfering RNA against HSP27 (HSP27-siRNA) significantly suppressed the TGF-β-induced VEGF release. Immunofluorescence microscopy also revealed that HSP27-siRNA suppressed the TGF-β-stimulated VEGF induction as well as the reduction of HSP27 induction in these cells. However, the mRNA expression of VEGF stimulated by TGF-β was not reduced even in cells transfected with HSP27-siRNA. These results strongly suggest that HSP27 induction is critical for TGF-β-induced VEGF release in osteoblasts.     

Dual functionalization of titanium with vascular endothelial growth factor and β-defensin analog for potential application in keratoprosthesis

Functionalization of material surfaces can improve their biointegration and bactericidal effect. To expand the biomedical applications of titanium in artificial cornea implantation surgery, titanium alloy substrates were coated with polydopamine and dual bound with recombinant vascular endothelial growth factor (VEGF) and anti-microbial peptide (AMP), SESB2V. Successful chemical binding was assessed with attenuated total reflectance-Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Coating thickness was assessed by atomic force microscopy. Cellular studies revealed that the functionalized substrates displayed the abilities to enhance primary human corneal fibroblast adhesion, proliferation, and viability. Angiogenesis assay with human mesenchymal stem cells was used to verify the biological functions of immobilized VEGF while bactericidal assay was evaluated for the anti-microbial activities of immobilized SESB2V peptide. We found that the titanium surface that was sequentially functionalized with VEGF and SESB2V had enhanced fibroblast proliferation and anti-microbial properties. The incorporation of such peptides into an artificial cornea implant is important for implant-tissue integration and wound healing. This may improve implant integration and reduce the risk of device infection following artificial cornea implantation.     

Is insulin-like growth factor binding protein 2 associated with metastasis in lung cancer?

Insulin-like growth factor binding protein 2 (IGFBP2) is involved in the progression of many epithelial cancers. However, its role in non-small cell lung cancer (NSCLC), another type of epithelial cancer, remains unclear. We detected IGFBP2 expression using immunohistochemistry in surgically resected tumors from 110 NSCLC patients, 37 of which had metastases. The positive rate of IGFBP2 expression was compared between the metastatic and the non-metastatic group, and correlations of IGFBP2 expression with metastasis and overall survival were analyzed. We also investigated the expression of IGFBP2 in microvesicles (MVs) collected from primary lung cancer cell cultures, and in different locations of newly resected NSCLC tumors, using immunoblotting. The overall positive rate of IGFBP2 expression in lung cancer was 51.8 % and it was significantly higher in the metastatic group than in the non-metastatic group (70.3 and 42.5 % respectively, p < 0.01). And the higher the lymph node stage, the higher the positive rate. Cytoplasmic expression was predominant in the majority of the tumors. Based on multivariate regression analysis, IGFBP2 was correlated with metastasis and poor overall survival (Hazard ratio: 3.56 and 3.23 respectively). IGFBP2 was detectable in the MVs collected from IGFBP2 positive cell lines, and its expression was most abundant in the marginal region of the newly resected tumors. IGFBP2 is associated with metastasis and poor survival of lung cancer. Its presence in MVs and high abundance in the marginal region of tumors suggest that its association with metastasis may be related to tumor microenviroment remodeling in NSCLC.     

Heat shock protein 90-β over-expression is associated with poor survival in stage I lung adenocarcinoma patients

Heat shock protein 90-beta (Hsp90-β) is associated with cell proliferation, differentiation and apoptosis and has been investigated as a prognostic factor in many cancers. However, Hsp90-β protein expression in lung adenocarcinoma (ADC) has not been thoroughly elucidated. The aim of this study was to determine the relationship between Hsp90-β expression, clinicopathological parameters and prognosis in lung adenocarcinomas. Seventy-five surgically resected lung adenocarcinomas and matched normal lung tissue samples were obtained to construct a tissue microarray (TMA), including 44 stage IA-IB cases. Then, Hsp90-β protein expression level in lung tissue was evaluated by immunohistochemistry. Kaplan-Meier survival analysis with a Log-rank significance test was used to estimate the survival differences among subgroups according to Hsp90-β expression in lung ADC tissues using SigmaPlot/SigmaStat v10 and 3.5, respectively. Hsp90-β protein expression was significantly upregulated in lung ADC tissues compared to that in the matched normal alveoli (P<0.001) and was associated with tumor differentiation (P<0.001). Furthermore, Hsp90-β over-expression was correlated with poor survival in stage I patients (P=0.026). Increased Hsp90-β expression was associated with reduced overall survival (HR, 2.440; 95% confidence interval, 1.076-5.530; P=0.033). To conclude, our data demonstrated that Hsp90-β protein was over-expressed in lung ADC tumor tissues and was associated with poor outcomes in early stage ADC patients and low pathological grade tumors. These data suggest that Hsp90-β could be a clinically useful biomarker for the prognosis of ADC and an effective anticancer target.     

Hypoxia,hypoxia-inducible factor-1α and vascular endothelial growth factor in a murine model of Schistosoma mansoni infection

Schistosomiasis mansoni is a chronic parasitic disease where much of the symptomatology is attributed to granuloma formation, an immunopathological reaction against Schistosoma eggs. To more clearly understand the immunopathology of schistosomiasis, the tissue microenvironment generated by S. mansoni infected mice was investigated. Using the hypoxia marker pimonidazole, we provide immunohistochemical evidence that hypoxia occurred in inflammatory cells infiltrated around the eggs and cells surrounding granulomas in the liver, intestine, spleen and lungs of infected mice. Hypoxia-inducible factor-1α (HIF-1α) was mainly expressed in inflammatory cells surrounding the eggs and in hepatocytes surrounding cellular and fibrocellular granulomas in infected mouse liver. HIF-1α expression was also verified in granulomas in the other tissues tested (intestine, spleen and lungs). Vascular endothelial growth factor (VEGF) expression was observed in the extracellular space surrounding inflammatory cells in liver granuloma. The VEGF expression pattern verified in infected mouse liver was very similar to that observed in the other tissues tested. A strong positive correlation occurred between pimonidazole binding and HIF-1α and VEGF expression in the tissues tested, except for lung. This work is the first evidence that infection by a helminth parasite, S. mansoni, produces a hypoxic tissue microenvironment and induces HIF-1α and VEGF expression.     

β-arrestin 2 is associated with multidrug resistance in breast cancer cells through regulating MDR1 gene expression

Mutidrug resistance (MDR) severly blocks the successful management of breast cancer. Overexpression of MDR1/p-gp accounts for the major factor in the development of MDR. β-arrestin 2 has been reported to widely involve in multiple aspects of tumor development. In order to verify whether β-arrestin 2 regulates mutidrug resistance in breast cancer, we analyzed the protein expression levels of β-arrestin 2 and MDR1/p-gp by immunohistochemistry in 106 paraffin-embedded human breast tissue samples. There was a positive correlation between β-arrestin 2 and MDR1/p-gp protein expression (P = 0.016). Changes in MDR1/p-gp mRNA and protein levels were examined by quantitative real-time reverse polymerase chain reaction (qRT-PCR) and western blotting. Silencing of β-arrestin 2 evidently down-regulated the expression of MDR1/p-gp in transfected ADM cells. In contrast, overexpression of β-arrestin 2 had the opposite changes in MDA-MB-231 and MCF-7 cells. MTS assay revealed that silencing of β-arrestin 2 increased the sensitivity to anti-cancer drugs to some extent. On the other hand, overexpression of β-arrestin 2 had the opposite effects. Our above data demonstrate that β-arrestin 2 plays a vital role in the regulation of MDR1/p-gp expression in Breast cancer.     

The rs3761548 FOXP3 variant is associated with multiple sclerosis and transforming growth factor β1 levels in female patients

Inflammation Research - The aim of this study was to evaluate the association between rs3761548 FOXP3 (-3279 C?&gt;?A) variant and multiple sclerosis (MS), disability, disability...     

Tanshinone IIA-induced attenuation of lung injury in endotoxemic mice is associated with reduction of hypoxia-inducible factor 1α expression

Inhibiting hypoxia-inducible factor (HIF)-1α activity has been proposed as a novel therapeutic target in LPS-induced sepsis syndrome. We have reported that tanshinone IIA (TIIA) can reduce LPS-induced lethality and lung injury in mice, but the precise mechanisms have not been fully described. Therefore, the present study investigated whether the protective effect of TIIA was related to the inhibition of LPS-induced HIF-1α expression and what mechanisms accounted for it. This study showed that TIIA pretreatment improved LPS-induced biochemical and cellular changes and reduced the production of inflammatory cytokines. Pretreatment with TIIA decreased LPS-induced HIF-1α expression in vivo and in vitro. TIIA did not affect the LPS-induced HIF-1α mRNA level but inhibited HIF-1α protein translation by the inhibition of the PI3K/AKT and MAPK pathways and related protein translational regulators, such as p70S6K1, S6 ribosomal protein, 4E-BP1, and eIF4E, and promoted HIF-1α protein degradation via the proteasomal pathway in LPS-stimulated macrophages. These observations partially explain the antiinflammatory effects of TIIA, which provides scientific basis for its application for the treatment of acute lung injury/acute respiratory distress syndrome or sepsis.     

Expression of vascular endothelial growth factor (VEGF), hypoxia inducible factor 1 alpha (HIF-1α), and transforming growth factors β1 (TGFβ1) and β3 (TGFβ3) in gestational trophoblastic disease

The aim of this study was to investigate the relationship between the expression of vascular endothelial growth factor (VEGF), transforming growth factor β (TGF-β1 and TGF-β3), and hypoxia inducible factor 1 alpha (HIF-1α) in gestational trophoblastic diseases to highlight the possible histogenesis.     

Osteopontin is associated with decreased apoptosis and αv integrin expression in lung adenocarcinoma

Osteopontin (OPN) is a glycoprotein involved in invasion, progression and metastasis of many carcinomas. It contains several functional domains including binding sites for αv integrins, cell surface molecules playing a major role in mediating cell migration and adhesion. The aim of the study was to evaluate the expression of osteopontin in human non-small cell lung cancer (NSCLC) and to determine its possible prognostic significance as well as relation to apoptosis and αv integrin expression. We analyzed 111 surgically resected NSCLC for immunohistochemical expression of OPN and αv integrin. OPN expression was compared to apoptotic rate and clinicopathological parameters such as tumor size, histological grade, lymph node status, pT, and TNM stage. Apoptotic rate was measured by TUNEL staining method. OPN expression in NSCLC was significantly higher in lung adenocarcinomas (AC) then in squamous cell carcinomas (p < 0.001). There was no correlation between OPN expression and clinicopathological parameters. The level of OPN expression in AC was associated with decreased apoptotic activity of tumor cells (p = 0.006), and correlated with αv integrin expression (p = 0.048), particularly in low stage tumors (p = 0.013). Prolonged tumor cell survival in lung AC due to OPN and αv integrin overexpression may have an impact on tumor progression and resistance to therapy.     

Jab1 expression is associated with TGF-β1 signaling in chronic rhinosinusitis and nasal polyposis

Jab1, which is a fifth component of COP9 signalosome, plays an essential role in cell growth and proliferation. Jab1 is also shown to regulate transforming growth factor-beta (TGF-β) signaling in carcinoma cells. The aim of the present study was to investigate the expression and the correlation of Jab1 and TGF-β1 in chronic rhinosinusitis and nasal polyposis. Here, we show the elevated expression of Jab1 and TGF-β1 in diseased mucosa without nasal polyps and a correlation between Jab1 and TGF-β1 expression. Forty-six samples (26 patients with nasal polyps, 10 patients with chronic rhinosinusitis and 10 control subjects) were included to this study. Immunohistochemistry and Western blotting were performed for the assessment of Jab1 and TGF-β1 localization and the expression of proteins. Double staining of both proteins showed that Jab1 and TGF-β1 were colocalized in the epithelium, inflammatory cells and the vascular endothelium of nasal mucosa. There was a significant increase in the expression of TGF-β1 and Jab1 in patients without nasal polyps and a significant decrease in patients with nasal polyps compared to controls. Moreover, correlation was detected between the expression of Jab1 and TGF-β1 in chronic rhinosinusitis and nasal polyposis. Our results demonstrate that chronic rhinosinusitis is characterized by elevated expression of Jab1 and TGF-β1 compared to nasal polyposis and Jab1 may play a vital role in the pathogenesis of both chronic rhinosinusitis and nasal polyposis.     

Levels and patterns of expression of hypoxia-inducible factor-1α, vascular endothelial growth factor, glucose transporter-1 and CD105 in adenoid cystic carcinomas with high-grade transformation

Costa A F, Tasso M G, Mariano F V, Soares A B, Chone C T, Crespo A N, Fresno M F, Llorente J L, Suárez C, de Araújo V C, Hermsen M & Altemani A
(2012) Histopathology  60, 816–837 Levels and patterns of expression of hypoxia‐inducible factor‐1α, vascular endothelial growth factor, glucose transporter‐1 and CD105 in adenoid cystic carcinomas with high‐grade transformation Aims: To compare the expression of proteins regulated by hypoxia between adenoid cystic carcinoma (ACC) with and without high‐grade transformation (HGT). Methods and results: In eight ACC–HGT and 18 ACC without HGT, expression of hypoxia‐inducible factor‐1 (HIF‐1α), vascular endothelial growth factor (VEGF), glucose transporter‐1 (GLUT‐1) and microvascular density (MVD) by CD105 (a hypoxia‐inducible protein expressed in angiogenic endothelial cells) was determined. Expression levels of HIF‐1α and VEGF as well as CD105‐MVD did not differ significantly between: (i) transformed and conventional areas (TA and CA, respectively) of ACC–HGT, (ii) CA and ordinary ACC. HIF‐1α was detected in 100% of cases and presented a diffuse expression pattern. No significant association was found between levels of HIF‐1α expression and tumour size, metastasis and recurrence. GLUT‐1 showed a prostromal expression pattern and was observed exclusively in TA (three of six cases) and in only three of 14 ACC. Conclusions: Both the absence of significant alterations in levels of expression of HIF‐1α, VEGF and CD105 and the patterns of expression of HIF‐1α and GLUT‐1 suggest that hypoxia may not play a key role in the process of high‐grade transformation of ACC. Although HIF‐1α expression is a common finding in ACC, it cannot be used as a marker of tumour aggressiveness.     

Loss of nuclear expression of Krüppel-like factor 4 is associated with poor prognosis in patients with oral cancer

Krüppel-like factor 4 is not only involved in cell proliferation but also affects cell differentiation and extracellular matrix production via positive and negative regulation of the expression of a wide range of genes. To our knowledge, little information is available regarding the role of Krüppel-like factor 4 in oral squamous cell carcinoma. In this study, we investigated the associations between Krüppel-like factor 4 expression and clinical parameters of oral cancer using immunohistochemical assays in 215 surgical specimens. Compared with positive nuclear Krüppel-like factor 4 expression, we observed that negative nuclear Krüppel-like factor 4 expression was significantly associated with an advanced cancer stage (P = .046), a high tumor recurrence rate (P = .009), and a worse 3-year survival rate in patients with oral cancer (P = .046). Nuclear expression of Krüppel-like factor 4 was shown to have an inverse relationship with Ki67 expression (P = .046). Patients with negative nuclear expression of Krüppel-like factor 4 had significantly worse overall survival rates as defined by the log-rank test (P = .014). Patients with oral cancer with negative nuclear Krüppel-like factor 4 expression in tumor cells had poor prognoses and a 2.5-fold higher death risk. Compared with disease stage (P = .025), negative nuclear Krüppel-like factor 4 expression (P = .006) was an independent prognostic factor. Our results revealed that the loss of nuclear expression of Krüppel-like factor 4 is significantly associated with aggressive clinical manifestations and might be an adverse survival factor.     

Are epidermal growth factor and transforming growth factor responsible for pseudoepitheliomatous hyperplasia associated with granular cell tumors?

Granular cell tumors (GCT) are uncommon benign neoplasms that have a predilection for the head and neck region. These tumors can frequently be associated with pseudoepitheliomatous hyperplasia (PEH), which in turn may be mistaken for squamous cell carcinoma. Although epidermal growth factors are overexpressed in squamous cell carcinomas of the head and neck, their presence in PEH, especially its relation to GCT, is unknown. We hypothesize that the expression of epidermal growth factor receptor (EGFR), epidermal growth factor (EGF), and transforming growth factor alpha (TGFalpha) in GCT have a role in the development of PEH overlying some GCT. Sections from 13 cases of GCT (five with overlying PEH) were examined histologically and evaluated immunohistochemically using monoclonal antibodies for EGFR, EGF, and TGFalpha. These were compared with nine cases of PEH independent of GCT. Two of five GCT with overlying PEH and two of six GCT without overlying PEH stained positively for TGFalpha. None of the GCT stained with EGFR or EGF. All cases of PEH, whether or not associated with GCT, were reactive for EGFR and EGF. Four of the five cases of PEH overlying GCT stained with TGFalpha. The staining pattern and intensity of all three antibodies were comparable to that of the adjacent normal squamous mucosa. Among the three antibodies, only TGFalpha in GCT appears to be related to the development of PEH. Epidermal growth factor receptor and EGF do not seem to be directly involved. The reason of PEH formation associated with GCT in the absence of growth factors is unknown.     

Angiogenesis and expression of vascular endothelial growth factor, tumour necrosis factor-α and hypoxia inducible factor-1α in canine renal cell carcinoma

The aim of the present study was to determine the distribution and characteristics of microvessels in various histological types of canine renal cell carcinoma (RCC). The study compared microvessel density (MVD) and distribution of blood vessels according to histological type and evaluated the presence of angiogenesis-related proteins. Nine archival samples of canine RCC were studied. MVD was calculated as the mean number of blood vessels per mm(2). The diameter of blood vessels was calculated by determining either the length of the long axis of blood vessels (diameter(max)) or the mean distance from the centre of each blood vessel to the tunica adventia (diameter(mean)). A significant difference in MVD was evident between RCCs and normal kidneys (46.6 ± 28.0 versus 8.4 ± 2.2 microvessels/mm(2)). Diameter(max) in canine RCCs (34.1 ± 14.7 μm) was also significantly different from normal canine kidney (23.2 ± 3.4 μm). Vascular endothelial growth factor (VEGF) was expressed by tumour cells and vascular endothelial cells and tumour necrosis factor (TNF)-α expression was observed in vascular endothelial cells in both neoplastic and normal kidney. Although VEGF is involved in angiogenesis and correlates with tumour stage of development, no correlation was found between VEGF expression and MVD. Tumour-associated macrophages expressing TNF-α and hypoxia inducible factor 1α were identified in peritumoural tissue and may play an important role in angiogenesis.