[Show abstract][Hide abstract] ABSTRACT: Gelsolin (GSN) is one of the most abundant actin-binding proteins, and is involved in several pathological processes, including Alzheimer's disease, cardiac injury and cancer. The aim of the present study was to assess the effect of GSN on the growth and motility of oral squamous cell carcinoma Tca8113 cells. The overexpression vector pcDNA3.1-GSN was transfected into Tca8113 cells and the stable GSN overexpression cell line was identified based on G418 antibiotic selection. The effect of GSN overexpression on the proliferation, apoptosis, migration and invasion of Tca8113 cells was examined using a cell counting kit-8 assay, flow cytometry and Transwell assays. The results revealed that GSN overexpression significantly promoted the cell proliferation and apoptosis of Tca8113 cells. In addition, Transwell assays demonstrated that the migration and invasion abilities of Tca8113 cells were enhanced by GSN overexpression. Therefore, the upregulation of GSN promotes cell growth and motility, indicating that it may perform a vital function in the progression of human oral cancers.
[Show abstract][Hide abstract] ABSTRACT: To mimic oral squamous cell carcinoma (OSCC) cell hypoxia by using chemical agent CoCl2 and to investigate its biological behaviour.
Oral squamous cell carcinoma cell lines HSC-3 and SCC-4 were exposed to different concentration of CoCl2. HSC-3 and SCC-4 cells were treated with 50, 100, 150, 200 µmol/L CoCl2. Expression of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and B-cell lymphoma-2 (BCL-2) were measured by real time polymerase chain reaction (PCR) and Western blotting in both mRNA and protein level. Cell proliferation, cell apoptosis and cell cycle were detected to analyze its biological behaviour. Both wound healing and Transwell assay were applied to test the ability of cell igration.
The result showed that after treatment of 150 µmol/L CoCl2 for 24 h, mRNA level of HIF-1α, VEGF and Bcl-2 was increased by 6.00 ± 0.20, 5.40 ± 0.40, 5.40 ± 0.30 (SCC-4); 5.60 ± 0.30, 5.20 ± 0.60, 5.80 ± 0.40(HSC-3). OSCC cells treated with 150 µmol/L CoCl2 for 24 h were collected. Compared with control group, the growth rate of cells was significantly decreased, P value was less than 0.05 (when HSC-3, SCC-4 cultured for 2 and 3 days). The apoptosis of OSCC cells was increased when treated with 150 µmol/L CoCl2 for 24 h:HSC-3 2.25% (control group) and 5.82% (treatment group); SCC-4 2.58% (control group) and 10.27% (treatment group). The migration ablility of OSCC cells was decreased when using 150 µmol/L CoCl2 for 24 h. The migration area ratio was (31.5 ± 2.3) % (HSC-3), (29.1 ± 1.5) % (SCC-4) in control group and (18.3 ± 1.9) % (HSC-3), (13.2 ± 0.8)% (SCC-4) in treatment group (P < 0.05).
The hypoxic cell model of OSCC could be induced by CoCl2. The expression level of hypoxic markers was up regulated significantly and the cells biological behaviour changed including decreased cell proliferation, increased apoptosis and decreased migration.
No preview · Article · Mar 2015 · Zhonghua kou qiang yi xue za zhi = Zhonghua kouqiang yixue zazhi = Chinese journal of stomatology
[Show abstract][Hide abstract] ABSTRACT: Objective
The aim of this study is to investigate the expression and function of TLR3 in oral squamous cell carcinoma (OSCC).
We first assessed TLR3 expression in 20 cases of primary OSCC tissue. Then two OSCC cell lines SCC4 and CAL27 were used for further study. Poly(I:C) was used to activate TLR3 expressed by OSCC. Then changes of cytokines expression, cell viability, apoptosis and migration in OSCC were investigated.
TLR3 was present in both OSCC tissue and two OSCC cell lines. Poly(I:C) stimulated robust responses in OSCC: up-regulated cytokines expression; decreased cell viability by suppressing cell proliferation and inducing apoptosis; and decreased cell migration. Poly(I:C)-TLR3-induced OSCC cell apoptosis was caspase-3-dependent.
The present study indicated that TLR3 might impact OSCC development and should be considered as a target for future OSCC immunotherapy.
No preview · Article · Jul 2014 · Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology
[Show abstract][Hide abstract] ABSTRACT: We sought to investigate the role and diagnostic value of microRNA 155 (miR-155) in OSCC patients.
Using real-time quantitative polymerase chain reaction analysis, miR-155 expression levels were assessed in OSCC cell lines and a cancerous HB cell line. The correlation between miR-155 expression level and clinical parameters was analyzed in 46 patients with OSCC. In addition, the effects of miR-155 on OSCC cell proliferation were evaluated by modulating its expression using an miR-155 mimic and antisense miR-155.
Significant upregulation of miR-155 was found in OSCC cell lines and in tissues of patients with OSCC. The receiver operator characteristic analysis indicated fair-to-good predictability. Overexpression of miR-155 correlated with the histologic grade (P = .033), and the upregulation of miR-155 enhanced OSCC cell proliferation.
In OSSC, upregulation of miR-155 correlated with the histologic grade and can be used as a potential prognostic biomarker.
[Show abstract][Hide abstract] ABSTRACT: The successful treatment of cancer with dendritic cell (DC) tumor vaccine is highly dependent on the efficacy of antigen presentation and T cell activation. In the present study, a novel vaccine of DCs fused with autologous tumor cells was introduced, which had a marked ability to suppress head and neck carcinoma. DCs generated from the bone marrow of mice were fused with an autologous tumor cell line using polyethylene glycol (PEG). To observe the fused cells, confocal microscopy and FACS analysis were performed. Subsequently, the activation and proliferation of T cells, as well as animal experiments, were examined. The efficiency of DC/tumor fusion was 18.03% and T cells were well-activated by the hybrids. The volumes of tumors on the tumor-bearing mice were controlled, survival time of tumor-bearing mice was prolonged and the level of IFN-γ in serum was significantly increased compared with the control group and lysate-pulsed DC group. The results indicate that the DC/tumor fusion vaccine appears to be more effective than DCs pulsed with tumor lysate for the treatment of head and neck carcinoma, which may be useful in future clinical studies.
[Show abstract][Hide abstract] ABSTRACT: The successful biotherapy of carcinoma with dendritic cell (DC) vaccines pivotally relies on DCs' migratory capability into lymph tissues and activation of T cells. Accurate imaging and evaluation of DC migration in vivo have great significance during antitumor treatment with DC vaccine. We herein examined the behavior of DCs influenced by synthetic superparamagnetic iron oxide (SPIO) nanoparticle labeling.
γ-Fe2O3 nanoparticles were prepared and DCs, which were induced from bone marrow monocytes of enhanced green fluorescent protein (EGFP) transgenic mice, were labeled. The endocytosis of the SPIO, surface molecules, cell apoptosis and fluorescence intensity of EGFP-DCs were displayed by Prussian blue staining and flow cytometry (FCM), respectively. After EGFP-DCs, labeled with SPIO, were injected into footpads (n = 5) for 24 hours, the mice were examined in vivo by optical imaging (OPI). Meanwhile, confocal imaging and FCM were applied, respectively, to detect the migration of labeled DCs into draining lymph nodes.
Nearly 100% of cells were labeled by the SPIO, in which the intracellular blue color gradually deepened and the iron contents rose with the increase of labeling iron concentrations. In addition, cell apoptosis and the surface molecules on DCs were at similar levels after SPIO labeling. After confirming that the fluorescence intensity of EGFP on DCs was not influenced by SPIO, the homing ability of EGFP-DCs labeled with SPIO displayed that the fluorescence intensity and the ratios of EGFP-DCs in draining lymph nodes were gradually decreased with the increase of labeling iron concentrations.
The synthetic SPIO nanoparticles possess perfect labeling ability and biocompatibility. Moreover, DCs labeled with a low dose of SPIO showed stronger migratory capability in vivo.
Preview · Article · Oct 2013 · International Journal of Nanomedicine
[Show abstract][Hide abstract] ABSTRACT: Expression of the transcription factor hypoxiainducible factor 1 (HIF-1) plays a key role in cellular adaptation to hypoxia, particularly in relation to tumour angiogenesis. Expression of the HIF-1α subunit is responsive to changes in oxygen levels. Overexpression of HIF-1α has been reported to be associated with a poor prognosis in a variety of malignant tumours. The objective of this study was to investigate whether the expression of HIF-1α in tongue carcinoma was associated with established clinicopathological features. Tumour specimens from 120 patients with histologically-proven, surgically-treated tongue carcinoma were examined by immunohistochemical staining for expression of HIF-1α. The mRNA levels of HIF-1α were measured in 45 fresh, paired samples of tongue carcinoma and corresponding adjacent normal tissues using quantitative RT-PCR (qRT-PCR). HIF-1α was found to be frequently overexpressed in tumours in a hypoxia-independent manner. The expression of HIF-1α correlated with the five-year survival rate (P<0.01) and disease-free period (P<0.01). Increased expression of HIF-1α correlated significantly with clinical stage (P=0.002) and lymph node metastasis (P=0.034). Compared with paired normal tissues, HIF-1α mRNA levels were significantly increased in carcinoma of the tongue. A positive correlation was observed between HIF-1α mRNA levels and pathological differentiation grade. A significant difference in the levels of HIF-1α expression was detected between groups of patients with lymph node metastases and patients with no metastases. These results indicate that overexpression of HIF-1α may be an indicator of poor prognosis in carcinoma of the tongue. The expression of HIF-1α may be associated with lymph node metastasis.
[Show abstract][Hide abstract] ABSTRACT: Vascular endothelial growth factor (VEGF) is a tumor angiogenesis factor that is important in immune regulation. In our previous study, we found that VEGF expression in the peripheral blood and neoplasm nest from patients with oral squamous cell carcinoma (OSCC) was positively correlated with the course of disease, while an inverse correlation between VEGF expression and dendritic cells (DCs) was identified in the peripheral blood. Therefore, in the present study, we investigated whether inhibition of human VEGF in the human tongue carcinoma cell line Tca8113 had effects on the activity of monocyte-derived DCs. We knocked down the expression of human VEGF in Tca8113 cells using the small interfering RNA (siRNA) technique. Tca8113 cells pre-transfected with siRNA targeting VEGF were co-cultured with monocyte‑derived immature and mature DCs. Cell proliferation was evaluated by a WST-8 assay. Cell apoptosis, cell cycle and cell phenotypes were determined by flow cytometry. The data revealed that downregulation of the human VEGF significantly inhibited the proliferation of Tca8113 cells and increased apoptosis. Inhibition of human VEGF arrested the cell cycle of Tca8113 cells at the G0/G1 phase. Our results showed that the co-culture of DCs with Tca8113 cells markedly inhibited the expression of the mature markers of DCs including HLA-DR, CD80, CD86, CD40 and CD1a, as well as the immature marker CD83, while inhibition of human VEGF in Tca8113 cells significantly reversed these effects. Therefore, human VEGF in Tca8113 cells may not only regulate the cell proliferation and apoptosis of oral squamous cell carcinoma cells, but may also inhibit DC maturation.
[Show abstract][Hide abstract] ABSTRACT: Successful treatment of cancer with dendritic cell tumor vaccine is highly dependent on how effectively the vaccine migrates into lymph nodes and activates T cells. In this study, a simple method was developed to trace migration of dendritic cells to lymph nodes.
Superparamagnetic iron oxide (SPIO) of γ-Fe(2)O(3) nanoparticles were prepared to label dendritic cells generated from bone marrow of enhanced green fluorescent protein (EGFP) transgenic mice, to explore the fluorescence intensity of EGFP influenced by the SPIO, and to make images of labeled dendritic cells with the help of magnetic resonance imaging in vitro. The SPIO-EGFP-labeled dendritic cells were injected into the footpads of five mice. After 48 hours, magnetic resonance imaging, optical imaging, confocal imaging, and Prussian blue staining were used to confirm migration of the SPIO-EGFP-labeled dendritic cells into draining lymph nodes.
The synthetic SPIO nanoparticles had a spherical shape and desirable superparamagnetism, and confocal imaging and Prussian blue staining showed perfect labeling efficiency as well. Furthermore, the dendritic cells dual-labeled by SPIO and EGFP could migrate into lymph nodes after footpad injection, and could be detected by both magnetic resonance imaging and optical imaging simultaneously, which was further confirmed by immunohistochemistry and Prussian blue staining. The percentage of dendritic cells migrated to the draining lymph nodes was about 4%.
Synthetic SPIO nanoparticles are strong contrast agents with good biocompatibility, and EGFP transgenic dendritic cells can be labeled efficiently by SPIO, which are suitable for further study of the migratory behavior and biodistribution of dendritic cells in vivo.
Full-text · Article · Oct 2011 · International Journal of Nanomedicine
[Show abstract][Hide abstract] ABSTRACT: To investigate the effect of vascular endothelial growth factor (VEGF) on the differentiation, formation and function of the dendritic cell (DC) in peripheral blood of patients with oral squamous cell carcinoma (OSCC).
Flow cytometry was used to detect the number of DC in peripheral blood of 81 patients with OSCC, and ELISA applied to test serum VEGF concentration the OSCC patients, and immunohistochemistry used to observe the expression of VEGF in primary foci of 57 patients with OSCC. DC from CD-14 peripheral blood mononuclear cells were cultured with VEGF(165) in vitro to investigate the cytokine's effect on DC.
In comparison with controls [(325.70 +/- 117.54) ng/L], the level of serum VEGF [(764.33 +/- 263.64) ng/L] was significantly increased (P < 0.01) and the DC numbers was significantly decreased (P < 0.01) in patients with OSCC. There was a negative correlation between serum VEGF concentration and the level of DC (P < 0.01). The expression of VEGF in primary focus was positively correlated with serum VEGF concentration, but was negatively correlated with the level of peripheral blood DC (P < 0.01). DC cultured in vitro with VEGF(165) decreased the expression of CD-1a, CD-40, CD-80, CD-86, CD-83, HLA-DR, and revealed a lower ability of stimulating T lymphocyte proliferation but a higher ability of uptake, compared to controls.
The overexpressed VEGF in patients with OSCC might be one of the important reasons for blocking the differentiation and maturation of DC.
No preview · Article · Apr 2009 · Zhonghua kou qiang yi xue za zhi = Zhonghua kouqiang yixue zazhi = Chinese journal of stomatology
[Show abstract][Hide abstract] ABSTRACT: With a unique electrochemical deposition method we fabricated in-plane arrays of straight cobalt filaments with periodic corrugations over a silicon substrate without using templates. The periodic corrugations on the filaments are induced by spontaneous oscillation in electrodeposition, and the periodicity can be tuned from a few tens of nanometers to a few hundreds of nanometers by controlling the electric current in experiments. Magnetic force microscopy indicates that each corrugated structure on the filament may correspond to a local single magnetic domain. When the inter-filament separation is large, the magnetic domains are regularly aligned along the filament. The domains become random when the filaments are closely packed. We suggest that our results could be helpful in understanding the evolution of magnetic domain patterns on microscopic scale and may have potential application in spintronics.
[Show abstract][Hide abstract] ABSTRACT: Arrays of straight cobalt filaments with periodic corrugation on the surface are electrodeposited on a glass substrate without using templates. The corrugated structures on the filaments are induced by the spontaneous oscillation of electric voltage in electrodeposition, and the periodicity can be tuned by applying different electric currents. Magnetic force microscopy indicates that when the interfilament separation is large, the magnetic domains are regularly aligned on the filament. Each corrugated structure on the filament surface corresponds to a local single magnetic domain. The domains become random when the interfilament separation becomes very small. We suggest that our results could be helpful in understanding the formation of magnetic domain patterns on a microscopic scale and may have potential application in spintronics.
No preview · Article · Jan 2009 · The Journal of Physical Chemistry C
[Show abstract][Hide abstract] ABSTRACT: The diagnosis of squamous cell carcinoma (SCC) of the tongue metastasising to the lingual lymph nodes is rare. Until now lingual lymph nodes have not been mentioned in any system of lymph node classification. We report a case of SCC of the tongue with metastases to the lymph nodes in the superficial floor of the mouth, and to levels I and II lymph nodes. We provide evidence of the existence of lingual lymph nodes, and validate the practice of removing the lymph node-bearing-tissue of the floor of the mouth during neck dissection in cancers of the oral cavity.
No preview · Article · Aug 2008 · British Journal of Oral and Maxillofacial Surgery
[Show abstract][Hide abstract] ABSTRACT: To investigate the inhibitory effect of dendritic cells (DCs) on the growth of the implanted tongue squamous cell carcinoma (SCC) tumors in nude mice.
Being induced from human peripheral blood monocytes (PBMCs) with rhGM-CSF, rhIL-4 and rhTNF-alpha, DCs were pulsed by Tca8113 cells lysates. Those DCs and T lymphocytes were co-cultured to induce specific cytotoxic T lymphocytes (CTLs). Immunotherapy was then performed after those DCs and CTLs were implanted into the tumor-bearing nude mice.
DCs were induced from PBMCs with multiple cytokines. Compared with the control groups, the growth of tumors was significantly inhibited in the group that had been implanted with DCs and CTLs, and tumor doubling time also increased markedly (P < 0.05).
DCs function normally after being induced from human PBMCs with multiple cytokines; DCs pulsed with tumor cell lysates and CTLs co-cultured with those DCs have a significant anti-tumor immune activity in nude mice.
No preview · Article · Mar 2006 · Zhonghua kou qiang yi xue za zhi = Zhonghua kouqiang yixue zazhi = Chinese journal of stomatology