Cyclooxygenase(COX)-2 is an enzyme engaged in the synthesis of prostaglandin(PG) and is upregulated by inflammation. It is related with the expression of the vascular endothelial growth factor(VEGF) in the angiogenesis of granulation tissue. By using immunohistochemical stains, the expressions for COX-2 and VEGF were evaluated according to the degree of inflammation and the type of lesion, and the correlation between the expressions were investigated in the 39 periapical lesions (7 periapical granulomas, 7 periapical granulomas with epithelium, 25 periapical cysts) and 13 dentigerous cysts. The expression rate of COX-2 in histiocytes of periapical lesions(97.4%) was significantly higher than that of dentigerous cysts(69.2%)(p=0.0032). The expression rate of VEGF was highest in plasma cells in the periapical lesions(69.4%) and in dentigerous cysts(69.2%). COX-2 expression rate in endothelium and VEGF expression rate in epithelium were significantly higher in the periapical cysts than in the periapical granuloma and. VEGF expression rate in endothelium of the periapical lesions was increased in cases with resolving inflammation. For the periapical lesions, epithelial expression of COX-2 and VEGF showed a positive relationship(p=0.0301), and endothelial COX-2 expression revealed positive relationship with VEGF expressions in epithelium(p=0.0008), in histiocytes(p=0.0136), and in endothelium(p=0.0439). According to these findings, as the periapical lesion progressed from granuloma to cyst, the expressions of COX-2 and VEGF were increased. COX-2 expression was significantly correlated with VEGF expression, which suggests that COX-2 seems to be involved in the progression of periapical lesion by inducing the expression of VEGF.
The purpose of this study was to evaluate the role of Fas, Fas-L, and FAP-1 expression in the oral squamous cell carcinomas and ameloblastomas. For this study, 10 subjects diagnosed as squamous cell carcinoma and 8 subjects of ameloblastoma referred to the Dept. of Oral Pathology, School of Dentistry, Kyung Hee University, 5 subjects of normal oral mucosa without any inflammatory changes were used as experimental, control groups respectively. All the tissues ; experimental and control group were fixed in neutral formalin solution and embedded in paraffin, serial tissue section were made 5㎛ in thickness and processed in the standard way for immunohistochemical method, using primary antibody against Fas, Fas-L, FAP-1, each was diluted at 1;100 followed by the super sensitive non- biotin horse radish peroxidase detection system with DAB as chormogen, counterstained with Gill's hematoxylin stain method , mounted. And examined under the biologic microscope with the criteria of -(no epithelial stain), +(weak or focal epithelial stain), ++(moderate or focal intensive epithelial stain), +++(intense generalized epithelial staining) for the epithelial, and connective tissue component in squamous cell carcinomas , ameloblastomas and normal oral mucosa on each. In normal oral epithelium, negative reaction was noted on the Fas . Fas-L stain, but on FAP-1 reaction, tumors cells with intense reaction on nuclei and cytoplasm or negative reaction on nuclei with intense reaction on cytoplasm were admixed. On Fas, Fas-1 reaction, both tumor cells of ameloblastoma and oral squamous cell carcinoma showed negative reaction on nuclei and cytoplasms. On FAP-1 reaction, tumor cells of oral squamous cell carcinomas showed more intensive response compare to that on ameloblastomas. Considering these results, the tumor cells of ameloblastoma and squamous cell carcinoma showed negative reaction on the Fas and Fas-L, but it could suggest that FAP-1 induce the development of tumors by means of inhibition of the apoptosis.
A novel indirubin analog, 5'-nitro-indirubinoxime inhibits cell proliferation and induces apoptosis against various human cancer cells. In this study, we performed the microarray analysis to identify genes differentially expressed in the KB oral squamous carcinoma cells after treated with 5'-nitro-indirubinoxime. Among the 10,800 genes analyzed, 1,701 genes (15.8%) showed statistically different expression level in the 5'-nitro-indirubinoxime treated cells with respect to untreated control cells. Among those, 263 genes (15.5%) were down-regulated and 220 genes (12.9%) were up-regulated more than 2-fold. Functionally related gene clusters include genes associated with signal transduction (18.1%), especially genes related with apoptosis (3.5%) and cell cycle regulation (5.8%). Our application of microarray analysis on 5'-nitro-indirubinoxime treated oral cancer cells allows the identification of candidate genes for providing novel insights into the indirubin mediated antitumor activity.
Several tumor animal models have been provided as a tool for developing cancer therapy. Here, we developed rapid, easy-to use, and cost-effective new rat animal model for invasion and metastasis of cancer using genetically k-ras-induced rat kidney cells(RK3E-ras). We observed tumor as early as 3 days after injection of RK3E-ras cells in subcutaneous of Sprague-Dawley rats. Tumor size and volume were increased exponentially for 2 weeks. The tail vein injected rats obtained the lethal infiltration in the lung within 2 weeks. This tumor animal model has great potential for studying cancer processes and short-term screening of variable cancer therapy strategy.
Adenocarcinoma NOS of salivary glands is characterized by a high rate of local recurrences and metastasis. Long-term survival rate of Adenocarcinoma NOS lis not promising. Thus, different chemotherapeutical approaches had been proposed for this neoplasm, including apoptosis induction by drugs. The current treatment of choice of adenocarcinoma NOS is controversible, and an effective treatment for them is not yet available. Chemotherpeutic agents that can be inhibit or reverse the tumor growth by targeting apoptotic pathways will be new candidates for cancer prevention and therapy. The purpose of this study were to study the effect of Brefeldin A(BFA) as apoptotic inducing agent in SGT cell line from human submandibular adenocarcinoma NOS and apply these results to make a plan of treatment and prognosis of salivary gland tumors involving adenocarcinoma NOS. SGT cells were treated with a 300μM BFA solution in serum-free medium during 18 hours. SGT cells were grown in DMEM with 10% fetal bovine serum served as controls. The growth curve and MTT assay for succinyl dehydrogenase activity were performed. For apoptotic analysis, fragmentation of genomic DNA was confirmed with gel electrophoresis. Transmission electron microscopy was assessed for the effect of BFA on SGT cells phenotype. Apoptotic cell recognition and counting were carried out with Annexin-V, caspase 3 and APo2.7 antibody through flow cytometry. Growth of SGT cell line was abrutply decreased after 1 day of BFA treatment. MTT assay for succinyl dehydrogenase activity of the cells showed about 55% after 300μM BFA treatment. Destruction of cellular organells, numerous vacuolation in the cytoplasm & nucleus, chromatin margination, & fragments of nucleus were seen with TEM after 300μM BFA treatment. DNA fragmentation of SGT cell line was induced by 300μM BFA treatment and confirmed by gel electrophoresis from genomic DNA extraction. Late apoptosis of the cells through flow cytometric analysis of Annexin-V staining as induced by 300μM BFA treatment. Early apoptosis of the cells through flow cytometric analysis of caspase 3 and Apo 2.7 staining was induced by 300μM BFA treatment. It suggested that early and late apoptosis of SGT cell line would be induced by Brefeldin A treatment in vitro study. This work evaluated the efficacy of BFA, a potent apoptosis inducer, on SGT cultured cell line. And BFA as chemotherapeutic agent will be used as the treatment choice for adenocarcinoam NOS, and be need to apply BFA to in vivo study & clinical approach in future.
It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatory effects of LED irradiation such as promotes stimulation of wound healing have been well known, there are few reports about molecular mechanism associated with cell cycle by LED irradiation. The purpose of present study was to examine the molecular event in cell cycle of LED irradiation on primary human gingival fibroblast(hGF) in vitro. The source of light for irradiation was a continuous-wave LED emitting at a wavelength of 635nm, and manufactured that energy density was 5mW/cm2 on sample surface. The hGF were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental samples were acquired at 0 (right after irradiation), 8 and 24 hour after irradiation. To investigate the molecular mechanisms associated with cell cycle, growth phase was determined by flow cytometry and mRNA expression of cyclin A, cyclin B, cyclin D1, cyclin E, cdc2, PCNA, p18, p27, p21, and p53 were determined by real time RT-PCR. Flow cytometric analysis demonstrated the percentage of cells in the G1 and S phase were decreased, but the G2 phase increased, which showed cells irradiated by LED were transitioned from S to G2 phase. For mRNA expression, cyclin B, cdc2, PCNA and p53 were increased at 0 hour after irradiation, and most of cell cycle molecules were increased at 8 hour after irradiation. At 24 hour after irradiation, cyclin A, cyclin E, PCNA and p18 were increased. Taken together, LED irradiation induced proliferation of hGF cells through transition from S to G2 phase.
Since ancient Eygypt, various dental materials were used for lost teeth including gold. The key point of this materials were nontoxic to human body. Since early of 1990's, dental implant was done for recovery of maxillofacial defects. From middle of 1970's, osseointergation concept of implant was introduced and performed in dental field. Biocompatibility of titanium showed good effect for osseointergration but had some problems (Galvance current and toxic corrosion) with suprastructures such as gold crowns. This study was performed to make safe dental implants which have reduced Galvanic currents and corrosion. 3 kind of dental casting gold alloys (different Gold contents, 1㎝×1㎝×0.1㎝ plates.) were used as experimental group, while Titanium were used as control group. Normal human osteoblasts(NHOsts)were cultured during 1-4weeks for histologic study. For analysing the calcium(Ca), Phosphorus(P) and alkaline phosphatase(ALP), NHosts were cultred during 2-23days. After experiments, histologic finding were observed by LSM and SEM. Ca, P, ALP concentration by automatic biochemical analyzer were analyzed by ANOVA test and linear regression method. The results were as follows. Biocompatibility of dental casting gold alloys were similar to titianium alloys histolgically. Biochemical analysis of dental casting gold alloys had no significant difference to titianium alloy except AIGIS-Fine. We could conclude that biocompatibility of dental casting gold alloys with high contents of gold were superior to that of low contents and alloys with high contents of gold had no significant difference from titanium on NHost culture. Gold dental implant might be better than titanium implant due to similar biocompatibility and no galvanic currency.