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-inducedrat kidney cells(RK3E-ras). We observed tumor as early as 3 days after injection of RK3E-ras cells in subcutaneous ofSprague-Dawley rats. Tumor size and volume were increased exponentially for 2 weeks. The tail vein injected rats obtainedthe lethal infiltration in the lung within 2 weeks. This tumor animal model has great potential for studying cancer processesand short-term screening of variable cancer therapy strategy.

A novel indirubin analog, 5'-nitro-indirubinoxime inhibits cell proliferation and induces apoptosis against various humancancer cells. In this study, we performed the microarray analysis to identify genes differentially expressed in the KB oralsquamous 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 controlcells. 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 withapoptosis (3.5%) and cell cycle regulation (5.8%). Our application of microarray analysis on 5'-nitro-indirubinoxime treatedoral cancer cells allows the identification of candidate genes for providing novel insights into the indirubin mediated antitumoractivity.

The purpose of this study was to evaluate the role of Fas, Fas-L, and FAP-1 expression in the oral squamous cell carcinomasand ameloblastomas. For this study, 10 subjects diagnosed as squamous cell carcinoma and 8 subjects of ameloblastomareferred to the Dept. of Oral Pathology, School of Dentistry, Kyung Hee University, 5 subjects of normal oral mucosawithout any inflammatory changes were used as experimental, control groups respectively. All the tissues ; experimentaland control group were fixed in neutral formalin solution and embedded in paraffin, serial tissue section weremade 5㎛ in thickness and processed in the standard way for immunohistochemical method, using primary antibody againstFas, Fas-L, FAP-1, each was diluted at 1;100 followed by the super sensitive non- biotin horse radish peroxidase detectionsystem with DAB as chormogen, counterstained with Gill's hematoxylin stain method , mounted. And examined under thebiologic microscope with the criteria of -(no epithelial stain), +(weak or focal epithelial stain), ++(moderate or focal intensiveepithelial stain), +++(intense generalized epithelial staining) for the epithelial, and connective tissue component insquamous cell carcinomas , ameloblastomas and normal oral mucosa on each. In normal oral epithelium, negative reactionwas noted on the Fas . Fas-L stain, but on FAP-1 reaction, tumors cells with intense reaction on nuclei and cytoplasm ornegative reaction on nuclei with intense reaction on cytoplasm were admixed. On Fas, Fas-1 reaction, both tumor cells ofameloblastoma 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 theFas and Fas-L, but it could suggest that FAP-1 induce the development of tumors by means of inhibition of the apoptosis.

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 inflammationand 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 expressionrate of COX-2 in histiocytes of periapical lesions(97.4%) was significantly higher than that of dentigerouscysts(69.2%)(p=0.0032). The expression rate of VEGF was highest in plasma cells in the periapical lesions(69.4%) and indentigerous cysts(69.2%). COX-2 expression rate in endothelium and VEGF expression rate in epithelium were significantlyhigher in the periapical cysts than in the periapical granuloma and. VEGF expression rate in endothelium of the periapicallesions was increased in cases with resolving inflammation. For the periapical lesions, epithelial expression of COX-2 andVEGF showed a positive relationship(p=0.0301), and endothelial COX-2 expression revealed positive relationship with VEGFexpressions in epithelium(p=0.0008), in histiocytes(p=0.0136), and in endothelium(p=0.0439). According to these findings, asthe periapical lesion progressed from granuloma to cyst, the expressions of COX-2 and VEGF were increased. COX-2 expressionwas significantly correlated with VEGF expression, which suggests that COX-2 seems to be involved in the progressionof periapical lesion by inducing the expression of VEGF.

Angiosarcoma is an extremely rare sarcoma arising from oral mucosa. We report a case of angiosarcoma in the rightmaxillary gingiva causing excessive bleeding. The lesion exhibited typical histologic features of angiosarcoma, showinginfiltrative proliferation of polygonal endothelial cells and arborizing blood vessels. Tumor cells showed expression ofCD 31 and factor VIII, but no expression of CD 34 antigen. The patient expired due to severe bleeding in the oralcavity.

In order to investigate stresses of dental patients in dental clinic by dental-analogous stimulations, we selected 23women and 36 men who are students of dental college in D university. This experiment was performed to compare andanalyze the changes of Galvanic skin resistance and heart rate by dental-analogous stimulations. In paired t-test ofmale group's GSR average, there were significant differences between sound and touch, and among pain and otherstimulations. And in paired t-test of female group's GSR average, there were significant differences among pain andother stimulations. In paired t-test of male group's heart rate average, there were significant differences between visionand pain, and between touch and pain. And in paired t-test of female group's heart rate average, there were significantdifferences between smell and pain. In unpaired t-test of male and female group's GSR average, there were nosignificant differences in smell, sound, vision, touch, and pain. In unpaired t-test of male and female group's heartrate average, there were no significant differences in smell, sound, vision, touch, and pain. It seemed that stressesduring dental treatment could be changed by surrounding circumstances and sex distinction.

Immuno-MemBlot is a technique for detecting, analyzing, and identifying proteins, similar to the Western blot techniquebut differing in that protein samples are not separated electrophoretically but are spotted through circular orslot templates directly onto the membrane. Recently we developed a new Immuno-MemBlot (IMB) method applying immunoreactionsand coloring procedures directly in the wells of MemBlot apparatus, which were connected by canals toperform drainage for reagent application and buffer irrigation. This IMB method was designed to get theimmunoblotresults more rapidly and clearly than the previous immunoblot ones. This study is aimed to evaluate the analytical accuracyof IMB using different biological assay. In the sensitivity test of IMB the monoclonal antibody can clearly detectthe 30 ng (about 12 pM) of Mucocidin peptide (35 mer), and is also available to detect at least 10 ng (about 4 pM) ofMucocidin peptide (35 mer). The IMB was effective in the quantitative analysis of methothrexate (MTX) assay for cellularapoptosis. And more, this IMB is useful to screen large number of specific samples with ease and accuracy in ashort time. In the screenings for the presence of Mucocidin in saliva the quantitative comparison is conspicuous among48 persons depend on the different conditions ofgender, drinking and smoking habits, and oral diseases. Therefore, itis presumed that, even though the target proteins were partly degraded, a specific epitope can be detected if a monoclonalantibody was still reactive. Conclusively, these data suggest that the IMB can be useful in the primary qualitativeandquantitative analysis of proteins in various fluids, i.e., blood, saliva, tear, urine, etc.

The periodontal ligament (PDL) is that soft, specialized connective tissue situated between the cementum coveringthe root of the tooth and bone forming the socket wall. The PDL is a connective tissue particularly well adapted to itsprincipal function, supporting the teeth in their sockets and at the same time permitting them to withstand the considerableforce of mastication. During the life time, PDL is usually exposed to mechanical stress by mastication.However, little is known about the gene which is related to the mechanical stress in PDL. UNC-50 (PDLs22) was identifiedand isolated from D. melanogater and C. elegance. This gene was also regulated in sensory bristle for mechanotransductionin D. melanogaster. In this study, to uncover the relationship between UNC-50 and mechanical stress, we induced the mechanicalstress by medium displacement in cementoblast cell line. After mechanical stress induction UNC-50 expression wasanalyzed by RT-PCR, Real-time PCR, and western analysis. The expression of UNC-50 was increased after medium displacementof cementoblast in vitro. Collagen type I, type III, and osteonection mRNAs were also strongly expressed aftermechanical stress induction. The results of this study suggest that UNC-50 might responsible for molecular event in PDLinducing cementoblast under mechanical stress.

Oral squamous cell carcinoma is the 1st most common malignancy in oral and maxillofacial area. HPV 16 has beenstrongly linked to progression of cervical carcinoma. E6 and E7 as a small DNA virus encoding two major oncoproteinsof HPV 16 can act together to produce efficient immortalization of primary human epithelial cells. Thus it is importantto pursue the development of Immortalized human oral keratinocyte(IHOK) culture model which could be related to thepathogenesis of oral squamous cell carcinoma. If we establish IHOK transfected by E6/E7 genes, IHOK will be acceptedas a model system for HPV-linked oral carcinogenesis. The purpose of this study were to culture primary normal humanoral keratinocyte(NHOK), and to establish IHOK for studying oral carcinogenesis in the future. NHOK was primarilycultured under normal culture condition, and transformed into IHOK by transfection of E6/E7 genes. After 100 passagesdepend on Ca++ condition, cultured IHOK was confirmed by growth curve, cornified cell envelope measurement,TGase 1activity, mRNA detection, tumorogenecity and anchorage independence assay. After 100 passages, cultured IHOKshowed most basal cell and monolayer of polyhedral cells under 0.15mM Ca++, and small area of stratification andflattened epithelial cells with irregular border under 1.2mM Ca++. The cultured IHOK showed relatively resistantgrowth under high calcium condition. The E6/E7 mRNA was detected in cultured IHOK by RT-PCR. During the terminaldifferentiation in cultured IHOK, increased insoluble cornified cell envelope formation was accompanied with inductionof TGase 1 activity. But the cultured IHOK showed less CEM and TGase 1 activity than those of cultured NHOK.Cultured IHOK showed non-tumorogenecity, but slight anchorage independence. We had developed a technique totransform NHOK into IHOK by transfection of E6/E7 genes. Cultured IHOK was established as intermediate stage cellto study the pathogenesis of human oral squamous cell carcinoma.

Amino acid transporters are essential for the growth and proliferation in all living cells. Among the amino acidtransporters, the system L amino acid transporters are the major nutrient transport system responsible for theNa+-independent transport of neutral amino acids including several essential amino acids. The L-type amino acidtransporter 1 (LAT1) is over-expressed to support cell growth in malignant tumors. The double stranded RNA-mediatedRNA interference (RNAi) analysis can be in a wide variety of eukaryotes to induce the sequence-specific inhibition ofgene expression. In this study, we examined the effect of LAT1 short interfering RNA (siRNA) on cell growth usingsiRNA of LAT1 in the KB human oral squamous cell carcinoma. In the RT-PCR analysis and western blot analysis, thesiRNA of LAT1 inhibited expressions of LAT1 mRNA and protein. The uptake of [14C]L-leucine was inhibited by siRNAof LAT1. In the MTT assay, the siRNA of LAT1 inhibited the growth of the KB cells in the time-dependent manner,indicating that the growth inhibition of KB cell by the siRNA of LAT1 is induced by the blocking of neutral amino acidtransport mediated by LAT1. These results suggest that the transport of neutral amino acids including several essentialamino acids into the KB human oral squamous cell carcinoma is mediated mainly by LAT1. Further, the LAT1 would bea new target for the inhibition of cancer cell growth.

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