We have previously shown that 5’-nitro-indirubinoxime (5’-NIO) has potent anti-tumor effect in various humancancer cells. But the potential anti-invasive effect of 5’-NIO in salivary gland cancer has not been studied yet. Thegoal of this study is to evaluate the effect of 5'-NIO on salivary gland adenocarcinoma SGT cell adhesion and migrationto Type I collagen treatment. Western blot, adhesion and migration assay were performed to evaluate the impactsof 5’-NIO on the expression of MMP-2/-9 and its upstream signaling molecules after treatment of type І collagen.SGT cell adhesion to type I collagen is significantly suppressed by 5’-NIO. 5’-NIO decreased expression of β1 integrin,phosphorylation of FAK, MMP-2/ -9 compared with type I collagen treatment. In addition, 5’-NIO inhibitedthe migration of SGT cells treated with type I collagen. These results suggest that 5’-NIO could effectively inhibit theinvasion and migration of human SGT cells by downregulating the expression of β1 integrin and MMP-2/-9 and phosphorylationof FAK, Akt, and Erk. Adhesion and migration to type I collagen of SGT cells can be influenced through5’-NIO.

"The organotypic raft culture model has been shown to be a useful method for examining the effects of biochemicalmanipulation on various epithelial cell types, using in vitro conditions that simulate the in vivo environment of thetissue of origin. To investigate this method as a model for photodynamic therapy (PDT), we cultures the oral squamouscarcinoma cell line YD-10B on fibroblast-containing collagen gels at the air-liquid interface and assessed the efficacyof PDT using a photosensitiser 5-ALA-hexenyl ester. PDT effect on YD-10B organotypic raft cultures after twenty-four hours was determined. The number of residual cells was significantly reduced in comparison to control at allfour different conditions. PCNA immunostaining and TUNEL assay revealed that PDT inhibited cell proliferation andincreased apoptosis. These findings suggest that the organotypic raft culture model provides an effective and rapidlaboratory method of investigating strategies for PDT on oral precancerous lesions and squamous cell carcinomas. "

Cyclosporine A (Cs A) which is a highly lipophilic cyclic undecapeptide mainly used for its immunosuppressive propertiesexert a wide spectrum of biological activities including fungicidal antiproliferactive, anti-inflammatory and chemotherapueticeffects. Human salivary gland adenocarcinoma is very aggressive characteristics, which is need to getthe effective chemotherapuetic methods. Subconfluent SGT cell cultures have been treated with CsA at in vivo relevantconcentrations for 24h. MTT assay for cellular proliferation of cultured SGT cell line has been performed and TGase 1activity assay for cellular differentiation has been detected in the CsA-treated samples. It suggested that CsA couldhave an inhibitory effect in the proliferation of SGT cell line but no in the differentiation.

As the human mixed saliva plays important roles for the protection, regeneration, immunity, and molecular transfer/signaling in the oral and gastro-intestinal mucosa, the salivary contents have great implications for the generalhealth of human body. Nevertheless, the analysis method of human saliva has not been well developed up to date, becausethe proteins of mixed saliva are rapidly interacted with each other and easily degraded by proteolytic enzymesand microorganisms. This study aims to develop an immunoprecipitation-based high performance liquid chromatography(IP-HPLC) for the analysis of human mixed saliva. The representative IP-HPLC analyses were performed to compareamong different subjects in variable general conditions. Compared to the normal control the subjects suffered frombacterial infections of gastro-intestinal enteritis, chronic periodontitis, and acute necrotizing gingivo-stomatitisshowed dramatic increase of LL-37 level depending on the severity of diseases, while the subject suffered from Herpesstomatitis, a viral infection showed great increase of β-defensin 2. These data indicate that LL-37 in human mixedsaliva is more responsible to the bacterial infections of gastro-intestinal enteritis, chronic periodontitis, and acute necrotizinggingivo-stomatitis, while β-defensin 2 is more responsible to the viral infection of Herpes stomatitis. Thisstudy also suggeststhat the IP-HPLC be easily applicable to the wide range of biological samples for the quantitativeanalysis of an objective protein.

The human embryonic-lethal abnormal vision-like protein, HuR, stabilizes mRNA containing adenine- and uridine-rich elements in their 3’untranslated region. Because cyclooxygenase-2 (COX-2) mRNA is a cellular transcriptthat contains an adenine- and uridine-rich element, it can be regulated by the HuR protein. In this study, we examinedthe relationship between COX-2, HuR, MVD, and the clinicopathological parameters. Nineteen out of 43 cases ofHNSCC showed high level of COX-2expression, and 68% of these patients showed high COX-2 immuno-reactivity indicatingthe strong expression of the cytoplasmic HuR protein. Also, MVD expression in the cases with high COX-2expression was higher than in the cases with low COX-2 expression. These results suggest a strong correlation betweenthe overexpression of cytoplasmic HuR and COX-2 expression in HNSCC, and that COX-2 is associated withMVD in HNSCC. In conclusion, COX-2 regulated by cytoplasmic HuR may be a good tumor angiogenic factor inHNSCC.

Adenosquamous carcinoma (ASC) is a rare malignant epithelial tumor in the oral cavity. It is regarded as a variantof squamous cell carcinoma (SCC) because ASC originates from the surface epithelium, has similar clinical features,and includes SCC portion which is histologically identical to the traditional form. This case report deals with two casesof ASC occurring in the oral mucosa of old male patients. Both case has tubular structures lined by cuboidal, basaloid,or small round cells under the conventional SCC component, which was reminiscent of the glandular differentiation ofadenocarcinoma. On immunohistochemical studies, the glandular component of ASC showed positivity to CK-pan, CK7and HMWCK, while the dysplastic squamous cells exhibited negativity to CK7. Accordingly, CK7 as well as morphologicalfindings can lead towards differentiating ASC from other variants with gland-like structures, such as basaloidsquamous cell carcinoma and adenoid squamous cell carcinoma.

Primary localized amyloidosis of the palate is a rare condition. We present a case of localized amyloidosis of thepalate of a 49-year-old Korean female who suffered from asymptomatic nodular swelling. Microscopic examination revealedextensive deposition of pale eosinophilic amorphous material throught the lamina propria and within the wallsof small blood vessels. A diagnosis of localized amyloidosis was made after the biopsy revealed characteristic stainingon Congo red, and an extensive workup for systemic involvement was negative. Localized amyloidosis should be consideredin the differential diagnosis of palatal nodular lesions

Krox-25, a Kruppel type zinc finger protein, may play an important role for the morphogenesis of tooth in ectomesenchymalinteraction between enamel epithelium and odontogenic mesenchyme. The interrupted expression of Krox-25by antisense inhibition is supposed to affect the abnormal development of tooth germ similar to the deranged proliferationof odontogenic tumors. This study was performed to know the histomorphogenetic effect of Krox-25 antisenseinhibition in tooth germs of mouse embryos and to understand the abnormal expressions of Krox-25 in differentodontogenic tumors which proliferate in aberrant direction of ecto-mesenchymal interaction. Total 95 tooth germs obtainedfrom pregnant mice in the 13th day of fertilization were cultured with antisense oligonucleotides targetingmouse Krox-25 gene, and their histological patterns were compared with those of different odontogenic tumors, i.e.,ameloblastic fibro-odontoma (n=5), ameloblastoma (n=8), and ameloblastic carcinoma (n=2). Resultantly, the culturedtooth germs treated with antisense oligodeoxynucleotides produced primitive dentine and enamel by odontoblasts andameloblasts, respectively, but they aberrantly grew and formed abnormal tooth organs. Especially, the harmoniousgrowth of enamel and dentine formation was greatly deranged by the antisense inhibition in the organ culture system.These findings were much similar to the abnormal growth of odontogenic tumors. The relatively well differentiated enamelepithelium of ameloblastic fibro-odontoma showed irregularly strong reaction of Krox-25, while the poorly differentiatedenamel epithelium of ameloblastic carcinoma showed weak reaction. These data suggest that Krox-25 mayplay important roles for the histomorphogenesis of tooth germ by signaling the ecto-mesenchymal interaction betweenodontoblasts and ameloblasts in normal tooth germ development of mouse embryos as well as in cytodifferentiation ofodontogenic tumors.

Numerous bone cell culture models have been presented by the development of isolation and culturing techniques ofcells. The culture of osteoblast-like cells of human origin with a specific osteoblastic phenotype has become an importantexperimental model in bone biology. Recently, it has become increasingly popular to utilize bone marrow culturesbecause these cultures are therefore thought to represent earlier stages of the osteoblast differentiationpathway. There is no report about culturing normal human osteoblast from oral and maxillofacial area. Primary culturedcells from oral and maxillofacial cancellous bone were analyzed by morphologic features, total DNA contents,ALP, osteocalcin and von Kossa staining positivity. The purpose of this study were to culture the cell population fromoral and maxillofacial cancellous bone and to analyze the phenotypic expression of cultured normal human osteoblastby the bone marrow isolation technique. Growth curve of NHost showed about 45hrs of doubling time and about 70μg/well of total DNA content. NHost showed spindle shaped cytoplasm with ovoid nucleus under preconfluency and aftercellular differentiation, they formed irregular numerous nodules from stratified cellular layers under D medium. ALPactivity was about 2 folds higher under control medium with 10nM 1,25 (OH)2D3 than that under control medium.Osteocalcin expression was about seven folds higher under control medium with 100nM 1,25 (OH)2D3 than that undercontrol medium. Scattered mineralized nodules stained by von Kossa method were seen on the cellular layer under Dmedium. It suggested that NHost might be established from oral and maxillofacial area by characteristic cellular shape,ALP activity, osteocalcin expression and numerous mineralized nodules.

Epigallocatechin-3-gallate (EGCG) and theaflavins (TF) are polyphenols included in green and black teas,respectively. Both green and black teas have been studied for their potential health benefits for cancer. Hypoxia-induciblefactor (HIF) has been implicated multiple physiological and pathophysiological pathways, particularly,oncogenesis. But, the molecular pathways that govern the cell response to EGCG are not fully elucidated. The presentstudy investigated the intracellular mechanism in oral squamous cell carcinoma (OSCC) cells treated with EGCG, focusingon HIF-1 expression and its effect on epithelial phenotype. EGCG decreased phosphorylated Raf-1 protein in YD 8OSCC cell, but B-raf protein was not affected at all by EGCG and TF. In addition, we here found that EGCG regulatedHIF-1α expression independent of Raf-1 protein. Taken together with our previous result, the result imply that EGCGis attributed to the HIF-1α expression via Raf/MEK/ERK pathway, and the HIF-1α expression is associated with thechange of epithelial phenotype in OSCC cell.

Amino acid transporters play an important role in supplying organic nutrient to cells. The expression profile ofL-type amino acid transporter 1 (LAT1) and its subunit 4F2 heavy chain (4F2hc) on different differentiation stages in4-NQO induced rat tongue carcinogenesis was examined using immunohistochemical analysis. The gradually increasingLAT1 and 4F2hc expression detected during the multistep progressive change shows that the protein may have an importantrole i n the multistep tongue c arcinogenesis. Conclusively, LAT1 and 4F2 hc c an b e a useful b iomarker f or abetter understanding of multistep tongue carcinogenesis, while the specific inhibition o f LAT1 and 4F2 hc would be anew rationale for suppressing tumor cell growth in tongue cancer.

A patient complaining of severe pain in the right submandibular area showed a huge sialolith in radiogram. Duringthe operation, the submandibular gland was much indurated, and large amount of pus was discharged out at an incisionof the salivary gland. The removed salivary gland contained a huge sialolith in the major excretory duct of submandibulargland, which had an intact grayish-white surface in ovoid shape. In the histological examination its excretoryducts were extensively dilated without extravasation of saliva, and the involved salivary gland was almost destroyedby the granulomatous r eaction. Most of a cinar cells were d isappeared and r eplaced by ductal cells filled withexudative materials. The microsections of sialolith showed typical laminar structures of calcification containing amorphousbasophilic material in the center, in which a lot of Gram positive and Gram negative microorganisms werefound. In the center of sialolith numerous microorganisms were admixed with mucinous materials which were stronglypositive for the antibody of mucin-1, and formed multiple colonies. In the periphery of the bacterial colonies prolinerich proteins (PRPs) were condensely localized, and followed by the consistent positive reaction of transglutaminase 4(TGase-4). These data suggest that the sialolith of this study is formed from the primary nidus of bacterial colonyaggregated with salivary mucin-1 and PRPs by the crosslinking reaction of TGase-4.