Cyclosporin A(CsA) as immunosuppressive drug is used to prevent immune reactions after organ transplant. And also It is reported that the effect of CsA on osteoblast differentiation has been controversial according to dosage. The purpose of this study was to examine the effect of various CsA concentrations on osteoblast differentiation. According to different concentration o f CsA, growth curve, apoptosis index MTT assay, ALP activation and osteocalcin secretion, in cultured NHost were analyzed. Treating osteoblasts with low concentrations of CsA increased growth rate, MTT assay activity, ALP activation and osteocalcin protein levels in a dose-dependent manner, while high concentration showed opposite results. Therefore, these results showed that low concentrations of CsA increased osteoblast differentiation, while high concentrations elicited an opposite response, showing inhibition of CsA on osteoblast differentiation. It suggested that different CsA concentrations might play in regulating NHost differentiation, and its specific activation of lower concentration will represent a viable anabolic therapy for bone resorption disease in future.

Tumor cell biological factors, such as urokinase plasminogen activator(uPA) and its inhibitor plasminogen activator inhibitor- 1(PAI-1) play a role in tumor invasion, metastasis, and proliferation. These factors in patients with primary oral squamous cell carcinoma(Oral SCC) will be evaluated and correlated with clinicopathologic variables. However, relatively rarely has been known in oral squamous cell carcinoma in vivo and in vitro study . The purpose of this study were to investigate the protein expression of uPA and PAI-1 in oral SCC cell lines cell line compared to NHOK and to study migration and adhesion assay. All the cell lines were cultured under KBM bullet kit at 37℃ in a 5% CO2 incubator. We studied a possible association between cytosolic uPA and PA-1 concentrations in oral SCC cell line compared to NHOK using an enzyme-linked immunoassay(ELISA). Cell adhesion and migration assay were done in all the cell l ines. In migration assay oral SCC cell lines were about 70 folds higher than NHOK. In adhesion assay oral SCC cell line were about 7-12 folds higher than NHOK. uPA cy tosolic concentrations was about 15-19 folds and PAI-1 was 3 to 4.5 folds than that of NHOK. Both uPA and PAI-1 concentrations were correlated with migration and adhesion assay. High cytosolic concentrations o f uPA and PAI-1 were correlated with migration and adhesion assay . It suggested that these markers might be specific for oral SCC cell line and these results would be contributed to treatment and prognosis of human oral squamous cell carcinoma.

The origin of squamous cell components in salivary gland tumor has been not yet clarified in detail. The squamous cell differentiation from adenocarcinoma has been reported in various carcinoma by HPV transfection in vitro. The adenocarcinoma cells adjacent to the squamous cell carcinoma components were positive for HPV. This is thought to indicate that after adenocarcinoma cells are transfected with HPV, they undergo morphological changes, and that squamous cell differentiation follows. The purpose of this study were to examine the effects of HPV-16 E6/E7 gene transfection into SGT cell line from human salivary gland adenocarcinoma, and to study the relation between the E6/E7 gene and squamous differentiation. Plasmid pBR322 containing HPV-16 was transfected into cultured SGT cell line using lipofectin method. Hygromycin was used as a selection marker. The presence of HPV E6/E7, transglutaminase 1, and involucrin mRNAs and protein in E6/E7 gene transfected cells was investigated by RT-PCR and immunoslot blot method. The apoptosis index was analysed by flow cytometry. The growth rate of E6/E7 gene transfected cells was reduced. E6/E7 transfected SGT cells increased apoptosis index. Involucrin and TGase I mRNAs by the squamous cell differentiation was most conspicuous in the E6/E7 gene transfected cell compared with non transfected cells. Squamous cell differentiation demonstrated in the transfectedSGT cell line, which expressed E6/E7 fusion gene mRNA.E6/E7 gene transfected cells showed squamous cell differentiation, expressing involucrin and TGase 1 protein by immunoslot blotting. The transfected SGT cell which expressed E6/E7 gene mRNA showed the squamous cell differentiation particularly clearly, and apoptosis was also demonstrated. It suggested that E6/E7 gene transfection into human salivary gland adenocarcinoma cells might induce clear squamous cell differentiation and contribute to study the pathogenesis of human salivary gland adenocarcinoma.

Urokinase-type plasminogen activator (uPA) and plasminogen activator type 1 (PAI-1) inhibitor contribute to the invasiveness of many carcinomas. It will be helpful to study clinical behavior of patients with malignant tumor by analysis of their expression. Expression of uPA and PAI-1 in human salivary gland tumors has been rarely reported in vitro. The purpose of this study were to investigate the protein expression of uPA and PAI-1 in SGT cell line compared to oral SCC and HeLa cell lines and to study migration and adhesion assay. All the cell lines were cultured under DMEM with 10% FBS at at 37oC in a 5% CO2 incubator. We studied a possible association between cytosolic uPA and PA-1 concentrations in SGT cell line compared to any other cell lines through cell migration and adhesion assay, and enzyme-linked immunoassay(ELISA). In migration assay SGT cell line was about 2 .5-4 folds higher than another cell lines. In adhesion assay SGT cell line was about 1.1-2 folds higher than another cell lines. uPA cytosolic concentrations of SGT cell line was about 3-10 folds, while PAI-1 was about 2.5-10 folds. Oral SCC cell lines were the lowest value. Both uPA and PAI-1 concentrations were correlated with migration and adhesion assay. High cytosolic concentrations of uPA and PAI-1 was correlated with migration and adhesion assay. It suggested that these markers might be specific marker for SGT cell line and would be contributed to treatment and prognosis of human salivary gland adenocarcinoma

Oral squamous cell carcinoma (SCC) is one of the leading causes of cancer mortality worldwide. It is generally thought that adjuvant chemotherapy provides modest prolongation of survival in various carcinoma. Docetaxel (Taxotere, TXT) play a significant role in the treatment of various solid tumors of epithelial origin. CsA (immunosuppressive drug) was widely used as adjunct for the treatment of cancer. Thus, it is important to pursue the apoptosis of IHOK and oral SCC induced by TXT combined with CsA related to the pathogenesis of oral SCC. But TXT combined CsA effect on IHOK and oral SCC remains unclear. After cultured IHOK and HN 22 oral squamous cell carcinoma cell line treated by 10 nM TXT and 1 μM, and caspase inhobitor, respectively, apoptosis index, cytochrome c and caspase-3 -8, -9 mRNA expression by RT-PCR, and procaspase-3 protein amount by immunoslot blotting was prepared. The purpose of this study were to examine the TXT-induced apoptosis pathway via caspase activation by CsA enhancement, and to apply these results to an effective therapeutic treatment plan for oral SCC by TXT combined CsA . 10 nM TXT showed about 60%, 55% celluar apoptosis of IHOK and HN 22, cell line, respectively, while CsA alone did not induce apoptosis in IHOK and HN 22 cell line. 1 μM CsA combined with 10 nM TXT increased apoptosis in IHOK and HN 22 cell line through caspase-3 and cytochrome c mRNA expression, while could not effect on caspase-8 and -9. Caspase inhibitor suppressed apoptosis of IHOK and HN 22 cell line induced by a combination of 1 μM CsA and 10 nM TXT. Immnoslot blotting showed procaspase-3 activation by a combination 1 μM CsA and 10 nM TXT, while caspase inhibitor inhibited activation. It suggested that a combination of CsA and TXT might induce increased apoptosis of IHOK and HN 22 oral squamous cell carcinoma cell line through caspase-3 activation. This treatment with a combination of TXT and CsA may be an effective therapeutic strategy for oral squamous cell carcinoma

The transcription factor nuclear factor-kB (NF-kB) plays an important role in regulating cell growth, apoptosis, and metastatic functions. Constitutive activation of NF-KB has been observed in various cancers; however, molecular mechanisms resulting in such activation remain elusive. Numerous evidences showed that over expression of TGase 2 might be linked with constitutive activation of NF-KB. To understand the pathways responsible for constitutive activation of NF-kB is important for rational design of NF-kB inhibitors for cancer therapy. Human salivary gland adenocarcinoma has been most aggressive solid tumors. Current therapy does not significantly improve survival rates. Thus, to investigate new therapeutic modalities against this adenocarcinoma is necessary. The purpose of this study was to study a constitutive activation of NF-kB with the expression of TG2 in SGT cell line origianted from human salivary gland adenocarcinoma by TGase 2 activity, RT-PCR and immunoslot blotting method. SGT cell line showed the highest TGase 2 activity and NF-kB activation than any other cell line. All the cell lines showed increased NF-kB mRNA activation after A231027 treatment than that of control. In immunoslot blotting, SGT cell line showed NF-kB activation correlated with TGase 2 expression after A231027 and BSA. It suggested that there might be a direct correlation between TGase 2 expression and NF-kB activation in SGT cell line.

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

Numerous bone cell culture models have been presented by the development of isolation and culturing techniques of cells. The culture of osteoblast-like cells of human origin with a specific osteoblastic phenotype has become an important experimental model in bone biology. Recently, it has become increasingly popular to utilize bone marrow cultures because these cultures are therefore thought to represent earlier stages of the osteoblast differentiation pathway. There is no report about culturing normal human osteoblast from oral and maxillofacial area. Primary cultured cells 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 from oral and maxillofacial cancellous bone and to analyze the phenotypic expression of cultured normal human osteoblast by the bone marrow isolation technique. Growth curve of NHost showed about 45hrs of doubling time and about 70μ g/well of total DNA content. NHost showed spindle shaped cytoplasm with ovoid nucleus under preconfluency and after cellular differentiation, they formed irregular numerous nodules from stratified cellular layers under D medium. ALP activity 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 under control medium. Scattered mineralized nodules stained by von Kossa method were seen on the cellular layer under D medium. It suggested that NHost might be established from oral and maxillofacial area by characteristic cellular shape, ALP activity, osteocalcin expression and numerous mineralized nodules.

It is not yet clear to know how normal human osteoblasts(NHost) from oral and maxillofacial area deposit, stabilize, and configure their extracellular matrix on dental biomaterial surfaces. Therefore it is necessary to design biomaterials with improved biocompatibility that will promote earlier bone differentiation and mineralization. There is now increasing evidence that TGase 2 may play a role in the initiation and regulation of the mineralization processes and serves to cross-link osteocalcin and osteopontin, which are predominant substrates for TGase 2 expressed during bone mineralization. But it is still unclear as to which TGase 2 is expressed by NHost in vitro bone formation. The purpose of this s tudy was t o determine the level of TGase 2 expression associated with collagen I , osteopontin and osteocalcin in NHost cell lines from oral and maxillofacial area in vitro. We will investigate whether TGase 2 has an active role in the biocompatibility of dental biomaterials during differentiation and mineralization of NHost. NHost cell lines were cultured under DMEM with 10% FBS at 37ﾟC and 5% CO2. Collagen quantification, mineralization and calcium assay, ALP activity assay, and RT-PCR analysis during bone differentiation and mineralization were done. ALP levels showed higher activity under AA+hGP t han under AA. I nhibition o f T Gase 2 by cystamine showed d ecreased Ca++ concentration, c ollagen I deposition and ALP level during 11 days of differentiation. TGase 2 mRNA expression of NHost was constant during mineralization stage. TGase 2 enzyme activity was increased during differentiation. Osteopontin mRNA expression during mineralization was higher than that of osteocalcin and collagen I . It suggested that TGase 2 associated with collagen I, osteocalcin and osteonectin might play a role in the differentiation of NHost from oral and maxillofacial area, but a little involved in mineralization.

Demineralized Freeze Dried Bone(DFDB) graft material have been used for reconstruction of large bony defects or augmentation of thin alveolar ridge during implantation of titanium fixtures. But at present osteogenic effect of DFDB do not overcome the capacity of autogenic bone graft. Thus many investigators had applicated various bioactive substance to DFDB to enhance the ability of osteogenesis of DFDB. In this study, mixture of grafting material was made from fibrin glue(F) and DFDB(D)(group 1: F+D), fibrin glue, DFDB and rhBMP-2(B) (group 2: F+D+B), fibrin glue, DFDB, polylactic- co-glycolic acid(PLGA)(P) and rhBMP-2(goup 3: F+D+B+P), fibrin glue, DFDB, PLGA, rhBMP-2 and autogenic osteoblasts( O)(group 4: F+D+B+P+O), and fibrin glue, DFDB, autogenic osteoblasts (group 5: F+D+O). During first surgical procedure, extraction of molar teeth was performed at male Biggle dog's mandible, and collected bone marrow tissue from tibia at same Biggle dog. Collected bone marrow tissue was cultured and differentiated into osteoblasts in vitro, and stored in nitrogen bottle. After four months, titanium fixture was implanted with prepared graft material to Biggle dog's mandible. After four and eight weeks respectively, experimental dog was sacrificed. Obtained tissues were prepared for examination by using resin embedded ground section method. Prepared sections were evaluated with transmitted and polarized microscope, and areas of osteoid and cacified bone were calculated with IPTK 5.0( image processing tool kit version 5.0). Resultant data was statistically analyzed by SPSS 13.0 software. Results of this study showed that autogenic osteoblats had more enhancing capacity of bone formation than rhBMP-2, but PLGA inhibited bone forming potential of bony tissue.

Taxol(paclitaxel) is used in chemotherapy against several cancer. Treatment of tumor cell lines with taxol induces apoptosis, but exact mechanism is not sufficiently understood. The purpose of this study was to investigate apoptosis by the inhibitory effect of paclitaxel on the motility properties of human salivary gland adenocarcinoma cell lines. Paclitaxel inhibited cell motility induced by soluble and immobilized attractant. It suggested that paclitaxel would be a potent inhibitor of salivary gland adenocarcinoma cell motility independent of its cytotoxic and apoptotic activity.

The prominent side effect of cyclosporin A, immunosuppressive agent, in oral tissues is gingival overgrowth(GO). It is characterized by the gingival enlargement with epithelial thickening, a large number of proliferating fibroblasts and overproduction of ECM components. Fibroblast accumulation in Cs A induced GO is caused by the Inhibition of apoptosis. But CsA effect on normal human oral keratinocyte(NHOK) remains unclear. Transglutaminase 2(TGase 2) which is expressed and activated in tissue where epithelial cells undergo apoptosis has been used as a marker for apoptotic cells. The purpose of this study were to study the effect of CsA on the proliferation and apoptosis of cultured NHOK by TGase 2 expression. After primary cultured NHOK was treated by 0.1, 1.0 and 10ug/ml Cs A, growth curve, MTT assay for succinyl dehydrogenase activity and RT-PCR for TGase 2 mRNA expression were done. The obtained results were as follows. MTT assay showed about 65% cell viability at 1.0μg/ml and 40% at 10μg/ml CsA. Growth curve showed normal S curve on control & DMSO, while decreased growth rate after 3 days of higher CsA tx. TGase 2 mRNA expression of cultured NHOK was the highest at 10μg/ml Cs A. TEM showed chromosome margination, and vacuole formation and clustered mitochodria in cytoplasm of cultured NHOK after CsA tx. It suggested that higher CsA might induce apoptosis of NHOK correlated with increased TGase 2 mRNA expression

Cyto kinc production by epiclerma l keratinocytes has been investiga ted ext ensive ly cluring the past decacle in the skm Furthermore‘ cyto kines produced by pidermal kerat inocytes may be regar decl as important regulators in inflammation, 1 nunune responses‘ and during wound healing, The associa tion of specific cytokine patterns in proliferative a ncl/or infl ammatory related cha nges in the s kin suggests a role in the pathogenesis of certain skin diseases, Although it is conceiva b1e that the pa ttern of cytokine express ion in o1'al kera tinocytes might be sirnilar t o tha t of epidermal origin, the1'e a re only sparse reports that have s hown t his experi menta lly, In addition, there is s ome evidence that there may be differe n ces in the proliferative capacity of oral versus epidermal keratinocytes, Since t his may be crucial for better un clerstanding the biologica l processes in the oral mucosa and how they may differ from the e pidennis, we a na lyzed the cyto kine expression pat tern of human oral kera tinocy tes, The purpose of this study were to investigate mRNA & protein express ion 0 1' va rious cy tokines between NHOK and NHEK by RT-PCR & immunoslot blotting, and to apply its results 1‘0 1' bet ter understancling t he pathological processes in the o1'al mucosal d1seases Cultured NHEK showecl larger a rea of cel lula r s tratil'icat ion tha n cul tu ++ 1'ed NHOK in 0 05mM Ca concentra tion, 1L - 1α , IL- 6 mRNA expression 0 1' cult ured NHOK we1'e hi gher than that of NHEK, TNF- mRNA expression of NHEK was about 1, 2 folds than that 0 1' NHOK, ICAM- 1 mRNA expression of NHOK was a bout 13 folds t han that of NHOK, while NHEK was weakly detected, 1L- 1a , IL-6 pro tein expression of cul tured NHOK were hi gher t han t ha t of NHEK TNF-a protein expression of NHEK was about 1, 2 fo lds than that of NHOK, 1CAM- 1 protein expression of NHOK was about 40 folcls than that of NHOK, whil e NHEK was weakly detectecl , mRNA express ion was associa ted wi th prot ein expression in cul tured NHOK ancl NHEK, It s uggestecl that lL- 1a ‘ 11-6 and ICAM-1 mRNA and protein be highl y expressecl in cultured NHOK, Especially, ICAM- 1 would be a useful ma rker for the pathogenesis of oral mucosal di sease,

AJthough salivary gland adenocarcinoma accounts for third prevalence rate of all salivary gland tumors. it is one of the most aggressive solid tumors. Current therapy does not s ignificantly improve survival rates. Thus‘ investigating new therape utic modali t ies aga inst sali va ry gland adenocarcinoma is necessary. Manumycin A. a natural product o{ Streptα7Jyces parvuJus‘ inhi bits farn esy l- transferase by competition with farnesyl pyrophosphate groups . Manumycin has shown antitumor activity in several ex per‘ imental systems through identifying the regulatory pathway of apoptosis. The hi erarchical relationship of caspase-8 to caspase-3 and caspase-9 in the drug-induced a poptosis pathway in antitumol effect is not clear. The hi erarchical relations hip between cytochrome c and the caspases and provided evidence to support the hypothesis that the release of cytochrome c was upstream of caspase activation in the enhanced apoptosis induced by manumycin A Manumycin A has not been examined extensively in human salivary gland tumor and has not yet been clarified. The purpose of this study were to investigate mRNA and protein expression of Bc l- 2 、Bax, Cytochrome C‘ caspase- 3 , 一8 and -9 in SGT cell line by RT-PCR and immunoslot blotting, and to a pply its results to exami ne iLs chemoprevention for salivary gland adenocarcinoma. MTI assay showed about 50% cellular viability of SGT cell line treated by 50μM manunycin A Bcl-2. Bax‘ and caspase-8 mRNA expression in SGT cell line were unchangeable after 50μM manu nycin A Cytochrome C‘ caspase-3 and -9 showed about 1.5-5 folds higher mRNA expression in SGT cell line than that of control a nd DMSO- t reated group a fter 50M manunycin A. Bcl-2, Bax, and caspase-8 protein expression in SGT ce ll line were unchangcable after 50μM manunycin A. Cy Lochrorne C, caspase-3 and -9 showed about 2-7 fo lds higher protein express ion in SGT cell line than that of control and DMSO-treated group after 50μM manunycin A. mRNA expression was assoc iated with protein expression in SGT cell line after 50μM manunycin A. It suggested that manumycin A would induce poptotic effect on SGT cell line by caspase-3 and - 9 activation through cytochrorne c release. And man umycin A will be a useful chemoprevention drug for human salivary gland carcinoma in future.

Nowdays, implant treatment became a branch of universal dental treatment and results in mOI‘ e success by surface con dit ioning and continuous improvements. Recently, a method to extract crystal shows sirnilar Ca/P ratio as HA has introduced which is anodizing electrolyte pure titanium anodize by electrolyte ß -glycerophosphate disodium salt hydrate and calcium acetic acid on pure t itanium before hydrothermal treatment ln t his study fixu res have divided in 3 group: Machined(Group 1), Anodized(Group 1I), implant whi ch has a surface containing Ca and P formed by anodization and hyd rothermal treatment( Group m). Total 18 fixtures were impla nted on rabbit which sacrifi ced on week 2 and week 4 for the histological specimens. By t hese specimens polarized microscopic view‘ ll1icro CT view‘ EPMA, ISQ value were ll1easured, cOll1pa red and a nalysed by each group to f igure out the possible clinical use of titanium implant containing Ca and P by hydrothe rll1al treatment and anodizing electrolyte. ISQ value had no s ignifïcance differences between each 3 groups, However in each group 4 , 8 weeks had hi gher ISQ value than 2 weeks 1n polarized rni croscope, calcification level was following Group 1I ‘ Group m, Group 1. 1n rnicro CT‘ formation of cancellous bone level was following ‘ Group 1I , Group m, Group 1. 1n EPMA, distribution and concentration of Ca was following : Group 1I was two t ill1es more higher than Group 1 , Group m. Group m were higher level t han Group 1. In distribution and concentration of P Group 1I was high er than the other group. but th e re were no statical s ignifi cances. Finally, anodi zed implant was the most exellent on the early bone for ll1a tion Containing Ca and P implant by anodizing and hydrotherma l treatment was more better than machined surface implant, but there is no effi cience at ear ly bone formati on

Many researchers are interested in wound healing in the t reatment of burns, prevention of post surgical adhesions and cosmetic s urgery by excess collagen production and scar formatlOn Synthetic epidermal substi tutes with cultured epi thelial cells seem to be an attractive strategy since keratinocytes have been demonstrated to modulate fibroblast growth and collagen synthesis. Bioa bsorbable and biocompatible chitosan structurally mimics hyaluronic acid. Recently, a bio compatible synthesi zecl ch itosa n-PVP(polyvinyl pyrrolidone) hydrogels demonstrated in vitro biocompat ibi li ty for bio medical applications . However. there is no re port on this hydrogeJ"s ability to modulate human gingival fibroblast growth. The purpose of this study were to investigate different growth modulation between human gingival fibroblast and normal human oral keratinocyte by chitosan- PVP hydrogel, and to apply this biocompatible synthetic polymer to oral and maxillofacial wound healing. We have synthesized a hydrogel from chitosan-PVP and examined its effect on human gingival fibroblast growth modulation in vitro. Non-toxic and biocompatible hydrogel with human gingival fi broblasts and epithelial cells was tested by MTT assay. HGF showed a higher growth proliferation than that of NHOK after cell seeding. In MTT assay, 30% hydrogel leach out products showed a higher cellular viability in NHOK than that of any other products. In MTT assay, 30% hyclrogel leach out products showed relatively lower cellular viability of HGF ln growth profile, NHOK showed about 7 fo lcls higher than HGF after 1 day, while about 2 fo lds higher after 5 days. And also NHOK showed above about 70% cell ular via bility from 1 to 7 days. It suggested that Chitosan-PVP hydrogel would inhibit relatively the growth of HGF and s timulate the growth of NHOK_ This phenomenon may prove to be of use in wound management 0 1' oral and maxillofacial area as epitheli al substitutes.

Cytokines play a vital role in the host immune response by regulating the development and function of im munocompetent ce11s One immunomodulatory agent that has received attention in oncology research recently is interleukin - lO(IL-lO). IL-IO inhibi ted tumor antigen presenta tion and induced energy in T lymphocytes that had been s timu lated by autologous MHC class II positive tumor ce11s Patients with head and neck cancer have been shown to exhibit profound irnmunosuppression. The mechani sm by which tumor ce11s alter immunological function in the host is poorly understood. Recently. production of biological active IL- IO was confirmed in ovar‘ian cancer, melanoma, skin cancel‘ & head and neck cancer, suggesting that IL- lO reduces the function of tumor infiltrating lymphocytes and contributes to the tumor growth. IL-IO expression has not been examined extensively in human oral cancer and has not yet been cla rified. The purpose of t his study were to investigate IL-IO mRNA and protein expression in NHOK, IHOK and oral squamous ce11 carcinoma(OSCC) ce11 line by RT-PCR and irnmunoslot blotting, and to apply its results to examine its thera peutic significance for oraJ cancers. Cultured NHOK showed a lower level of IL-IO mRNA and protein expression than cultured IHOK and HN 22 OSCC cell line under pre and postconfluency. HN 22 OSCC cell line under pre and postconfl u ency. showed the highest level of IL-I0 Cul tured IHOK showing a intermediate expression of IL- IO could be as a vaJ u a bJe marker for oral carci nogenesis ste p. During the terminal differentiation of a11 the ce11 lines, IL- IO ex pression was significantly unchangeabl e. IL- IO mRNA expression of a11 the ce11 lines was consistent with IL-10 protein expression. It suggested that IL- lO expression might play an important role in oral carcinogenesis and IHOK could be a valuable marker for oral carcinogenesis step. And aJso IL- 10 related gene may be future targets for gene discovery and possi bJy therapeutic intervention

Desmoplastic ameloblastoma(DA) is histologically characterized by extensive stromal collagenization or desmoplasia. ln this study, anti-cytokeratin 8/18, 13, 19 for pathogenesis as well as anti-PCNA for cellular proliferation, were used to det ect the expression of these proteins in the desmoplastic ameloblastoma Basal layers of tumor nest were negatively stained by CKl3, while suprabasal and inner cells were positive for CK13. CK8/18 and CK 19 was negatively stained in the peripheral portion of tumor nest in DA, whereas CK 8/18 was in central portion and CKl9 was positive in the su prabasal and some of central portion of the cel l nest. PCNA index of DA was 60 ::!: 14.6% to 95 ::!: 17 .2%. The peripheral tumor cells of the islets presented higher PCNA labeling index, while some cells in the central area of foll icle containing squamous like cells also presented negative PCNA labeling index. Especially tumor islands showed higher PCNA index than in main tumor mass. lt suggested that desmoplastic ameloblastoma might be composed of many different tumor cell types‘ and have hi gher pr이 ife r a ting activity in tumor islands of the desmoplastic stroma

This r esearch was designed to find the specific and economic methods of diagnosis about malignant melanoma For this study, we selected a typical case that was ambiguous in diagnosis between malignant melanoma and simple mela notic pigmentation, Tissue sections was st ained by H&E method, and immunohistochemical analyses was performed about 8-100 protein and MART-1 molecule, This research showed that MART-l had a more specificity for melanocytes than 8-100 protein , Patterns of MART-1 molecule distribution was more helpful in estimation of malignancies than 8-100 protein distribution patterns, On the basis of diagnostic usefulness and economical aspects, 8-100 protein and MART-1 molecule showed synergis tic and complementary relationship in confirming of tumor origin and they would be much useful for accurate diagnosis of malignant melanoma

Extensive oral mucosa loss from a variety of conditions is associated with significant functional morbidity and mortality. Although it is known that keratinocytes are a rich source of wound healing promoting factors such as transforming growth factor-β1(TGF-β1), it is not clear whether differentiated keratinocytes in a multi-layer form release this multi-functional growth factor. This study examined the hypothesis that keratinocytes in mono- and multi-layer forms expressed different levels of TGF-β1. When NHOK reached confluency in serum free medium(KBM), in test medium containing 1.2 mM Ca++ KBM NHOK were allowed to form multi-layers and differentiate. The purpose of this study were to investigate the mRNA level of TGF-β1, FGF-2, and TIMP-1 by RT-PCR analysis and also to evaluate the expression of TGF-β1 and involucrin in keratinocytes at different times of the onset of differentiation. The numbers and sizes of these nodules were increased as the process of keratinocyte differentiation proceed. Cultured NHOK in preconfluency under KBM medium expressed a significantly higher level of TGF-β1 relative to those grown in multi-layer forms, while the level of TGF-β1 mRNA gradually reduced to its lowest level at 7 days of growing cells in test medium. Cultured NHOK in preconfluency of KBM medium expressed a lower level of FGF-2 and TIMP-1 relative to those grown in multi-layer forms, while the level of FGF-2 and TIMP-1 mRNA showed the highest level at 3 days at gradually reduced to its lowest level at 7 days of growing cells in test medium. As a differentiation marker for keratinocytes at different time points, the highest level of involucrin mRNA expression was found at the later stage of cell differentiation. It suggested that the expression of TGF-β1 mRNA be consistent with the expression of FGF-2 and TIMP-1 mRNA in NHOK grown in high calcium medium during the terminal differentiation. But differentiated NHOK expressing higher involucrin mRNA could show constant espression of TGF-β1, FGF-2 and TIMP-1.