Studies to evaluate distribution of markers in normal keratinocyte and their immortalized keratinocyte are appropriate to evaluate the normal and preneoplastic lesion of oral cancers as biochemical and cytochemical changes associate with tumorigenesis being not completely understood. Complementary DNA microarray containing 6000 sequence -verified cDNA elements was used to systematically characterize the variation in gene expression patterns of NHOK cells vs. immortalized keratinocyte by HPV16 E6-E7(IHOK). Examination of gene expression that is 85 clones cDNAs exhibits greater than 2 fold overexpression in NHOK probes relative to IHOK probe, 147 cDNAs reveal greater 2 fold overexpression in IHOK relative to NHOK probe.The high similarity in gene expression (96.5%) between IHOK and NHOK cells suggests that only an additional 232/6720 (3.5%) of the genome is differentially gene activated during HPV16 immoratlized keratinocyte growth and differentiation. Examination of gene expression that differs between NHOK and IHOK cellsapprear to be related to : cell adhesion & recognition, cell cycle regulator, apoptosis, transciption factors, growth factors and therir receptors, cytoskeletal and extracellular matrix proteins, signal transduction modulators and effectors, and miscellaneous. The gene expression of cell recognition factor such as endothelin 1, collagen IV, fibronectin, and SPR1 in IHOK were upregulated. Distinct or duplicated cDNA clones representing the same gene were typically clustered in adjacent rows in the clustered gene map. Therefore the differentially expressed and identified genes should be informative in studying oral epithelial cell carcinogenesis and such studies should foster the research of molecular markers allowing to assess the phenotypeof malignant epithelial tumor.

Metastatic spread to cervical lymph nodes(LNs) is a major determinant of outcome in oral squamous cell carcinoma (SCC). To provide an useful method for the detection of lymph node micrometastases, we fulfilled the histopathological examination and reverse transcriptase polymerase chain reaction(RT-PCR) using the paraffin-embedded LNs of oral SCC patients. In this study, 78 LNs from 12 patients with primary oral SCC were analyzed. Metastases in the regional LNs were evaluated by RT-PCR for squamous cell carcinoma antigen(SCCA) and cytokeratin 5(CK5). Detectability of metastatic LNs by RT-PCR was compared with histopathological examination. Of 78 LNs, CK5 and SCCA mRNA were detected in 32(41.0%) and 8(10.3%), respectively. Histopathologically, 10(12.8%) of 78 LNs were positive. CK5 mRNA was detected in all 10 histopathologically positive LNs. In contrast, SCCA mRNA was detected in 5 of 10 histopathologically positive LNs. These findings suggest that genetic diagnosis by RT-PCR based on CK5 mRNA expression may be sensitive and clinically useful technique to detect the presence of metastatic carcinoma cells in regional lymph nodes of oral SCC.

본 연구는 재래산양에서 복제 수정란의 생산효율을 향상시키기 위한 기초 자료를 제시하고자 체세포 핵이식을 실시하여 공핵세포의 종류, 핵이식란의 활성화 처리 방법 및 수핵난자의 조건이 체외발달율에 미치는 영향을 조사, 검토하여 핵이식란 생산을 위한 최적의 조건을 규명하고자 실시하였다. 공핵세포의 종류에 따른 핵이식란의 체외발달율은 융합이 이루어진 핵이식란의 활성화 처리 후 분할율은 귀 유래 섬유아세포를 공핵세포로 사용하였을 때가 로서 태아 유래 섬유아세포를

평균 연령 10.8세인 비만 아동 17명을 대상으로 12주간 영양교육을 중심으로 한 체중조절 프로그램을 실시한 결과 비만도와 BMI에서는 유의적인 변화를 보이지 않았으나 허리둘레 및 엉덩이 둘레가 유의적으로 감소하였고, HDL-콜레스테롤이 유의적으로 증가하였다. 지방 조직 분비 호르몬-resistin, adiponectin, leptin의 수준은 체중조절 프로그램 실시전과 후에 유의적인 차이를 보이지 않았다. Resistin과 leptin의 변화량은 BMI 변화량과 양의 관계를, adiponect의 변화량은 음의 관계를 나타내 체위와의 상관성을 보여주었으나 본 연구만으로 이들 호르몬들과 체위변화와의 상관성을 규명하기에는 부족했다. 또한 체중조절 프로그램 후 열량 외에 무기질과 비타민 등 대부분의 영양소 섭취가 감소하여 체중조절 프로그램을 진행할 때 열량섭취는 줄이면서 미량 영양소의 섭취는 유지할 수 있게 올바른 식품을 선택할 수 있도록 하는 교육이 강화되어야 할 것이다. 본 연구는 비교적 단기간의 체중조절프로그램의 효과를 살펴본 것으로서, 본 연구결과를 토대로 앞으로 영양교육의 내용을 수정하고, 체조성 변화에 대한 연구를 강화하며 좀 더 장기적인 체중조절 프로그램을 수행한다면 아동 비만 치료에 더 좋은 효과를 낼 수 있을 것이라 사료된다. 또한 비만과 체지방분비호르몬의 상관성에 대한 후속연구도 필요한 것으로 본다.

Cervical carcinoma is the 1st most common malignancy in korean females. HPV have been strongly linked to progression of cervical carcinoma. E6 and E7 as a small DNA virus encoding two major oncoproteins of HPV can act together to produce efficient immortalization of primary human epithelial cells, providing further evidence for the role of HPV in tumorogenesis. It is important to pursue the development of Immortalized human epithelial keratinocyte(IHEK) culture model which could be related to the pathogenesis between cervical and oral carcinoma. If we establish IHEK transfected by E6E7 gene, IHEK will be accepted as a model system for HPV-linked cervical carcinogenesis. The purpose of this study were to culture primarily normal human epithelial keratinocyte(NHEK), and to establish IHEK for applying these results to cervical and oral carcinogenesis in the future. The obtained results were as follows. 1. After 7-9 passages, cultured NHEK was almost senesce and disappeared, but cultured IHEK showed most basal cell and monolayer of polyhedral cells under 0.05mM Ca++, while small area of stratification and flattened epithelial cells with irregular border under 1.2mM Ca++. 2. The cultured IHEK showed relatively resistant growth to high calcium condition. 3. The mRNA E6E7 in cultured IHEK by RT-PCR was detected. 4. During the terminal defferentiation in cultured NHEK and IHEK, increase of insoluble cornified cell envelope formation was accompanied with induction of TGase 1 activity. But the cultured IHEK showed less CEM and TGase 1 activity than those of cultured NHEK. 5. Cultured IHEK showed non-tumorogenecity, but week anchorage independence. From the aboving results, we have developed technique to transform NHEK into IHEK by transfecting cells with E6E7 gene. Cultured IHEK was established as intermediate stage cell for studying the pathogenesis of human cervical carcinoma.

The purpose of this study was to evaluate the role of c-fos and c-jun expression in the oral squamous cell carcinoma and ameloblastomas. For this study, 12 subjects diagnosed as squamous cell carcinoma and 7 subjects of ameloblastomas referred to the Dept. of Oral Path. School of Dentistry, Kyung Hee University, 2 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 and secondary antibodies, for c-fos (Anti-c-fos protein, rabbit polyclonal kit at 1:100 dilution), c-jun( anti-c-jun protein, rabbit polyclonal at 1:100 dilution), all BioGenex U.S.A. made, followed by the Streptavidin - Horse Radish Peroxidase(InnoGenex, Human-Avidin kit) application, counter stained with Mayer's hematoxylin stain method, mounted. And examined under the biologic microscope, with the criteria-(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 mucosal epithelium on each. Attained results as follows ; 1. In oral mucosa, c-fos and c-jun intensely expressed on the all cell layers except on the basal layer. Intense reaction is noted in the c-jun than in the c-fos. and a few cells with positive cytoplasm, negative nuclear are scattered in all layer. 2. The response to c-fos in ameloblastomas, is various according to the histological type, but intense resposes are nodted in nuclear and cytoplasm on the tall columnar cells at the periphery of the follicles compare to that on the stellate cells. 3. The respone to c-fos in squamous cell carcinoma, intense reaction is noted in cytoplasm and nuclei of the tumor cells in well differentiated, poorly differentiated type. 4. The response to c-jun in ameloblastoma, it is noted that moderate respone in nuclear and cytoplasm, at the tall columnar cells at the periphery of follicular or plexiform type but intense respone was notes on the columnar cells, and stellate cell in cytoplasm and nuclear of acanthomatous type. 5. The respon to c-jun in squamous cell carcinoma, it is noted that intensive responses only in cytoplasm in well differentiated type, but intensive responses in nuclei and cytoplasm in the poorly differentiated type are revealed. Intensive responses on c-fos, c-jun were noted on the high atypical cells. This results suggest that c-fos and c-jun may be affected to the reactivation on growth and development of the squamous cell carcinoma.

Odontogenic ghost cell tumor (OGCT) is considered as a neoplastic counterpart of the calcifying odontogenic cyst (COC). β-catenin mutations have been described in COC suggesting a critical role in its histogenesis. In this study, we report a patient with OGCT contains a missense mutation on codon 3 (ACT -> TCT). Immunohistochemistry showed nuclear, cytoplasmic, and membranous accumulation of β-catenin in the tumor cells. TUNEL assay showed positive signals in nucleated cells adjacent to the ghost cells. Our data suggest that β-catenin plays an important role in the tumorigenesis of OGCT. OGCT may developed by improper differentiation process coordinated by WNT signaling pathway. Further studies are needed to determine a genotypic/phenotypic characteristics of ghost cell containing odontogenic lesions.

Ionizing radiation is a well- known therapy factor for human carcinoma cells. Genotoxic stress mediates cell cycle control, transcription and cellular signaling. In this work, we have used a microarray hybridization approach to characterize the cell type-

It is very little known that the molecular mechanisms control growth, cell differentiation, and invasion of ameloblastoma into bone. Tissue culture methods have also been used extensively for studies of the cell biology of ameloblastoma. The purpose of this study were to examined the ultrastructural features of ameloblastoma, and to apply these results to examine the pathogenesis of ameloblastoma in the future. Amelobalstoma was primarily cultured under 0.1, 0.15 and 1.2mM Ca++ of KBM bullet kit at 370C and 5% C02. For transmission electronmicroscopy(TEM), cultured ameloblastoma cells were immediately fixed in 2.0% glutaraldehyde in O.lM cacodylate buffer(pH 7.4) at 40C for 1h The ultrathin sections were stained with uranyl acetate and lead citrate, and examined by TEM. The obtained results were as follows. Primay culture ameloblastoma grown in 0.1 mM Ca++ showed interlacing papillary projections without desmosome within early passage(3-4). Primary culture amelobalstoma under high calcium showed prominent desmosomes or tight junctions within early passage. There was evidence of cellular degeneration, as exemplified by nuclear pyknosis, the margination and clumping of the chromatin, and vaculolation under high calcium. The sparse ribosomes, the cytoplasmic space filled with vacuoles, and the condensed mitochondria were seen under high calcium. From the aboving results, under high calcium primary culture ameloblastoma showed rapid cellular degeneration within early passage, indicating that the cells were gradually losing metabolíc actívitíes, leading to enventual cell death. It was thought that it would be necessary to establish cultured immortalized amelobalstoma cell line for studying the pathogenesis of odontogenic tumors.

The study of cornified cell envelope has been used to investigate the differentiation factors and to advance oral carcinogenesis, CE of human oral keratinocytes are in wet condition as saliva containing many proteases, growth factors, and many kinds of bacteria, The analysis of CE in Immortalized human oral keratinocyte(IHOK) derived from normal human oral keratinocyte(NHOK) will be used to study the pathogenesis of oral squamous cell carcinoma, The purpose of this study was to analyze the amino acid component derived from CE of cultured NHOK and IHOK, It will be helpful to study the role of transfected E6/E7 gene in forming CE, and to examine the pathogenesis of oral squamous cell carcinoma, After primry culture of NHOK, IHOK were cultured in KBM bullet kit at 370C under 95% C02 incubator, Growth curve according to calcium concentration, cornified cell envelope measurement(CEM), and protein chemistry for amino acid component of CE were done(Mena :f::SD) , respectively. The obtained results were as follows, lHOK showed small areas of stratification, more compact, with irregular border and tightly apposed cells in 1,2 mM Ca++, Cornified cell envelope exhibited an aggregated group of empty space surrounded by the remained cell membrane, During the terminal differentiation in cultured NHOK and IHOK, insoluble cornified cell envelope formation was increased, CEM of NHOK was about 4 folds than that of lHOK under high calcium, Amino acid component of both groups showed Pro/Glu(SPR) , Gln/Glu(lnvolucrin) , and Gly(Loricrin) in descending order, From the aboving results, ít was suggested that when the terminal dífferentiation in cultured NHOK and IHOK, major amino acid component of CE in cultured lHOK was the same to that of cultured NHOK, It was thought that E6 and E7 gene should be involved in preventing the differentiation and proliferation of IHOK from making CE,

In order to obtain novel genes related to the human craniofacial development, molecular cloning and sequencing, and in situ hybridization using craniofacial tissue sections were performed and followed by protein structure simulation. Totally 231 clones were obtained from the subtracted craniofacial tissue cDNA library of human embryo. Random cloning using the non-redundant clones from the craniofacial tissue of human embryo was done and obtained 398 clones from the premade human chondrocyte cDNA library. Their partial sequence data showed that 214 clones of subtracted cDNA library of craniofacial tissue were still non-redundant in Genebank search. And 20 clones among 498 clones of premade chondrocyte cDNA library were known to be undefined genes. Through in situ hybridization screening in the craniofacial tissue sections of 10 weeks old human embryo 36 clones were found to be positive in specific tissues. Depending on the cell types of sirnilar developmental origin, the positive reactions could be divided into five groups. Among the 20 clones of undefined genes from human chondrocyte cDNA library, 7 clones showed characteristic positive reaction in human cartilage tissue by in situ hybridization. From the simulated protein structure, motif analysis and in situ hybridization studies for the 7 undefined clones, Ch89, Ch96, Ch129, Ch285 clones may function in the outer space of the cell constituting a part of matrix protein complex, and Ch276 as a transmembrane protein which might partic ipate in matrix calcification around chondrocytes. Ch153 is a kind of antirnicrobial protein also acting as an inflammation mediator, and Ch334 clone is a zinc finger protein, of which expression increases in human adult tissues We presume these novel genes from human chondrocytes may provide a new path of chondrocyte development and functions of human craniofacial tissues

Nuclear factor 1 (NFI) was discovered as a protein required for adenovirus DNA replication in vitro, but it is now clear that NFI protein plays an important role in the expression of many cellular genes. NFI-C null mice demonstrated aberrant odontoblast differentiation, abnormal dentin formation, and thus molar lacking roots while other tissues/or gans in the body, including ameloblasts appear to be unaffected and normal. However, little is known about the mechanism of NFI -C function in odontoblast differentiation and dentin formation. In this study, in order to elucidate the molecular mechanisms of odntoblast differentiation, we examined morphological characteristics of the aberrant odontoblast in NFI-C null mice. we also evaluate the expression of dentin sialophosphoprotein (DSPP) and bone sialoprotein (BSP) mRNAs in the MDPC-23 cells by northern analysis after over-expression and inactiγation of NFI -C into mouse MDPC-23 cells Odontoblasts of the NFI-C null mouse were round in shape, lost their polarity, organized as a sheet of cells, and trapped in osteodentin-like mineralized tissue. Abnormal odontoblasts of NFI-C null mouse revealed the absence of an intercellular junctional complex known as the t erminal webs. MDPC-23 cells started to express DSPP mRNA beginning from the postnatal day of 14 and showed a steady increase as differentiating into odontoblasts. Over-expression of NFI -C increased the expression of DSPP mRNA. Inactivation of NFI - C induced BSP mRNA expression. These results suggest that NFI-C plays an important role in odontoblast differentiation in a cell-type specific manner and thus in dentin formation