Oral squamous cell carcinoma (OSCC) has been a focus of cancer prevention studies due to the fact that it occurs by a multistep process and that a precancerous lesion in the oral mucosa is easily accessible. The present study was aimed at developing an optical detection system using autofluorescence spectrum measurements for the early detection of oral cancer. The optical detection system was designed to use an excitation wavelength of 337 nm emanating from a Xenon lamp. Precancerous and cancerous lesions were created in the hamster buccal pouch by treatment with 7,12-dimethylbenz[a]anthracene (DMBA). Four groups of five hamsters each were used in this experiment. The right buccal pouch was treated with 0.5% DMBA to induce carcinogenesis while the left buccal pouch was treated with mineral oil as a control. The autofluorescence of both buccal pouches was measured weekly. A difference in the excitation pattern between the normal and the carcinogen-treated tissue was noticed after three weeks. Specifically, the intensity of the autofluorescence spectrum in the DMBA-treated buccal pouch was increased at wavelengths between 400 and 450 nm. The results of the autofluorescence measurements were compared to histological findings and show that the intensity of the autofluorescence increased along with the stage of epithelial dysplasia. Based on the fact that one of the autofluorophores in this tissue is NADH, we measured the fluorescence at the 450-nm NADH wavelength to conclude that the increased autofluorescence in the dysplastic areas may be caused by NADH. Based on these data, we suggest that autofluorescence optical methods are a useful tool for the early detection of oral cancer.

암 발생 이 후， 암세포외 섬유모세포는 cytokine. 성장인자 둥 분자를 통하여 상호 작용을 일으킨다 최근 cbemokines의 신호전달이 종양세 포와 기질 미세환경의 상호작용에서 암세포의 성장， 침윤， 이 동애 중요한 영향을 준다고 보고 되였다 본 연구에서 마이크로 어레 이 를 통하여 구강펀평 세 포암종 암세포외 구강편평 세 포암종 암조직에서 유래된 섬유모세포의 공동배양에서 섬유모세포 존재할 경우 구강 편평세포암종 세 포주애서 chemokine receptor를 포함한 다수의 유전자들이 발현이 증가되었으며， 또한 암세포가 존재할 경우 섬유아세 포에서 는 c h e mokines을 비 롯한 디수의 cytokine 유선자들의 발현이 증가 되였음을 알게 되었다. 암세포와 섬유모세포사이의 상호 작용 에서 영힘을 주는 chemokin않의 종류 및 역할을 진일보 분석 하였다 구강편평세포암종 (OSCC) 세포주 YD-lOB, YD• 38, HSC-2, HSC-3, Ca9-22 빛 구강편평 세 포암종에서 유래힌 섬유모새포 (cancer derived fibroblast; CF) 를 사용하였다， 마이크로 어레이와 re al- time PCR를 통하여 암세포외 CF의 공동배양에서 발현에 변화가 존재하는 chemokines과 cbemokine receptor를 분석하였고， Trans well assay와 wound hea ling assay을 시행하여 CCL7의 암세포 침윤에 미치는 역할을 검토하였고‘ F-actin 형광염색으로 암세 포 세포 골격의 액틴세사의 활성을 형태학적으로 관찰하였다 또한 ELTSA를 이용하여 CCL7 분비량 측정 및 발현대상을 확인하였으며 면역조직화희염색으로 구강편평세포암종 조직 에서 CCL7의 발현 을 조사하였다 암세포와 섬유모세포의 공동배양에서 CF 존재 시 암세포 에서 ch emokine recep tor를 포함한 다수의 유전자‘ CCR5. CEECAM1 동의 발헌이 증가되었으며. 또한 암세포와의 공동배양에 의해 CF 에서는 암세포가 존재 시 CCL7‘ CXCL1, CXCL3 등의 c h emokines의 발현이 증가 되었다 특히 HSC-2, YD-IOB‘ Ca9-22 OSCC 암세 포와 CF의 공동배양에서 CF의 CCL7의 발현이 증기히였다 Trans well assay와 wound healing assay 실험에서 CCL7를 처리한 OSCC 세포의 이 동은 농도 의존적으로 농도 증가에 따라 증가하였으며 a nti -CCL7에 의하여 억제 되었다 F-actin 형광염색에서 CCL7 침가에 의해 농도 의존적으로 암세포 세 포 골격 의 액틴세사의 활성을 나티내어 세포돌기가 증가도어 이동성 증가를 형태학적으로 확인하였다 CF와 구강암세포(YD- 10B， HSC-2. HSC-3‘ Ca922‘ YD- 38) 와의 공동 배양에서 CCL7 의 발현은 모두 증가 되었고 CCL7의 발현은 CF 에서 분비 되였음을 확인하였다 정상 치 은 조직 9건 OSCC 암환자의 조직 16건에 대하여 조직면역화학염색을 진행한 결과 암환자의 조 직애서 7 례 (43%) 에서 CCL7 말현 을 관칠하였다 구강편평 세 포암종 암세포의 침윤은 암세포 자극에 의해 섬유모세포가 활성화 되어 CCL7의 분비를 촉진하고 섬유모세포에 의하여 분 비된 CCL7은 암세포에 영향을 주어 암세포로 하여 금 세 포골격 의 변화를 일으키며 이는 세포이동을 촉진하여 암세포의 침윤을 유도함을 알 수 있었다‘

Matrix metalloproLeases(MMPs) a re a family of zinc dependent enzymes with the capacity to degrade extracellular matrix prote ins. MMPs express ion correlates with cancer cell invasiveness and metas taslS The purpose of our sωdy was t。 determine the role 0 1' stroma l fïbrob lasts in ca ncer cell invasi on by examining the expression and activity of MMP-2 and -9 We used a YD- lOB squalllous ca rcinoma cell line that was est a blished in Yonsei Univer s ity ‘ College 0 1' Dentistry. We then appli ed two types of s lrolllal fibroblastsdel‘ ived from normal gingival tissue and cancer supporting ti ss ue Morphologic examination of fïbroblas ls was carried out by rnicroscopy and doubling time was measured by MTI assay . YD-lOB cells were cultured by lh ree-d imensional culture using collagen gel with two types of flbroblasts To examine the stromal - mesenchyma l inleraclion. we used a direct co-cul tu re system between YD-IOB cells and fibroblasts. Gelatin zymography was performed to exa llline MMP-2 a ncl 9 activit ies. We found that cancer-derived fibroblasts di splay stel late-shaped cells wi th many cyLopl asmic processes. whereas normal gingi val fibroblasts were spindle shaped. The c1oubling time of bOLh lïbroblasLs was not statistically different. Three-dimensional co-culture 0 1' ca ncer cells with ca n cer- c1erived fïbroblasts induced the formation 0 1' multi - Iayered atypical cells, as compared to culturing with normal gin gival f lbrobl asts Both three-c1 imens ional culture ti ss ues inclucecl the invasive gro￦th of cancer cells into the dermal eq uival enLs . Gelatin zymography s howecl that gelatinolytic activity of MMP-2 was activaterl in both co-culture models. However , MMP-9 ac li vity was not a lterecl in YD-lOB carcinoma cells In conclusion. enhanced MMP-2 activity was inc1 uced by boLh cance r- c1eri vecl a ncl norlllal gingival fibroblasts. suggesting that the potential to invacle by stromal fibroblasts was noL l imilecl to ca ncer- c1 eri ved lïbroblasts

Epitheli a l mesenchymal interaction(EMl) is well known to be essential in eznbryonic deve]opment. wound hea]jng a nd ca rci nogenes is. Th is study was a i med to design in vi tro model for the investigation of protein analysis in epithe li al a ncl mesenchyma l i nteract ion(EMI) . This stucly usecl oral squamous cell carcinoma cell line(YD-lOB) . 1'0 investigate the clifference 0 1' protei n ex press ion of cancel‘ cells influencecl by variable in vitro conditions. three different models were des ig necl ; Collagen gel- basecl ca ncer cell culture model devoid of fibroblasts(C) , Direct coτulture moclel(M2) composed of ca ncer cells beneath co ll agen gel embeclded with Swiss 31'3 fibroblasts ‘ and Indi rect co-culture model(Ml) with collagen layer betwecn cancel‘ cells and collagen gel with f lbroblasts Two-dimensional electrophoresis was performed to compa re t he diffe r ence of protein express ion pattern of ca ncer cells aznong three znodel systems. Protein identification was done by MALDI-TOF. As res ults ‘ pl'O te in express ion pat tem of cancel' cells was quite different between znonolayer cul ture and coll agen gel based cultu re. Aclditiona ll y. protein expression was different between culture models with fi broblasts and without fibroblasts a ncl between ind irect contact and direct contact of two cell types ‘ Among differentia l prot ei n spots. catheps in D WäS iuenLifï ed by MALDl• TOF Cathepsin D exprcssion was increased from C model to 11띠 and M2 model by West em blott ing. suggest ing that cathe psi n D expression may be activated by direct and indirect stimulation of stromal fï broblas ts F' rom these resul ts ‘ these models could be appropriate for EMI study and cathepsin D mi ght be incluced by fi broblasts s timulation

Squamous cell carcinoma comprises about 95% of oral cancers. 까le gene디C 없mage in carcinogen-exposed fields is accumulated to transforrn norrnal mucosa in dysplas디c tissue and fmally invasive carcinoma through multistep process. This carcinogenic process has been a cause of the development of secondary tumors after the removal of primary carcmoma. πle improvement of therapeutic modalities of oral cancer has driven into the increase of multiple cancer occurrence in head and neck region. We experienced 3 pa디ents who had mul디ple squamous cell carcinomas in oral cavlty. π1ÎS study aimed to report multiple pr따laπ squamous cell carcinoma by clinical and pathologic examination and to discuss their molecular mechanism