Adenocarcinoma NOS of salivary glands is characterized by a high rate of local recurrences and metastasis. Long-term survival rate of Adenocarcinoma NOS lis not promising. Thus, different chemotherapeutical approaches had been proposed for this neoplasm, including apoptosis induction by drugs. The current treatment of choice of adenocarcinoma NOS is controversible, and an effective treatment for them is not yet available. Chemotherpeutic agents that can be inhibit or reverse the tumor growth by targeting apoptotic pathways will be new candidates for cancer prevention and therapy. The purpose of this study were to study the effect of Brefeldin A(BFA) as apoptotic inducing agent in SGT cell line from human submandibular adenocarcinoma NOS and apply these results to make a plan of treatment and prognosis of salivary gland tumors involving adenocarcinoma NOS. SGT cells were treated with a 300μM BFA solution in serum-free medium during 18 hours. SGT cells were grown in DMEM with 10% fetal bovine serum served as controls. The growth curve and MTT assay for succinyl dehydrogenase activity were performed. For apoptotic analysis, fragmentation of genomic DNA was confirmed with gel electrophoresis. Transmission electron microscopy was assessed for the effect of BFA on SGT cells phenotype. Apoptotic cell recognition and counting were carried out with Annexin-V, caspase 3 and APo2.7 antibody through flow cytometry. Growth of SGT cell line was abrutply decreased after 1 day of BFA treatment. MTT assay for succinyl dehydrogenase activity of the cells showed about 55% after 300μM BFA treatment. Destruction of cellular organells, numerous vacuolation in the cytoplasm & nucleus, chromatin margination, & fragments of nucleus were seen with TEM after 300μM BFA treatment. DNA fragmentation of SGT cell line was induced by 300μM BFA treatment and confirmed by gel electrophoresis from genomic DNA extraction. Late apoptosis of the cells through flow cytometric analysis of Annexin-V staining as induced by 300μM BFA treatment. Early apoptosis of the cells through flow cytometric analysis of caspase 3 and Apo 2.7 staining was induced by 300μM BFA treatment. It suggested that early and late apoptosis of SGT cell line would be induced by Brefeldin A treatment in vitro study. This work evaluated the efficacy of BFA, a potent apoptosis inducer, on SGT cultured cell line. And BFA as chemotherapeutic agent will be used as the treatment choice for adenocarcinoam NOS, and be need to apply BFA to in vivo study & clinical approach in future.

The periodontal ligament (PDL) is that soft, specialized connective tissue situated between the cementum covering the root of the tooth and bone forming the socket wall. The PDL is a connective tissue particularly well adapted to its principal function, supporting the teeth in their sockets and at the same time permitting them to withstand the considerable force 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 identified and isolated from D. melanogater and C. elegance. This gene was also regulated in sensory bristle for mechanotransduction in D. melanogaster. In this study, to uncover the relationship between UNC-50 and mechanical stress, we induced the mechanical stress by medium displacement in cementoblast cell line. After mechanical stress induction UNC-50 expression was analyzed by RT-PCR, Real-time PCR, and western analysis. The expression of UNC-50 was increased after medium displacement of cementoblast in vitro. Collagen type I, type III, and osteonection mRNAs were also strongly expressed after mechanical stress induction. The results of this study suggest that UNC-50 might responsible for molecular event in PDL inducing cementoblast under mechanical stress.

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

Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) is a well-known inducer of apoptotic cell death in many tumor cells. 1RAIL is expressed in human placenta, and cytotrophoblast cells express 1RAIL receptors. However, the role of TRAIL in human placentas and cytotrophoblast cells is not. well understood. In this study a trophoblast cell line, JEG-3, was used as a model system to examine the effect of TRAIL. on key intracellular signaling pathways involved in the control of trophoblastic cell apoptosis and survival JEG-3 cells expressed receptors for 1RAIL, death receptor (DR) 4, DR5, decoy receptor (OcR) 1 and DeR2. Recombinant human TRAIL (rhTRAIL) did not have a cytotoxic effect determined by MIT assay and did not induce apoptotic cell death determined by poly-(ADP-ribose) polymerase cleavage assay. rhTRAIL induced a rapid and transient nuclear translocation of nuclear factor-kB(NF-kB) determined by immunoblotting using nuclear protein extracts. rhTRAIL rapidly activated extracellular signal-regulated protein kinase (ERK) 1/2 as determined by immnoblotting for phospho-ERK1/2. However, c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (p38MAPK) and Akt (protein kinase B) were not activated by rhTRAIL. The ability of 1RAIL to induce NF-kB and ERK1/2 suggests that interaction between TRAIL and its receptors may play an important role in trophoblast cell function during pregnancy.

In order to investigate whether or not CCD-980sk cell line can be affected by Korean Citrus junos and medicinal herbs, we examined the MTT assay when we treated Korean Citrus junos and medicinal herbs in CCD-986sk human fibroblast cell line. The samples t

Established SGT cell line from human submandibular gland adenocarcinoma was used to study the TGase expression on a cellular level in vitro. Transglutaminase 2(TGase 2) is assoacitated with apoptosis, GTP binding protein, and cell marix interaction. The role of TGase 2 in salivary gland tumors is not clear yet. The pupose of this study were to examine the TGase expression of SGT cell line compared to other tumor cell lines, and to apply these results to the pathogenesis of salivary gland tumor. TGase enzyme assay of SGT, SCC-15, HN 4 and HeLa tumor cell line was 3 times repeated, and calculated. Immunoslot blot for semiquantitative protein analysis was done. The obtained results were as follows. 1. SGT cell line showed the highest TGase 2 enzyme activity(about 6-16 folds) irrespective of pre or postconfluency. 2. HN 4 cell line showed the highest TGase 1 enzyme activity(about 2-3 folds) irrespective of pre or postconfluency. 3. Under postconfluency TGase 1 induction was not induced, but slightly increased in all tumor cell lines. 4. TGase enzyme activity in all tumor cell lines was accompanied with TGase protein formation. From the aboving results, the higher TGase 2 expression of SGT cell line suggested that they would come from submandibular ductal cells and have a important role in the pathogensis of salivary gland tumors.

본 연구는 안과에서 주로 사용되는 생체 염색시약으로 Fluorescein, Rose Bengal, Erythrosin B, Lissanúne Green B 올 인간의 결박세포인 Clone1-5c-4 cell line올 배 양하여 cytotoxicity릎 조사하기 위하여 1%, 0.25% 의 농도보 5분， 15분 동안 처리하 여 MTT assay, SRB assay를 실시하였다. 각 염색사약의 세포독성올 바교 검정한 결과， Rose Bengal과 Lissamine Green B에서 서I 포 증식 저해용이 30% 이상 나타났 고， 회복효과를 보기 위하여 시약 제거 후 1 일 배양하여 Ml‘T. SRB assay를 실시한 결과는 Rose Bengal에서 거의 비슷한 수준이었으나 Lissamine Green B 에서는 더 증 가하였다. 또한 5, 15분 동안 시약처리 직 후 subculture활 실시한 후 세포의 부착과 중식정도를 관찰하였고， 회복효과를 보기 위해 3 잊 후에 세포의 형태를 관찰한 결과， Rose Bengal은 거의 차이 가 없었으나 Fluorescein 에서는 회복효과를 보였다.0.25% trypsin을 처리하여 세포에 영향을 주어 세포의 인위적인 손상올 형태학적으로 비교 한 염색상 세포 저해 정도는 Rose Bengal 이 가장 높-았고， Fluorescein 이 제일 낮았 다. 이러한 결과로 볼 때 일부 생체 염색시약이 인간의 결막세포인 Clone1•5c -4 세 포증식을 저해하는 것으로 사료된다.

콘태트렌즈 다목적 용액 내에는 보존제， 소독제， 세척제， 윤활제 둥이 포함되어 있 으며 콘택트렌즈 표면에 침착하는 단백질올 제거하기 위해 물질의 계변에 홉착되어 그 표면 장력을 현저하게 저하샤키고 수액 내에 열정 농도이상으로 폰재할 때 미젤 구조를 형성하여 정상적으로 용해되지 않는 물질을 미첼구조내에서 용해시키는 특성 올 갖판 계면활성제를 사용한다. 본 연구논 계면활성제의 세척 효과와 세포독성 판계를 연구하고자 소프트 콘택트렌즈 의 세척용으로 사용되는 여러 가지 계변활성제 (Tyloxapol， Tromethamine, Poloxamine, Poloxamer40η) 가 생쥐의 섬유모세포인 L• 929 cell line에 미 치찬 저해 효과를 알아보 았다. 계면활성제를 농도별 <0,001-10%, w/w) 토 처리한 후 시간별 (24h， 48h)로 MTT 와 LDH 분석 방법올 사용하여 측정하였다. 단백칠 제거효과는 친수성 콘택트 렌즈 FDA공인 W 그룹의 렌즈 etafilcon A (함수융 58%) 를 인공누액에 65시간 담근 후 꺼내어 계면활성제의 농도와 처리시간이2， 24h) 에 따라 단백절 제거효과를 조사하였 는데 단백질이 침착된 친수성 콘택트렌즈가 각각의 계면활성제에 의해 제거된 단백질 량의 정량은 Bradford 정량 방법올 이용하여 측정하였다. 세포증식 저해효과는 24시간 처리결과 Poloxamine의 ;성우 0.1%이하에서. Poloxamer407은 5% 이하에서. Tyloxapol 은 0.1%이하에서， Tromethamine은 1% 이하의 농도에서는 세포독성윷 나타내지 않았 으나， Poloxamine의 경우 1% 에서， poloxamer407은 7% 에서， Tyloxapol은 1% 에서， Tromethamine은 1.5%에서 세포독성을 나타내었다. 세포소지판을 중심으로 평가된 MTT 분석 방법은 LDH 유리 분석 방법보다 민감하게 나타났다. 계면활성제의 세포독성이 없는 농도에서 단백질 제거효과눈 Tylox따Xll > Poloxamer-때7> Tromethamine > Poloxamine 순으로 콘택트렌즈에 침착된 단백질 제거량이 높았으며， 농도 의존적으로 효과가 높은 것으로 조사되었다. 이러한 결파똘 통해 미래의 계면활 성제는 세포독성이 낮고 세척효과가 뛰어나야 할 것으로 사료된다.

본 연구는 콘택트렌즈의 세척용으로 사용되는 여러 가지 계면활성제중 널리 사용 되고 있는 Tyloxapol과 Tromethamine의 단백질 게거 효과와 L-929세포에 미치는 저해효과를 서로 비교하기 위하기 위해 먼저 렌즈를 인공누액에 65시간 담근 후 계 면활성제의 처리시간에 따라 단백질 제거효과를 UV Photo-spectrometer를 이용하여 측정하였다. 또한 계면활성제의 L-929세포에 미치는 저해효과는 MTT assay, LDH assay 방법으로 측정하였다. 연구 결과 계면활성제가 L-929세포에 미치는 저해효과는 통일 농도에서 Tyloxapol 이 Tromethamine보다 높은 것으로 나타냈으며， 세포 수도 농도 의폰적으로 감 소하였다. 독성의 차이는 LDH assay 보다 MTT assay가 더 민감도를 보였다. 침착 된 단백질에 대한 세척효과는 동일농도에서 Tyloxapol 이 Tromethamine보다 뛰어났 다. 단백질 제거효과는 농도 의존적으로 효과가 높은 것으로 조사되었으며， Tyloxapol과 Tromethamine의 세포증식 저해를 나타내는 최저농도에서 Tyloxapol 이 Tromethamine보다 단백칠 제거효과는 월둥히 뛰어났다.