1'he purpose 0 1' this study was to evaluate the ro1e of c• fos and c-jun expression in the salivary gland tumol‘ s , For this study‘ 11 s ubj ects of sali vary gland tumors ; 4 su이 ects of p1eomorphic adenomas, 3 s ubj ects of adenoid cystic car cinomas , 2 s ubjects of adenocar cinomas, 2 subjects of mucoepidermoid tumors, referred to the Dept, of Ora l Path College of Dentis t l'Y, Kyung Hee Univer sity, were used as experimental group, and 2 subj ect s of normal minor salivary gla nds without a ny infla mmator y changes, were used as control group respectively, All the tissues experimenta l and control group were fixed in 10% neutral formalin solution and embedded in paraffin , serial ti ssue section were made 5 I1I1l in thickness a nd processed in the s t andard way for immunohistochemical method, using primary and secondar y a ntibodi es, for c-fos, c-jun, foll owed by the Streptavidin-Horse Radish Peroxidase, all BioGenex U,S,A, made, appli cat ion counter s t ained with Mayer's hematoxylin stain method, mounted And examined under the biologic mi croscope, with the criteria : -(no epitheli al s ta in), +(weak or focal epithelial stain), ++(moder a te or focal intensive epithelial sta in)‘ +++(intense generali zed epithelial staining) for the epithelial, and stromal ti ssues on each Attained results as follows : 1 1n nonna l minor saliva ry gla nds , it is noted that negative responses on the acini minimal res ponses in nucl ei and cytoplams of se rous demilun e, myoepi theli al cells, and intensive r esponses in nuclei and cytoplam of ducta l cells to c-fos and c-jun, 2 1n the res ponses to c-fos, positive responses in nuclei and cytoplasms of the lining cells o[ the ad e nαd tissues, e pidermoid, and mucous cells in mucoepidermoid tumors are noted and in other tumor tissuses, negative nuclei wi th pos itive cyto plasms are revealed 3 1n the responses to c- jun, it is noted that positive r es ponse in nuclei and cytolasm i n the cells of adenoid tissues in pl eomorphic adenoma, epidermoid cell s, mucous cell in mucoe pi dermoid tumor but in other tumor s, only positive responses in cytoplasm are noted, Intensive r esponses on c-fos‘ c-jun were noted on the high a typical cells 1'his results suggest that c-fos and c-jun may be affected t o the reactivation on growt h and development of the salivary gland tumors
The number 01' patien ts with tongue carcinoma is increasing rapidly among young indiv idua ls in many parts of the worl d. Until now‘ most of studies were focused on the comparison malignancy wi th normal 01' dysplasia. There is little report of gene a lterations in normal to cancer , oral carcinogenesis. The purpose of present study is to evaluate the gene a ltc ration in every steps of oral carcinogenes is by DD- PCR. To induce tongue carcinoma in rat by 4- NQO. each dri nking water made 10ppm. 25ppm. 50ppm and control(o nly D.W without 4-NQO) . Specimens were classified into 4 groups s uch as co ntrol, I(mild & mocle rate dysplasia) , II(severe dysplasia and car cinoma in s itu) , III(carcinoma). Total RNA was ext racted and DD- PCR was performed using customized random primers. And to confïrmed t he results of DD-PCR‘ RT- PCR a nd real-time PCR with specific primers were carried out. There was phenotypic alteration in tongue 。f dosc a nd t imc dc pcndcnt man ncr. In gross examination, multiple papules, patch form or ulcerations were observed during 4 - NQO t reatment Hi s tologicall y, dysplasia was observed in 3 to 6 month and tumor formation in 6 to 8 month For DD-PCR, RT-PCR and real-time PCR, cyclophilin A, BAC RP23-372MB and BAC CH230-103E9 were differ entia lly expressed. Taken together, cyclophilin A has a role in all steps of oral carcinogenesis. BAC RP23-372N田is implica ted in carcinoma in s itu a nd BAC CH230-103E9 mRNA expression is assoicated with dysplasia and carcinom in s itu Conclus ively. some genes a re impli catcd a ll st eps of oral carci nogenesis, others are associated with one step, whi ch meant that genes are di fferentia lly expressed in every steps
Amino acids a re required fo1' protein synthesis and energy sources in all living cell s. System L amino acid trans porters neutral amino aci cls i n a Na +' - independent manner. In ma 1ignan t tumors the L-type amino acid t ranspor te r 1(LA1'1), the first isoform 0 1' system L. is highly expressed to suppor t tumo1' cell growth. ln the present study, the expression 0 1' the system L amino acid transporter and BCH cytotoxicity were examined and compa red in both rat gli a l and C6 g lioma cel ls 1'he rat glia l cells expressed the L-type arnino acid t ra nsporter 2(LA1'2). the second isoform 이 system L, with its subu ni t 4F2hc. whereas the LA1'1 was not expressed. 1'he C6 glioma cells expressed LA1'1 with 4F2hc but not LA1'2 1'he [14C]L-leucine uptakes by the glial and C6 glioma cells were inhibi ted by BCH in a co ncent ration-d e pendant man ner wi th the lC50 values of 270.0 :t 13.7 μ M and 73.1 :t 4.5 μ M, respectively. 1'he cell g rowth was inh ibi ted by BCH in the time, concen tration-dependant manner in the gli al and C6 glioma cell s . 1'he rate of cell growth inhi bit ion induced by BCH in the C6 glioma cells was hi gh er than that in the gli al cells 1'hese results s uggesL that the tra ns ports of neutra l amino acids inc1uding several essenti al amino acids into rat g lial and C6 glioma cel ls a re for the most part mediated by LA1'2 and LA1'1, respectively. Therefore‘ the rat glia1 and C6 gJioma cells are excell ent tools for examing the proper ties of LA1'2 and LA1'1, respectively. Moreover, the spec너 i c inh ibi tion 0 1' LATl in cancel cells might be a new rationa le f'or anti -cancer therapy
Agi ng adversely affects the structure and function of the saliva ry gla nd and leads to ma rked insulin resis t ance that correlates with red uced ins ulin s ignal t ra nsduction, lnsulin-like g rowth factors are k.nown to be regulator involved in embryonic a nd postnatal development with sequences 62% identical t o that of proins ulin , To determine whether i n suli n- like growth facto l' - l, -2, - 1 receptor a l'e involved in the changes in rat salival'y gland by aging, we a na lyzecl the qua nti tation of 19f - 1, -2, - 1 rece ptor mRNA in l'at salivary glancl between 2 weeks and 26 months after bir th using the competiti ve reverse t ra nscriptase-polymerase chain reaction(RT-PCR) methocl Between 2 weeks ancl 2 months age, sharp falls in the qua nt ities of Igf-2 mRNA were observed, W11ereas Igf- 1 receptor mRNA rose by aging, but not sig nifïcantly, The quanti t ies of 19f- l kept by aging, These change seem to be involvecl a role fol' the Igf-2 in sali va ry c1evelopment a ncl earl y growth is incli ca ted, Thus, t he dras tic changes in the qua ntities of Igf-2 mRNA in the ra t sali va ry gla nd by aging seem to be in volved in the development, early growth and homeostasis of sali vary gla nd,
We studi ed the difTerential elTect of vitamins A, C, U. and E on normal human 이 al keratinocyte(NHOK) , HPV-16 E6E7 immor talized human oral keratinocyte(1HOK) , Oncogene transfected HPV-16 immortalized ce1ls(OTOK) , and two ol'al sq ua mous cell line(HNSCC30‘ HNSCC31) according to carcinogenesis stage. The vitamin effect was evaluated by morphology. ce ll viabi lity. a nd orgnaotypic culture Vitamin A has a greater negative effect on growth for all NHOK IHOK HNSCC. es pec ially N-Ras t rans fected IHOK, Vitamin D & E revealed no significant cell activity on NHOK lHOK, ad OTOK Vitamin C was found increased cell viability to IHOK and OTOK 1n primary oral squmaous cell ca rcino ma (HN30 ). vitam in 0 and C showing increased cell growth , but vitamin E showing no effect 1n metastatic oral squamous cell ca rcinoma(HN31), vitamin C has prol iferative effect , but vitamin 0 & E has anti-proliferative effect Vitamin A t reated normal a nd ma lignant ce1ls by organotypic cu lt ure. showed reduction of epithelial layer and in vasion to connective tissue. , especia lly in 1HOK & oncogene-transfected 1HOK, 1n conclusion. three-dimensional culture sys tem may be useful as a model to acess the efficiency of agents such as a1l trans retinoic acid can preventing progression of these premaligant lesion to maligant oral carcinoma(ch emopreventive agent) .
It is well kwon that HPV have been strongly linked to progression of or al squamous cell carcinoma‘ Effici ent im mortalization of nonnal human oral keratinocyte(NHOK) should provid further evidence for the role HPV in tumorogenes is ‘ Because IHOK(I mmortali zed human oral keratinocyte) has been considered as a moclel syst em for study ing I-!PV- linkecl oral ca rcinogenes is , it is important to pursue the differenti ati al change of IHOK cul t ure moclel during t he culture passage, The purposes of this study were to examine the cha ra r’ ct eristic clifferential changes of cul turecl immorta lizecl human ora l keratinocytes during long term passage, and to apply these results to or al carcinogenes is in the future, NI-!OK was primarily incubated at 370C and 5% C02 under KBM bullet kJt IHOK was co ntinuously cul t ured towarcl 100th passage(two times per week) , Growth curve of NI-!OK and II-!OK clepend on clùture passage was taken For examining the cha racte ri s t ic clifferential changes of II-!OK, transrnission electron microscope, 1ì'ansgluta miase activity‘ E6/E7 mRNA detect ion, a ncl tumorogenecity were done 10th II-!OK showecl sl ight polygonal flattencl cells and sometimes apoptotic cells ‘ while 100th IHOK showecl increased polygonal cell s ‘ Cultu recl 100th IHOK showed r ela tively resis tant growth to high calcium than 10th II-!OK Microvilli from 10th II-!OK was not connect ecl with each other, ancl scatte red cytokeratin fil aments of 10th II-IOK. while decreased cytokeratin filaments in cytoplasm & prominent clesmosome of 100th IHOK. During the terminal differ entiation in cultured IHOK, induction of TGase 1 activity of 10th II-!OK was higher than that of100th IHOK mRNA E6E7 expresson was cletected and unchangable in both cul tured cells There was no tumorogenecity inclucecl by both culturecl cel ls. Although late passage IHOK showecl less r esemblance to NHOK, and lower TGase 1 acti vi ty than ea rly passage IHOK, it suggested that these cells should be 110t yet fully differ entiatecl to oral squ a mous ce ll carcinoma cells
Al thou gh calcifi cation is a common finding in inflammatory salivary gland disorders , saliva ry gland tumour ra rely s hows calcifications. A case of clear cell mucoepidermoid carcinoma(MEC) of the hard pa late with extensive intra tumoural calcifïcations vis ible on computed tomog r때hy(CT) scans and histologic sections is described. The calci fï caLion in the sali va ry gland tumour 0 1' the palate recogni zed by a CT scan s hould be considered in the differential diagnosis of a MEC The mechanism of the i ntratumoural calcifi cation in our case is speculated to be a result of a secretory fu nction 0 1' the tumour cells
The phylogeneticall y conserved nuclear factor 1 (NFI) gene fami ly encodes s ite-specific tra nscription factors essential for the development of a number of organ syst ems. There are four NFI genes in mamma ls (Nfi a , Nfib, Nfi c, and Nfix) and single NFI genes in Drosophila melanogaster, Caenorhabdi t is elegans, Anopheles spP. ‘ and other simpl e animals. It was reported that Nfia-defici ent mice exhi bit agenesis of the corpus call osum and other forebrain defects , wher eas Nfib-defi cient mice possess unique defects in lung ma turation and fo rebrain defect. Recently, it was also found that Nfic-defi cient mice exhibit agenesis of mo l ar서 roots and severe incisor defects. In the present study, we investigat ed the possible role of NFI-C in odon toblast diffe rent ia tion and root dentin formation using the innovative and invalua ble Nfic knockout mice model Nfi c-defi cient mice showed a berrant odontoblast differentiation and consequentl y abnormal dentin formation, while other t issues/organs in the body including ameloblasts of the enamel organ a ppeared to be unaffec ted and normal One of the most st r iking changes observed in these aberrant odontoblasts was t he absence of in tercellular junctions beLween them, r esulting in di ssociation of the cells and loss of th eir cellular polarity a nd organi zation. Surprisingly, these cells became trapped in dentin-like minerali zed t issue and thus their overa ll morphology r esembled osteoblasts and os t eocyt es. There was also an increased apoptotic activity in Nfic-deficient mice. These findings strongly s uggest ed that NFI -C plays a key role in odon tob last differentiation and survival in a cell type-specific manner.
조직 병리학 진단을 위하여 PA8, Masson-trichrome, von Gieson 등의 조직화학 염색 이나, PCNA, p53, 8-100 등의 면역 조직화학 염색이 널리 사용되고 있다 조직화학적 염색법은 포르말린 고정을 통히여 만든 현미경 절편 조직 내애 잔 존되어 있는 단백질, 지질, 당류 성분들을 각종 염색성 화학물질들과 반응시킨 결과로 생기 는 단순한 발색 정도를 현미 경 상에서 관찰하는 것 이며 면역조직화학적 염색은 특정단백질에 특이하게 결합하는 항체 를 이용하여 항원성 단백질의 존재 를 확인하는 방법이다 특히 면역조직화학 염색에서는 포르말린 고정과 에탄올 및 크실 렌 등의 처리 에 의하여 관찰 히고자 히-는 단백질이 번성 또는 변형되어서 헝체가 항원성 epito p e을 인지하지 못하는 경우에 는 관찰이 어럽다 따라서 연역조직화학 염색에 시용되 는 항체는 현미경 표본 처리에 의하여 변성 또는 변형되지 않는 항원성 epi tope 부분에 대 힌 헝체 를 사용하여야 한디. 이와 같은 방법으로 조직화학 염색이나 면역조직화학 염섹에서는 관찰히고자 하는 성분의 존재 여부를 확인할 수 있으나 아직도 그 성분의 량을 판별하여 비교하기에는 어려움이 많다 최근 생화학 및 분자생 물 학의 삐-른 발달에 의하여 각종 계 측 장비의 정확도가 높아지고 새로운 분야에서 첨단의 장비 들이 만들어 짐 에 따라 인 체의 신진대사뿐만 이니라 각종 생리 및 병리학적 기전들이 알려지게 되었다. 예 를 들면, 세 포 주기 (ceIl cycle) , 세 포 노화(senescenc e) , 세 포 지펼사(apoptosis) , 및 세 포 암화(ca r cinogenesi s) 등에 대한 cascade pat hway가 밝혀지 고 있 으며 이 를 직접 병리학적 진단에 용용하는 것 이 비람직하다 그러 나조직 병리학적 진단에 사용되는 현미경 표본은 일치직 으로 포르말린 고정에 의히여 단백질이 심하게 cross-linking 되고 에탄올, 크실렌 , 및 열처리 에 의하여 심하게 조직 이 변성되어서 현미경 절편을 사용하여 병인론을 밝히기 위한 단백질 분석이 거의 불가능한 상태이다. 이에 본 연구자들은 현미경 절편 표본에서 추출힌 단백질을 이용하여 정확한 단백질 정량분석이 가능한 dot blot 방법 을 개발하였기에 이 를 보고하는 바이다.
ASKl(apoptosis signal-regulating kinase 1) is an important mediator of a poptotic s ignaling initi ated by a variety of death stimuli, including tumor necrosis factor ‘ Fas activat ion, oxidative st ress, and DNA damage. It was originally discovered as a mitogen -activated protein kinase kinase kinase(MAP3K) with proapoptotic activ ity Wben the A8Kl is stimulated, it activates both the MKK4/MKK7-JNK pathway and the M와(3/MKK6- p38 ki nase pathway‘ leading to stress responses or apoptosis. Owing to its critical role in promoting apoptosis. ASKl activity is highly control led in cells by multiple mechanisrns, includ ing phos ph이 ylation , oligomeri zation, and protein protein inte ractions. Phosphorylation of A8Kl at Thr-845 has been correlated with its acti vation . while phos phorylation at 8er-83 by Akt/protein kinase B attenuates A8Kl activity . It has also been demonst rated that in tramol ecular interaction, probably between the NH2• terminal and COOH-terminal domains of ASKl, may be required to maintain A8Kl in its inactive state, whereas oligomeri zation of its COOH- terminal domains is correlated with ASKl activation. The most commonly observed means of ASKl regulation, however, is thrO\땅h protein-protein in teractions. Numerous proteins have been shown to bind ASKl to exert theiJ‘ reg버atory function. For example. binding of TRAF2 0 1' Daxx promotes ASK1 funct ion, whereas the kinase and proapoptotic activities of A8Kl a re inhibited by many other associated proteins, including reduced t hi oredoxin, glutaredoxin, Cdc25A‘ Hsp 72, A8Kl - in teracting protein 1. and 14-3-3 proteins. A novel mechan ism of the anti-apoptotic action of Raf-l via phys ical interaction with ASKl wi ll be described. Furthermore, signifi cance of phosphorylati on s ite of 8er-1034 in the C-terminal regula tory domain of A8Klin the apoptotic activity wil l be discussed
Iron has a I"ole not on ly in the sy nthesis of hemoglobin but also in cell growth, including tumor development and progression. Excess iron aids tumor development by catalyzin g t he production of oxygen radicals that may be proximate carcinogens and by being a limiting nutrient to the growth and repli cation of cancer cells . Iron chelators have been shown to inhibit the growth and/or induce thc apoptosis of malignant cell lines from leukemia‘ neuroblastoma, melanoma. hepatoma, Kaposi's sarcoma, and cervical cancer. To ow' knowledge, iron chelating agents man ifesting anti-oral cancer effects has not been reported so fa r, and there a re no comparative studies on the elTects of c1esferrioxamine(DFO) on skin keratinocytes vs oral keratinocytes and on imm o J떠li zed cells vs oral cancer cell s. We have found that the iron chelators, deferoxamine(DFO) exert potent time- and dose-dependent inhibitory effec ts on the growth of IHOK and HN4 cells. The major mechanism of growth inhibition after DFO t reatment was by induction of apoptosis, which is supported by AnnexinV-FITC staining. cell cycle analysi s‘ DNA laddering and Hochest staining. We reported that the ch elator st rongly activates p38 MAP kinase and ex tracellul ar signal - regul ated kinase(ERK) , but not activates c-Jun N-terminal kinase/stress-activated protein kin ase(JNK/8APK) . Interl eukin -8(IL-8) is an important cytokine involved in tumor growth and angiogenesis in a va ri ety of rnalignancies‘ bu t the regulation of IL-8 in oral can cer cells are understood. We investigated whether mi togen-activatecl protein kinases pathway is involved in iron chelator-mediated IL-8 proclllction in immorta li zecl a ncl ma lignant ora l keratinocytes. In this study we examined the role of p38 ancl extracellular s ignal- reglll atecl kinase-l/2 in t he expression of IL-8 by DFO.
자극성 섬유종은 구강 내 만성 자극에 의해 발생하는 증식성 질환 중 하니이다 만성적 자극 또는 손상 이후 일어나는 상 치 치유 기전은 상피 각화 세포, 섬유아세포 다형핵백혈구, 대식 세포 림프구‘ 비만 세포 둥이 작용하여 일어니는 복합적이 고 정교한 반응이다 이 중 대 식 세포와 비만 세포는 손상 직후 초기 상처 치유 기전에서 직용하며 혈관 형성 . 교원질 합성 증가. wound-breaking stre ngth 의 증가, 세 포외 기질 합성 둥에 관여하고 섬유아세포 유주‘ 증식 . 육이 조직 형성 . 교원질 합성 둥에 관여하는 성장 인자와 cytokine을 분비하여 상처 치유 과정에서 핵심적인 역할을 힌다‘ 봉 띤구는 초기 싱처 치유 기전에 작용하는 세 포 중 비민 세포와 대식 세포의 l말현을 통하여 자극성 섬유종의 발생 기전을 규명히고자 하였다 2001년 1월부터 2004년 12 월까지 연세대학교 치과대 학 구강병리학 교실에서 진단펀 자극성 섬유종 88예외 정싱 구강 짐믹 9예에서 toluicli ne-blue 엽색과 CD 68을 일차 항채로 사용힌 띤역조직화학적 염색올 시행하였다 또힌 지 극성 섬유종의 조직학적 소 견을 결절형, spread type, 과세포형 혈관 증식형 등 4 종류로 분류하였으며 각각의 조직 소견 분류형에서 비만 세 포와 대 식 세포의 발현 빈도와 상관관계를 SPSS(ver. 13.0) 을 사용하여 통계학적으로 분석하였다. 자극성 섬유종과 정상 점막군을 비 교하였을 때 자극성 섬유종에서 비만 세포와 대식 세 포의 발현 빈도가 유의하게 증가하였다(p (0.05) . 지극성 섬유종의 결 절형과 spreacl type‘ 결절형과 과세포형에서 비만 세포의 발현 비 율이 유의하게 증가하였으며, 대식 세포는 조직 소견 분류 형 간 통계학적으로 유의한 차이를 보이지 않았다. 비만 세포와 대 식 세포간 피어손 상관 계수는 과세포형에서 0.693으로 나 타났다(p=0 . 038) 연구 결과 싱처 치유 기전에서 작용하는 인자 중 비만 세 포와 대식 세포 발현 증가가 자극성 섬유종 형성 과정에 기여하는 것으로 생각되었디 비 만 세포와 대식 세포는 조직 소견 분류형에서 차이 를 나타내어 자극성 섬유종의 조직 학적 차이에도 기여하는 것으로 생각된다
Hemangiomas are different from true vascular malformations in thei l‘ pathogenesis and cl inical prognosis. There are sti ll no standardized antibodies to distinguish hemangioma and vascular malformation apparently. We compared juvenile hemangioma and vascular malformation with immunohistochemjstry using va ri OllS antibodies, i.e. , ANG, bFGF, VEGF. EGFR, vWF. PCNA. p53. maspin, and TNF- . A very st rong positive expression of ANG and vWF was observed mainly in the vascular endothelial cells of juvenile hemangioma. VEGF s howed st rong positive reaction in the juveni le hemangioma, but p53 showed no positive reaction. Ancl a strong positive reaction of ANG was observed in the vascular endothelial walls of vascular malformation. p53 was frequently positive in the lining endothel ial cells in the vascular malformatJOn Using a ntiboclies such as VEG F'. ANG. vWF which a re related to the proliferation and matllrity of the vessel components. and p53 antibodies in order to confirm between juvenile hemangioma and vascular malformation would be helpful
HuR(human embryonic-Iethal abnormal vi sion-like protein, ELAV)은 최근 염증반응 및 세 포성장 조절의 중요 기전 으로 관심 을 받고 있는 전사후 유진자 발한의 조절기전 에 관여한다 HuR은 3' -untranslated regi on에 AU- rich ele rn ents(ARE) 를 지닌 일부 mRNA들(에‘ c- fos, VEGF. COX• 2 동)의 안정화에 기여하여 mRNA의 증가 및 부가적인 딘백발 한을 증가시킨디 HuR 단백 은 이 러한 염증 및 세포성장의 생리적 기전 외에 종양발생과도 관련될 수 있으며 ‘ 유방암 난소 암 및 뇌 종잉 둥에서 f-luR의 발한증가가 보고된 바 있다 두경부 편평 세 포암종 전암성 및 양성 편평세포 병소를 대상으로 HuR의 발현양상을 띤역 조직회학적으로 살펴봄으로써 HuR의 발현이 두경부 펀평세포암종의 발생과 관련될 수 있는지 실펴 보고자 힌다 본 인 구는 80여1 의 두경부 편평세포암종 14 예의 전암성 편평세포병소 및 32 예의 양성 면평세포벙소를 실험대상 으로 이용히였다 두경 부 편평세 포암종은 AJCC(Amcrican Joint Comrnittee on Cancer) 의 분류법에 의한 TNM분류 , 병리 조 직학적 분화정 도 및 발생부위 별로 구분히였다 면역조직화학적 염색은 mouse anti-HuR monoclonal antibody(ZymeCl Clone 3A2) 와 Envis ion kit (DAKO) 를 이용하였다 염색결괴는 2명의 병리의사가 독립적으로 결과를 평가한 후 x 2 test fol' trends (SPSS‘ ve l'si o n12) 로 통계분석을 행하였다 HuR 염색반응을 핵과 세포질 부위로 구분하여 평가하고 비 교한 결과, 핵 과 세 포질 염색 결 괴 모두 편핑세 포암종 전암성병소. 양성병소에서 유의한 차이경향을 보였으며‘ 편평 세 포암종의 경우 임상벙 기 및 발생부위에 따른 유의 한 치이경향을 보였다 특히 , 후두에서 발생힌 편평세포암종은 세포질애서 강한 양성반응을 보였 다 본 연구결과, mRNA 안정화 인자인 HuR의 발현 및 세포 내 분포가 두경부 펀평세포암종에서 이상조절 됨 을 보였으며 그 이싱 조절 양상이 두정 부의 부위별로 차이가 있음을 확인하였다 따라서 향후에는 발현이상의 downstream effec ts 및 이} 후와의 관련성에 대힌 연 구기- 추가되어야 할 것은 여져진다
Ameloblastoma and adenomatoid odontogenic tumor showed quite different tumorigenesis and prognosis , Besides theil‘ growth potential and histological features , there must be an essential diffcJ'cncc in gene expJ'ession profile between ameloblatoma and adenomatoid odontogenic tumor , The gene expression profiles we1'e compared by im munohi stochemi stry and immunoblot methods using different monoclonal and monospecific antibodies against on cogenes, growth factors, signaling molecules‘ matrix proteins, enzymes, Based on the immunohi stochemical find ings previously J'epo1'ted in the literature we found some di stinguishing feature of gene expressions 1'0 1' the tu mOl'igenesis between ameloblastoma and adenomatoid odontogenic tumors , The hi s togeneti c and mol eculal' mechani sms of both tumors wiII be discussed
A novel indil‘ubin analog‘ 5’ nitro-indirubinoxime(Ol1) inhibits cell proliferation and induces apoptosis again st variolls hllman cancer cell s. ln this stlldy, we performed the microarray analysis to identify genes diffel'enti ally expressed in the KB oral sqllamollS carcinoma cells after treatment with 011 Of the 10‘ 800 genes a nalyzed , 1700 genes(15.7%) showed di fferent expression level in the 011-treated cells with respect to untreated control cel1s Arnong those‘ 263 genes(15, 5%) were down -reg띠 ated and 220 genes(12, 9%) were IIp-regulated more than 2-fold, Functionally related gene clllsters inclllde genes associated with signal transdllction(18, 1%) , especially genes re lated with a poptosis(3, 5%) and cell cycle reglllation(5. 8%) . Our application of microarray ar뻐ysis on 01l-treated 01'외 cancer cells al lows the identifi cati on of candidate genes for providing novel insights into the 011-mediated anti -tllmor actl Vl ty ,
Phosphatase and tens in homologue(PTEN) 은 phosphatidylinosi t이 3'-kinase에 대해서 빈대로 작용함으로써 세 포픽 애 있는 지질 잔류인 phosphatidyl i nosi tol (3 - 5) - tri - phophate(PIP-3) 를 탈인산화시켜서 세포증삭의 자극에 대 한 막수용체의 반응을 조절하는 종앙억 제유전자이다 CpG islan ds의 promoter 메틸화가 세 포주기 의 조젤 이나 DNA 복구외 관런된 유진자 기 능을 소실시키는 것으로 알려지고 있으며. 두경부 편평세포암종에서 p16INK4a, p14ARF, p15‘ D뻐-K, E-Cadhe rin‘ GST-P. hMLH1의 메틸화가 보고되어 있다 두경부 편평세포암종에서 10번 염색체의 소실 이나 장완의 번이기 관칠되었으며. 이러한 유전적 변이의 배경에 PTEN 유전자가 있다 본 연구는 두경부 편평세포암종에서의 PTEN의 떼틸회 빈도와 단백발한 을 알아보고자 하였다 Methylation-specific PCR(MSP) 로 파라핀 포매조직 ( 양성 싱피증식성 병 소 25 이l. 두경부 편평 세 포암 종 44 예) 과 동결절편조직(두경부 떤평 세포암종 4 예)에 대해 PTEN의 메틸화 빈도를 확인하였다 양성 상피 증식 성 벙 소 20 예. 두경부 편평세포암종 40예의 파라핀 포매조직을 이용하여 PTEN 단클론항체인 6H21을 이용하여 변역 조직화학 엽색을 시행 하였다 염색강도와 염색된 부위의 백분율을 곱하여 반정량적으로 점수화하였으며 조직학적 동급 벙기외 임상적 변수애 따 라 분류하여 연역염색의 정수를 비교하였다 양성 상피증식성 병소 25 예 중 13예에서 메틸화되지 않은 PTEN을 보였으니 nl1 틸 화된 예는 없었다 두경부 편평세포암종의 경우 파라핀 포매조직 44예 중 22 예에서 매 틸화되지 않은 PTEN 을 보였으니 때 틸화된 예는 없었으며 4예의 동결절편조직 중 한 예에서 PTEN promoter부위의 베 틸화를 보였다 양성 상피 증식 성 병소와 두경부 편평세포암종의 면역조직화학 염색 평균점수는 각각 69 1과 705 였다 편평세포암종의 경우 임싱 적 벙기 를 1기와 271 를 하나의 군(12 예. 평균점수 852) 으로 3기와 471 를 다른 군 (15예‘ 41 , 9) 으로 분류하여 비 교하았을 때 통제학적으로 유의한 차이 를 보였다(P=0 .017) 편평세포암종의 조직학적 분화도에 따라 고분화암종(15 예 87 , 0) 과 중둥도분회 및 저분화암종 (22 예. 61 . 6) 으로 분류하였을 때 분화가 좋지 않을 수록 평균점수가 감소하는 경향을 보였으나 통계혁적으로 유의힌 차이 를 보이지 않았다(P=O , 361)‘ 환자의 나이와 성별에 따른 차이는 없었다 본 연구결과, PTEN 단백 발현은 두경부 떤평 세 포임 종의 생 불 학적 행태 및 조직학적 둥급과 연관성이 있음을 시사한다. 두경부 편평세포암종 발임과정에서 prEN의 페 틸화는 관련성이 적 을 것으로 보이며, PTEN의 다른 유전적 맨이가 발암과정에 중요할 것으로 생각한다
Since oral keratinocytes represent the natmal target for HPV(human papill omavi ruses) infecti on, HPV infection may be involved il1 the developmel1t of oral SCC. Through compaJ'ing the morph이 ogic featw-es of NHOK to 다fOK accorcling to calcium concentration by TEM, immortali zed oral keratinocytes(IHOK) transfected by E6/E7 gene of HPV 16 have been gained wide acceptance as a model system for HPV-linkecl oral carcinogenesis. The purpose of this study were to exami ned the ultrastructural fcaturcs of culturcd NHOK, IHOK, and HN4 oral squamous cell CaJ‘CI noma celJ line, and to apply these results to oral carcinogenesis in the future, Prima:rily cul tlU'ed NHOK, IHOK ++ and HN 4 cell line which were cu ltu red under 015 and 12mM CaTT of 1ιBM bulJet kit For tra nsmission electronmi crosco py(TEM). under preconfluency‘ and after 3 days of postconfluency uncler 1.2mM Ca ++‘ cultured NHOK IHOK, and HN4 cell line were immediately fixed in 2, 0% gluta:raldehyde in O.lM cacodylate buffer(pH 7, 4) at 40C 1'01' 1h TEM of cultured NHOK under 1. 2mM Ca ++ showed increased tonofi laments‘ and vaculated ovoid cells wi th cornifi ed envelope, whi le cultured IHOK showed prominent microvilli , unilateral desmosome in microvillus‘ and tonof t!amen ts Under high calcium cu ltured IHOK showed less tonofilaments than that of cultured NHOK, while cu ltu red lHOK a nd HN 4 cell lines showed more increased desmosomes under high calclUm It suggested that the ultrast ru ctura l cha nges of cultured IHOK would be accepted as the morphologic changes of intermediate stage aJl10ng oral carci nogenesis ,
lndil‘ ubin is the active ingredient of Danggui Longhui Wan‘ a mixture of herbal medici l1e t hat is used to treat chronic myelocytic leukemia in tradi t ional Chinese medicine‘ He re, we show that new indirubin deri vatives 5' -ni tro-indirubinoxime, 5' -f1 uoro-indiru binoxime and 5’ -tri rnethylacetamino-i nd i 1'1.1 bi noxi me‘ have potent a n ti- proliferative activity on various human cancer cells and oncogenic RK3E-ras rat kidney cells with lC50 con centration ranging from 1-25 μ M, When the RK3E-ras cells were treated with indirubin derivatives for 24 h, the activity of caspase-3 and caspasc-7 was induccd followed by apoptosis , 011 the other hand, the activity of SAPK/JNK was inhi bited over the same period , lndirubin deri vatives a lso s howed strong ant i-tumor activity in rat s이 id and oral tumor models The inhibition of tlUl10r g:rowth was observed in animals beaJ‘ing RK3E-ras-induced turnor given subcutaneous dose of 100 mg/kg every other day for 10 days, Histologically, μeatment of indirubin derivatives caused significant inhibition of tumor formation associated wi th increased apoptosis and decreased proliferation of tumor cell s , These findings provide the potential value of indirubin derivatives as a novel candi date fOl 없l tJ -cancer agents ,
뻐ny studìes have shown the anti-proli ferative effects of irondeprivation on cancer cell s‘ but the effects 01' iron-chelators on oral cancer have not been clearly elucidated , To investigate the effects of an iron chelato r, desferrioxamine( 01"O).on the growth of ilIllTIortali zed human o1'al ke ratinocytes(IHOK), primary oraJ cancer cel ls(HN4)‘ metastatic oral cancer cell s(HN12) , and human skin keratìnocytes(HaCaT) in the MTr assay, three-dimensional(3D) raft cul tmes, Western blott ing, cell cycJ e analysis‘ nuclear staining‘ and cytochrome c expression for apoptosis s ig naling pathway were used OFO inhibited the growth of immortalized IHOK and HaCaT and mal ignant HN4 and HN12 keratinocytes in a time- and dose-dependent manner according to the MTT assay, The 3D organotypic cu l tu re also revealed that OF'O-treated cells showed less epithelial maturation, less surface keratinizati on‘ and de creased epithelia l thickness, The major mechanìsm of growth inhìbition with the micromolar 0 1"0 treatment was by the induction of apoptosis‘ which was supported by nuclear OAPI staining, ONA fragmentation analysis, and J10w cytometric analysis for sub-Gl phase ar rest and Annexin V-1"ITC stainìng, Furthermore‘ Bax expression in creased together with p53 and p21WAF1!CIPl, whìle the Bcl-2 expression decreased in the immortalized and malig nant keratinocytes treated with 01"0 , Time-dependent cytochrome c from mitochondria was observed in D1"O-treated [l-IOK and 0 1'머 cancel‘ ceJJ s, and was accompanied by the activation of caspase-3 in IHOK cells. These resu lts demonstrate that 0 1"0 has growth inhibitory effects on immortalized and malignant oral keratinocyLes Lhrough the induction of apoptosis and suggest that further evaluation of OFO as a potcntial thcrapcutic agent for human oral precancerous and cancerous lesions is warranted