Indirubin is the active ingredient 0 1' a traditional Chinese herbal medicine, Danggui Longhui Wan, used for the t reatment of chronic myelocytic leu kelma The author reports that novel indirubin derivative, 5'-nitro-indirubinox ime (5'-NIO) , has potent a nti -proliferative activity on human o1'al cancer cells , Treatment of KB cells with 5’ NIO s howed a s t rong cell growth inhibit ion during indicated time point , A typical apoptosis pattern was obtained with DNA fragmenta tion and i mmunofl uorescence analysis of annelxn-v-f!ous Western blot data revealed that p53 and p21CIP1/Waf- l level increased strongJy upon 5'-NIO treatment , The a uthor next tested whether 5'-NIO could activate apoptos is related proteins s uch as caspase-3/-7/-9, [n cells exposed to 5‘-NIO, activation of caspase-3 and -7 was observed, Interesti ngly‘ caspase-9 and cytochrome c cou ld s li ghtly activate in response to 5’-NIO. These results indicate that 5’-NIO strongly induces oral cancer cells apoptosis via a Mi tochondria-mediated cas pase cascade pathway, These observations together s uggest that 5’ NTO may have a possible therapeutic potentia[ to oral cancer
Indirubin is the ac ti ve ingredi ent of a traditional Chinese herbal medicine, Danggui Longhui Wan, used for t he t reatment of chronic myelocytic leukemia Here, we report that novel indirubin ,- 11‘ ivative‘ 5’ - nitro-indirubinoxime (5’ NIO) , has potent anti- proliferative activity on va rious human cancel‘ cells and oncogenic RK3E- ras rat kidney cells with ha lf- inhibi tory concentrati ons (1C50) ra nging from 1- 12 M, Treatment with indirubin derivative induced the activation 01' caspase 7 rollowed by apoptos is in RK3E- ras cells. lndirubin derivative showed strong anti-tumor activity in rat solid and oral tUll10r models , Direct inj ection of indirubin deri vative every other day for 10 days induced signifi cant inhi bition of tumor growth in Sprague-Dawley rats bearing RK3E- ras-induced tumors Histologically. t reatment with indiru bin de ri vative caused s ignifi cant inhi bit ion of tumor formation with increased apoptosis and decreased tumor cell prolife ration, These f indings provide the potent ial va lue of indirubin deri vative a s a novel candidate for ant i-tumor agents
PDT is an establi shed cancer treatment modali ty , This can be attributed to the attractive basic concept of PDT; the combina ti on 0[' two ther a peut ic agents, a photosensitizing drug and light, which are r elatively harmless by themselves but combined ultimately ca use more 0 1' less selective tumor destruction, The bacteriochlorophyll - derivatived photosensitizers are known to be s ta ble and hi ghly efficient‘ In this s tudy, we conducted a series of experiments to develope the ligh t induced anticancer drugs against oral cancer cell ‘ We tested the cytotoxicity of the hydroxybact eriochlorine by MTT a ssay and observed the cell death pattern (apoptosis or necrosis) after PDT by hoechst 33342 and propidium iodide s taining methods , IC50 value of the hydroxybacteriochlorine was 31,3ngjm.Q, At higher doses of hydroxybacteriochlorine () 60 ng/ 뼈) ， cancer cells died exc lus ively by nec rosis after PDT By contrast, at IC50 value, h ydroxybacteri ochlorine in duced ca ncel' cell to undergo a poptotic cell death The induction begins approximately 6 hours after PDT We investigates int racellu la r localizati on of hydroxybact eri ochl orine by ora l cancer cell via confocal laser scanning microscopy, Oral can cer cells dual-stained with hydroxybactel' iochlorine and organelle-specific fluoresc ence probes (Mi totracker, Lysotracker , ER- Trac ker) revealed an int l'acellula l' flu orescence distribution restrict ed to cytoplasmic compartments with no detectable fl uoresce nce in the nucleus Confocal images of cells containing hydroxybacteriochlorine were never overlap to mi tochondria, lysosome, endoplasmic l'eticulum when digitally overlapped with the organelle-specific flu orescence probe images of the same cells , These resul ts demonstrat ed that the hydroxybacteriochlorine may have a function as a photosens it izer and cytotoxicity hydroxybactel' ioc hlorine for oral ca ncer cell is more sensitive than head & neck cancer cell or cervical cance l' cell Ther efore PDT using hydroxybact eriochlorine is suitable treatment for oral cavity car cinoma patients.
Chromosomal abnormality s uch as aneuploidy is one of the main factors to cause cancers This abnormality is caused by defects in centrosomal duplication‘ and most cancer cells have extra copies of centrosomes, r esulting in t he formation of multipolar spindles. Several kinases playing in mitotic phase have been implicated in regulating the centrosomal cycle‘ spindle checkpoint‘ and chromosome co ndensation and segregation. They also have Lhe ability to act as oncogenes. FOl studying the relationship between rnitotic kinase and oral cancers, the kinase activity of polo-like kinase-1, which is one of mitosis-specific kinases, is analyzed in oral carcinoma cells originated differently. Kinase activity was increased in these cancer cells compared to normal human gingival fibroblast primary cultured cells Moreover. the mitotic cell populations were a lso increased in these cell lines. Whereas the inhibition of Polo-like kinase-1 by C-terrninal domain in human gingival fibroblast cells induced multiploidy‘ the apoptotic cell population was increased in oral cancer cells overexpressed C-terminal domain 0 1' Polo- Ii ke kinase-1. These data suggested that polo-like kinase-1 might be involved in the on cogenic effect in oral cancer like other solid human carcinomas, and be target molecules for anti-cancer drug.
Nowaday many researches has proved that glutaradehyde(GA) is more excell ent medicament in vital p띠 potomy practice than formocresol (FC) . But a number of dental practitioner prefer to use formocresol in vital pulpotomy procedure todays And thus author reeva luate proper ties of gluteraldehyde and formocresol through implantation into epidermis and trypsin digestion after f ixation at 2% buffered glutar aldehyde and Burkley's formocresol solution.. And then prepared t issues were s tained by H&E a nd Masson-TI’ ichome method. GA showed definite fixative zone and γery low diffusi ble property in epi dermis and pulp t issue as glutaraldehyde and formocresol were compared. GA r epresented exceedingly lower antigenici ty than FC GA fixation exhibited more resistibility to trypsin digestion than FC. As considering these results‘ it concluded that GA would be extremely supe ri 이 . medicament to formocresol in vital pulpotomy proce
Dentin is a mineralized tissue formed by odontoblasts that are differentiated from ectomesenchymal cells , The molecul ar mech anism of odontoblast diffe rentiation remains unclear, Amino acid transporters play an important role in s up plying nutri tion to normal a nd ca ncer cells including odntoblasts, and for cell proliferation , Amino acid transport system L is a maj or nutrient t ransport system responsible for the Na+' -independent transport o[ neutral amino acids incJuding several essentiaJ amino acids , The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2) , In this study, the expression pattern and role of amino acid transport system L were, therefore, investigated in the differentiation of MDPC-23 cells derived from mouse dental papilla celJs , To determi ne the expression Jevel o[ amino acid transport system L participating in intracelJ ular transport of amino acids in the differentiat ion 0 1' MDPC-23 cells, it was examined by RT-PCR, observation of cell morphoJogy‘ A1izaline red-S staining ancl uptake analysis after inclucing experimental differentiation in MDPC-23 cells The res ults were as follows , The LAT1 mRNA was expressed in the early stage of MDPC-23 cell differentiation , The expression leveJ was gradually increased by time course and it was decreased after the late stage, The LAT2 mRNA was not observed in the earJy stage of MDPC-23 cell differentiation, The LAT2 mRNA was expressed at the 11 days 0 1' MDPC-23 cell differentiation and the expression level was gradually decreased by time course, There was no changes in the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during the differentiation of MDPC-23 cells , The expression of ON mRNA was graduaJJy decreased but the expression of ALP mRNA was increased during differentiation of MDPC-23 cells , The L-Ieucine uptake was increased by time cour se from the early stage to the 9 days in MDPC-23 cell differentiation , The amount of L-Ieucine uptake was maintained to the 11 and 14 days of MDPC-23 cell differentiation As the resul ts‘ it is considered that among neutral amino acid transport system L in differentiation of MDPC-23 cells , the LATl has a key role in cell proliferation in the early stage and middle stage of cell differentiation and the LAT2 has an important roJe in ceJJ differenti ation and mineralization in the Jate stage of cell differentiation for providing cells with neutral a mino acids incJuding several essentiaJ amino acids
Ameloblastic carcinoma is an ameloblastoma wi th histological malignant transformation with 01' without metastatic di sease_ We report an ameloblastic carcinoma ex ameloblastoma of right mandibular body in 7 -year-old girl with a heterogeneo us hi stologic components_ The tumor s howed so lid s heets composed of atypical kerati nized squamous cells. small ovoid cell s. and s pind le cells in addi tion to a bit of beni gn ameloblastoma component_ Immunohis tochemically, the squ amous cells were strongly cytokemtin posi ti ve/vimentin negati ve and the small ovoid and spindle ce lls were weakJy cytokeratin positi ve/v imentin pos it ive_
The incidence of oral cancer varies widely worldwide. Its treatment often produces significant functiona l and cosmetic morbidity and its mortality continues to be high. Strategies for diagnosing, managing and preventing the disease must be based on an understanding of r isk factors and the pathogenesis of the disease locally. The World Health Organizat ion has documented that few countries have a comprehensive a pproach to the diagnosis , management and prevention of cancer at various anatomic sites. This presentation will review the United States’ experience with oral cancer and its strategies fo1' affecting incidence and prognosis . The greatest obs tacles are often political and economic, not scientific.
Oral squamous cel1 carcinoma(OSCC) is the most common malignancy of head neck region. Typically OSCC cells s how persist ent invasion that frequently leads to local recurrence and distant lymphatic metastasls However, molecular mechanisms of invasion of OSCC remain poorly understood. Her e we identifi ed periostin, interferon induced transmembrane protein l (IF1TM1) and wingless-type MMTV integration site family, member 5B(WNT5B) , as invasion promoting molecules in OSCC by comparing gene expression profiles between a parent OSCC cell line(MSCC-l) and its highly invasive clone(MSCC-1nvl). Overexpression of periostin, IFITMl and WNT5B mRNAs were confirmed in MSCC-1nvl by RT-PCR. Transfection of these molecules promoted invasion of OSCC cells Moreover , siRNA t r eatment of these molecules suppressed invasion of cancer cells in vitro I nter estingly, Periostin, 1F1TMl and WNT5B were highly expressed in OSCCs in comparison with nonnal tissues. 1n an orthotopic mouse model of OSCC, periostin-overexpressing cells metas tasized spontaneously to cervical lymph nodes and t o t he lung through their aggressive invasiveness. These findings suggest that peri ostin, IFITMl and WNT5B play important roles for invasion and of OSCC and can be prognostic markers and therapeutic t argets of OSCC.
Oral mucosa is made of stra tifi ed oral epithelium(that forms the surface) and subjacent connective tissues Alt hough oral mucosa lines the oral cavity, and thus provides a barrier between external and inte1'na l(tissue) envi1'onment, it exp1'esses regional va1'ia tions in its clinical appearance based on its structu1'e(ker atini zation/ non- ke1'atini zation; nature of connective tissues ; vascul arity, etc.) . The structu1'e of or al mucosa a lso reflects its flln ctional adapta ti on. Available liter ature indicates t hat oral mucosa may be viewed as masticat ory, lining 0 1' specialized types, aJl of which remains continuously moist and unexposed to light. Oral mucosa, however, is subj ected to daily wear and tear due t o the natllre of food , environmental chernicals , t oxins and met abolites. Hence, an understanding of the biological behavior of its constituents cells and ext1'acellular matrice s , tha t• are in constant s tate of adapt ive changes, is usefll l. Oral epithelium is a multi - Iayered and multi - functional dynarnic system that separates the o1'al environment f1'om the va1'ied nature of the s llbj acent connective tlsslles The connective tissue is eithe1' loose or dense collagen ous, muscula r or osseous, and may contain glands. Thus,oral epithelia play a critical 1'ole in the mainten ance of homeost atic effi ciency of the environment of both the o1'al cavi ty and subj acent connective tisslles. The structure and fllncti ons of the oral epithelial cells are regulated by a complex interplay between genetic and environmental fa cto1's, the knowl edge of which is unfolding only recently. The fundamental prernise on whi ch 3-dimensional organization of oral epithelial integrity and st a bility are maintained is the principl e of st eady-state syst em. The basic featU1'e of the steady- st ate syst em is t he 1'egulation of cellula1' and t issue homeost atic mechanisms. Since during its ontogeny, oral epithelial cells are subj ect ed to daily wear and tear , resulting from external mechanical and chernical facto1's, a 1'eservoi1' of cells, with regene1'ative capacity(va1'iously labeled in lite1'ature as p1'ecurso1' ceJls, p1'ogeni tor cells OI‘ stem cell s) that can maintain the steady-state system of oral epi thelia, are 1'equired. Such a reservoir of cells are located in the basal layer of oral epithelia. These cells proliferates, differentiate into ker atinocyt es, and are lost through desquamation or a poptosis to maintain the steady-state syst em and thus equilibrium of tissue homeostasis
Down-regulation of E-cadherin marks the initiation of the epithelial-mesenchymal transit ion (EMT), a process exploited by cancer cells that display an invasive phenotype. The zinc finger transcription factor , Snail , functions as a potent repressor of E-cadherin expression that can, acting alone or in concert with the Wnt/β -catenin/TCFaxis, induce EMT both in neoplastic cells as well as normal cells during embryonic development. While mechanisms that might coordinate signaling events initiated by Snail and Wnt remain undefined, it is demonstrated that Snail displays ß -catenin-like canonical motifs that support its GS3Kdepende nt phosphorylation, ß - TrCP-directed ubiquitination and proteasomal degradation. Accordingly, Snail half- life and repressor activity is enhanced by constructingphosphorylation- res istant mutants, depressing GSK3 activity via the Wnt signaling cascade or inhibiting proteasome function These findings define a potential mechanism wher eby Wnt signaling stabilizes the intracellular Snail and ß -catenin in tandem so as to cooperati vely engage the transcriptional programs that control EMT 1n human cancer, hyperactive Wnt signaling leads to the formation of a bipartite, -catenin/T ce ll facto r (TCF) complex which has been postulated to serγe as a trigger of the EMT that characterizes the tissue-invasive and metastatic properties of cancer cells. However, the molecular mechani sms by whi ch the ß -catenin/TCF complex triggers EMT-like programs remain undefined. Herein, it will be demonstrate that canonical Wnt signaling engages tumor cell de-differentiation and tissue-invasive activity by a heretofore unsuspected Axin2-dependent pathway that serves to stabilize the zinc transcription factor, Snail - a key regulator of normal and neoplastic EMT programs. Axin2 exerts this effect by acting as a nucleocytoplasmic chaperone for GSK3, the dominant kinase responsible for controlling Snail phosphorylation, ubiquitination and proteosomal degradation. As dysregulated Wnt signaling marks a diverse array of carcinogenic and metastatic states, t he identification of a ß -catenin/TCF-regulated Axin2/GSK3/Snail1 axis provides new mechanistic insights into Wnt- mediated EMT programs of human cancer
Al t hough substance P(SP) , a potent pro- inflammatory peptide, is involved in inflammation and immune responses‘ t he eff'ect of SP on t he expression of macrophage inflammatory protein 3a (MIP- 3α CCL20) in periodontal liga ment(PDL) cell s a re unknown, Equally as enigmatic is the link between SP, t he stress protein heme oxygenase- l(HO-l) ‘ and CCL20 procluction, We investigated whether SP induces the release of chemokine CCL20 from immortal ized PDL(IPDL) ceJJ s‘ and fur ther c l a꺼 SP mediated pathways, We also examined the relationship between HO-l a ncl CCL20 by t reating PDL cells with SP, Incubating IPDL cells with SP increased expression of CCL20 mRNA a nd CCL20 protein in a dose-time dependent manner Highly selective p38 and ERKl/2 inhibitors abrogated SP-induced expression of CCL20 in IPDL cell s, SP is a lso responsible for ini t iating phosphorylation of I/C B, degradation of Iκ B‘ ancl activat ion of NF'-/C B, SP induced expression of HO-l in both a concentration- and time-dependent man nel ‘ and CCL20 refl ected s imilar patterns, The inductive effects o[ SP on HO- l and CCL20 wer e enhanced by HO- j inducer hemin and the membrane-permeable cGMP analog 8-bromo-cGMP, Conversely, this pathway was inJübited by t he 1-10난 inhi bitor zinc protoporphyrin IX(ZnPP IX) and the selective inl뼈itor of guanylate cyc1ase‘ lH-[l , 2, 4Joxad iazole[4‘ 3-aJquinoxal in-l-one (ODQ) , We report herein the pathway that connects SP along with other modulators 。f neuroimmunoregulationto the induction of HO-l and t he inflammatory mediator MIP-3a /CCL20 in IPDL cell s‘ which play an important role in the development 01' periodontitis or inflamrnation during orthodontic tooth movem
Epithelial myoepithelial carcinoma(EMC) is an uncommon malignant sali va ry gland neoplasm, compri sing about 1% 。f salivar y gland neoplasms, They are histologically composed 01' biphasic cell s such as myoepithelial cells and ductal cells EMC occurs predominantly in the major salivary glands, par t icularly in the par otid gland, They have been reported to occur onJy 10-15% in t he intraoral minor salivary glands, The authors experi enced 4 cases of EMC in Department of Oral Pathology in Seoul National University Dental Hospital from 1995 to 2007‘ and reported them with revi ew of li terature, They occurred at the age [rom 34 to 75 years with average age of 54, Three cases occurred in f'emale and 1 in male, showing predominant occurrence in female, Al I of them occurred in the f100r of mouth Three patients presented localized swollen mass at the time of admission, One patient manifested pain with surface n ecrotic ulcer, and others did not complain any symptoms, The duration of symptoms before diagnosis has ranged f rom three mont hs to 2 years in our cases Microscopically, they growed in double layered ductal structure composed 。f' ductal cells of the inner luminal layer which showed positive immunohistochemical reaction to cytokeratin and rnyoepithelial cells 01' the outer peripheral layer identifi ed by the positive reactivity to S-100 and srnooth muscle ac tin antibody, They did not show perinem al invasion‘ but invasíve growth into adjacent tissue, AII of them did not show in vasion into t he underlying bOl1e While 3 pat ients were treated with total excision of tω110 1' mass wi thout n。 evidence of recurrence a ncl metastasis, a 75 year old patient gave up receiving t reatment at the t irne of diagnos is, and then died 01' the cancer 5 years after init ial diagnosis, It seems that EMC of the intraoral minor salivary gland is a tumor 01' low grade malignancy with low potent ial of recurrence and metastasis
The human ELAV(embryonic lethal abnormal vision)-like protein HuR stabilizes a certain group of cellular mRNAs that contain AU- ri ch elements in theil‘ 3’• untranslated region, Dysregulation of mRNA s tability may be relevant in tumor biology and may lead to abnormal expression of several proteins in malignant tumors, The aim of this study is to identify the differentially expressed proteins according to the functi onal activi ty of HuR Methods : We used stabl e expression of small interfering RNA(siRNA) of HuR gene to inhibit the expression of HuR in human ora l car cinoma cell lines‘ KB cell line and YD10B cell line‘ and compared the proteomic changes between s iRNA-treated and cont rol cell line using two-dimensional gel electrophoresis , Flow cytometry caliber scan(FACS) was employed to investigate the effects of HuR on cell apoptosis and proliferation , Results: Seventeen differentially expressed proteins between the two cell lines were identified by electrospray ioruzati on quadrupole time-of- fl ight mass spectrometry(ESI-Q-TOF MS) and database searching, Among them there are eleven proteins which are significant ly up- regulated in siRNA- treated cell line, which include heat shock protein 10(influencing nucl eocytoplas rnic transpo 다， cell dedifferentiation , and inhibition of apoptosis) , keratin 19(basal cell differ ent iat ion) ‘ and nucleoside diphosphate kinase B(G protein activator), etc Enolase 1 and Ml of pyruvate kinase a re the representatives of signifi cantly down-regulated proteins in siRNA-treated cell 11l1e Conclusion : Our data suggest that HuR participate in mRNAs stability of proteins that have the counter effects in the carcinogenesis of oral ca ncer , And the functiona l proteomics a re needed to elucidate the detailed interactions between HuR and t hese molec ules
Wnt genes encode a large fami ly of secreted cysteine-rich signaling mol ecules involved in cell growth‘ differentiation and tumorigenesis, Wnt5a, a non-transforming member of the Wnt fa mily behaves as a putative onco 一 gene in many cancers including melanomas, The aim of our study was to determine Wnt5a ex pression pattern in pnmalγ oral mucosal melanomas(Ol\αÆ) and correlate it with tumor thlckness Archival ti ssues from 18 OMM cases were subjected to immunohistochemical detection of Wnt5a by the streptavidin-biotin method , These were categorized into tumors of (4 mm(thin and intermediate thickness lesions) and )4 mm(t hi ck lesions) t hickness, Most OMM cases(17/18; 94, 4%) stained positive for Wnt5a, though heterogeneously, Seven t hi ck(7/11: 64%) and one in termediate thickness(1/7‘ 14%) ol\α‘ demonstrated strongly positive Wnt5a staining(P(O, 05), The only Wnt5a-neg ative case was a thi ck OMM without local recurrence after treatment Strong Wnt5a express ion at tumor adva ncing sites suggests a role in local tumor spread, Identification of pleomorphi c epitheli oid and spindle cells as mel anoma cell populations with the most pronounced Wnt5a staining suggests that Wnt5a overexpression inJ] uences cellular phenotype, These results taken together suggest that Wnt5a is up- regulated in OMM a nd may play a role in tumor progression, Wnt5a activity is most frequently detected in advanced OMM suggesting its function in tumor progression, Expression level of Wnt5a in OMM correlated with tumor grade and t hickness
Although a number of molecules have been implicated in the metastasis of oral squamous cell carcinoma (OSCC) , the precise molecular mechanisms that deterrnine the direction of rnigration and invasiveness of OSCC cells into the lymph nodes remain unclear, Chemokines are a superfarnily of small structurally related heparin- binding proteins‘ which have been identified as attractants that control the rnigration of leukocytes‘ especia lly during imrnune and inflammatory reactlOns Moreover, recent studies have demonstrated that several types 。f cancer express chemokine receptor‘s and use chemokines to metastasize to the target organ, However, there h ave been few reports on biological behaviors by downregulation 0 1' CXCR-4 in ora l cancel‘ cells We tried to screen several OSCC cell lines in order to obtain a suitable cell line model which had the cha ract eris tic of the constitutive ly expressed state of CXCR• 4 Of the several OSCC cell lines, only KOSCC-25B showed the high expression of CXCR-4 in both RT-PCR and Western blot analysis, siRNA-CXCR-4 infected subclones of KOSCC-25B(Si, 3‘ Si 1이 showed downregulation of CXCR-4 expression as expected‘ At serum-free co ndi tion‘ Si.3 s ubclone s ignif icantly decreased cell proliferations at 24 h and 48h and Si, lO subclone significant ly dec reased cell proliferations at 24 h Si ,3 clone dec reased to 67 ,4% and Si,lO clone to 65 ,5% in comparison to vector infected cells These data suggest that the downregulation of CXCR-4 expression could induce anti-rnigratory and ant i- rni g ratory effect
Apoptos is s ignal- regulating kinase 1 (ASKl) is a rnitogen-activated protein kinase kinase kinase(MAP3K) 떼 th proapoptotic functlOn 1'he kinase activity of ASKl is stimulated by a variety of death signals , including 1'NFα • Fas ligation, reactive oxygen species, and antineoplastic agents, ASKl promotes cell death by activating the c- Jun N-termina l kinase/stress-activated protein kinase MKK4/MKK7-JNK/SAPK pathway and MKK3/ 1\αCK6-p38 pathway‘ ASKl activity is highly controlled in cells by multiple mechanisms, including phosphorylation, oligome ri zation, and protein- protein ll1 teractlOns Epigallocatechin-3-galla te(EGCG) is the major bioactive polyphenol present in green tea, It possesses anti-oxida nt , a nti - mutagenic‘ a n ti - prote이 ytic ， and anti-proliferative activity, In addition. it has been shown to inhibit cyclin activity, and inhibit cell cycle progression 1n the present study, we exarnined the effect of EGCG on ASKl- overexpressed cells , We expected that EGCG contributes to cell a poptos is by activating ASKl functlOn However, EGCG showed no suppressive effect on cell s urvival of ASKl-overexpressed cells and seemed to promote cell survlval Importantly, the EGCG treatment in creased Akt activity when cells expressed enough amount of ASKl protein, These results s uggest that the presence of ASKl may modify the inhibitory effect of EGCG on cell survival through Akt pathway,
[n order to obtain the trlle expected DNA prod uct from PCR and RT-PCR using genornic DNA or cDNA reversely transcribed from mRNA. the PCR should be done in an appropriated condition. Sometimes the PCR was repeatedly fail ed. and cventllally the PCR product was turned out to be nonspecific and rudimentary . And more‘ t he PCR prodllctwas not reproducible even though careflll repeat of experiments. As the PCR was based on the exact primel hybridization. the condition of primer hybridization should be properly controlled by a nnealing temperatllre. But the selection of primer seqllences for targeting a specific gene is mostly important. A new method of primer eval uation is now available llsing DNA base pair polarity program. This study presents an example of PCR targeting to human Bax gene using genomic DNA. The DNA base pair polarity theory can di vide the genetic cord into propel DNA segments and calclllaLe their DNA base pair hybridization energy. Thus. mathematically the degree 0(' exact primer hybridization can be expected for the t r1l8 targeting of PCR. However, the DNA base pair polal'ityanalysis demonstrates that the more frequent number of DNA segment incl'eased the specificity of PCR. but decreased its sensitivity . While the greater polarity of DNA segment composed of increased nllmber of polarized DNA base pairs showed increased sensitivi ty 0 1' PCR. bllt relati vely decreased specificity of PCR. With the mllltiple analysis of PCR. especially for PCR cloning from the gDNA and cDNA, we found that the primers themselves showed secondary strllcture of partial hybridization between sameprimers or each pair primers. The DNA base pail‘ polarity signal can directly demonstrated symmetric sequences 0 1' each primer. and also can distinguish the dimmer formation from each pair primers. At least the symmetric seqllence of fOlll‘ base pairs dramatically showed the dimrner formation. On the other hand. in addi tion Lo the statlls of DNA base pair polarity the three-dimensional strllctllre of DNA dOllble helix targeted by the primer seqllences may affect the sensitivity and specificity of PCR detection. The present study introduced a new method of primer evalllation and selection in order to obtain abundant and exacL! y-trlle DNA product for genomic ffilltation analysis and gene expression profï le
Al though the changes in tooth morphology and hardness by hydrogen peroxide(H20 z) have been r‘epor‘.ted .‘ the pαr。o야t뻐ec야tive role of heme oxygenase-l(HO-l) against the cytotoxic effects of H202 has not been clarifïed i n human pulp cells ln this st udy. we investigated whether HO-l is involved in Hz0 2-induced cytoLox icity a nd examined the production 0 1' dentin sia lophosphoprotein(DSPP) and other mineralization markers‘ in hllman pu lp cells H202 decreased cell viabi lity, but increased HO-l and DSPP expression in a concentra tion and time dependent manner . Inhibitors of gllanylate cyclase. PI3K, ERK. and p38 MAP kinase blocked H202-indllced cytotoxicity and the expression of HO-l and DSPP mRNAs in pulp cells. These data suggest that the induction of HO-l by H202 in plllp cells plays a protective role against the cytotoxic effects 0 1' HzOz and stimulates DSPP expression‘ reslllting in prematllre odontoblast dilTerentiation throllgh pathways that involve cGMP‘ p38. and ERK.
The aims 0 1' t his study we re to evaluate proliferative and apoptotic cell distribution pattern in condylar articu lar and pl'oliferative layer as two types of diet(soft and ha rd) we1'e supplied to growing 1'a ts, 30 male Sprague Dawley g1'owing l'ats(twenty-one c1ays fl'om bi l'th) were c1ivided into two group, Each 15 rats were supplied by soft and hal'd di et , After c1 iluted Bl'du solution(5mg/ Kg)was injected into pel'itoneum befol'c sacrificc. rats wel'e sacl'i f iced at 1. 2. 3 week llltel'vals St1'eptoavidin-Biotin method for Brdu was used to evaluate celluJar proliferative activity. ancl fluol'escent TUNEL method was used to estima te the apoptotic activity, The results about this expe riment wel'e recorded about ante1'io1' and posterior condyle separateJy, In anteriol' portion of condyle, soft diet group showed inc1'eased p1'olife1'ative celluJar activity tha n hal'd diet group dw'ing 1, 2 week but decl'eased than hal'd diet g1'oup at 3 weeks. while hal'd diet group showed constant p1'oliferative cellular activity during all experimenta l period , ln posterior pOl'tion of condyl e、soft diet group showed increased activity than hard diet group at 3 weeks, while ha rd di et gl'oup showed constant proliferative cellular activity during exper‘ imental period , In a nterior pOl'tion of condyle‘ soft c1 iet group showed increased apoptotic activity with time progress, but hard diet group showed continuous low level of activity during experimental period , In posterio1' porti on 0 1' condyle, hard diet group showed cons tant low level apoptotic activity, plthough showed the lowes t leveJ at 1 week, On the con trary soft diet g1'oup showed c1ecl'eased apoptotic activity wi th time progress, ConcJusively, in soft diet group an teroposterior direction was reduced in condyJar morphologic dimension because of decreased prol iferative cellular and increased apoptotic activity on anterior portion and vertical dimension 0 1' condyle was increased because of increased proliferative cellular and decreased apoptotic activity on posterior portion. but in hard di et group proli ferative cellular and apoptotic activity were comparatively constant. and thus harcl diet group s howed antet'opos teriorJy broad and flat condylar morphoJogy