Urokinase-type plasminogen activator(uPA) bound to urokinase plasminogen activator receptor(uPAR) expression is strongly correlated with the metastatic potential of various tumors by enhancing ECM degradation through plasminogen and matrix metallopreotease activation. But expression of uPA/uPAR in human malignant salivary gland tumors has been rarely reported. The purpose of this study were to investigate mRNA expression and cytologic concentration of uPAR in SGT cell line compared to various cancer cell lines by RT-PCR and ELISA method, and to study migration and adhesion assay. These results would be to apply the pathogenesis and prognosis of malignant salivary gland tumors. All the cell lines(SGT, HN 4, SCC 25, and HeLA) were cultured under DMEM with 10% FBS at 37℃ in a 5% CO2 incubator. We studied a possible association between mRNA expression and cytosolic concentrations of uPAR in SGT cell line compared to various cancer cell lines using RT-PCR and an enzyme-linked immunoassay( ELISA) method. And also cell adhesion and migration assay were done in all the cell lines. In migration assay SGT cell line was about 2.5-4 folds higher than another cell lines. In adhesion assay SGT cell line was about 1.1-2 folds higher than another cell lines. uPAR cytosolic concentrations of SGT cell line was about 3.4-10 folds by ELISA, while mRNA expression was about 2.5-5 folds by RT-PCR. Oral Scc cell lines showed the lowest value. uPAR protein and mRNA expression were correlated with migration and adhesion assay. From the aboving results, high cytosolic concentrations and mRNA expression of uPAR were correlated with migration and adhesion assay. It suggested that this might be a specific marker for malignant potential of SGT cell line and would be contributed to pathogenesis and prognosis of human salivary gland adenocarcinoma

Plasminogen activator(PA) system such as urokinase plasminogen activator(uPA), urokinase PA Receptor(uPAR), tissue, tissue PA, and PA inhibitor-1&2(PAI-1&2) play a role in tumor invasion, metastasis, and proliferation. It is interested that these factors in patients with primary oral squamous cell carcinoma(Oral SCC) will be evaluated and correlated with clinicopathologic variables. Recently, these expression of primary oral SCC has been restricted to clinical or immunohistochemical study such in vivo study. The purpose of this study were to investigate the mRNA expression and cytologic concentration of uPA, uPAr, tPA, and PAI-1,2 in oral SCC cell lines compared to NHOK and to apply these results to evaluate early detection biomarkers of oral SCC in future. All the cell lines(NHOK, HN 4 and SCC 25) were cultured under KBM bullet kit at 37℃ in a 5% CO2 incubator. We studied a possible association between mRNA expression and cytosolic concentrations of uPA, uPAR, tPA, and PAI-1,2 in oral SCC cell lines compared to NHOK using RT-PCR and an enzyme-linked immunoassay(ELISA) method. uPA mRNA expression was about 5-6 folds, while uPAR was a bout 3 f olds, and PAI-1 was about 1 .5-1.6 f olds. PAI-2 was a bout1.2 -1.3 f olds t han that o f NHOK, w hile t PA w as l ower t han that of NHOK. uPA cytosolic concentrations was about 15-19 folds, while uPAR was about 8 folds, and PAI-1 was about 3-4.5 folds. PAI-2 was about 2 folds than that of NHOK, while tPA was lower than that of NHOK. Both uPA, uPAR, and PAI-1,2 cytologic concentrations were correlated with mRNA expression of oral SCC cell lines. From the aboving results, high cytosolic concentrations of uPA, uPAR, and PAI-1 & 2 were correlated with mRNA expression. It suggested that these might be specific markers for oral SCC cell lines and these results would be contributed to evaluate early detection biomarkers for human oral squamous cell carcinoma.

Tumor cell biological factors, such as urokinase plasminogen activator(uPA) and its inhibitor plasminogen activator inhibitor- 1(PAI-1) play a role in tumor invasion, metastasis, and proliferation. These factors in patients with primary oral squamous cell carcinoma(Oral SCC) will be evaluated and correlated with clinicopathologic variables. However, relatively rarely has been known in oral squamous cell carcinoma in vivo and in vitro study . The purpose of this study were to investigate the protein expression of uPA and PAI-1 in oral SCC cell lines cell line compared to NHOK and to study migration and adhesion assay. All the cell lines were cultured under KBM bullet kit at 37℃ in a 5% CO2 incubator. We studied a possible association between cytosolic uPA and PA-1 concentrations in oral SCC cell line compared to NHOK using an enzyme-linked immunoassay(ELISA). Cell adhesion and migration assay were done in all the cell l ines. In migration assay oral SCC cell lines were about 70 folds higher than NHOK. In adhesion assay oral SCC cell line were about 7-12 folds higher than NHOK. uPA cy tosolic concentrations was about 15-19 folds and PAI-1 was 3 to 4.5 folds than that of NHOK. Both uPA and PAI-1 concentrations were correlated with migration and adhesion assay. High cytosolic concentrations o f uPA and PAI-1 were correlated with migration and adhesion assay . It suggested that these markers might be specific for oral SCC cell line and these results would be contributed to treatment and prognosis of human oral squamous cell carcinoma.

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