Human saliva contains a large number of proteins and peptides whose composition may alter as a consequence 。f disease. To date. however‘ the proteins and peptides that routinely populate thi s ora l fluid a re largely unknown To provide a catalogue of saliva proteins, we have surveyed the unstimulated human whole saliva by us ing shotgun proteornics. For the shotgun approach, whole saliva proteins were digested into peptides with ChemDigestD ‘ and the resul ting peptide fragments were separated by RP-HPLC, followed by each fraction was t ryptic digestion. ChemDigestD-Trypsin digested peptides were analyzed by tandem mass spectrometry(MS/MS) us ing a nano-LC eq 버 pped quadrupole-time of f1ight mass spectrometer, and the obtained spectra were searched against human protein seq uence data base using MASCOT. Shotgun proteomics allowed a total of 291 human pr。 teins to be confidently assigned . The largest group(17 .2%) of the identified proteins sorted into functional catego ries was included in t he signal t ransduction funct ion except for the hypothetical 0 1' unknown function. This work provides a valuable s ta rting point for the analysis of human salivary proteins and their biological functions and candidates from human whole sali va that may prove to be of diagnostic and therapeutic significance

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

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_

Human saliva conta ins a la rge number of proteins and peptides whose composition may alter as a conseq uence of disease. '1'0 date‘ however. the proteins and peptides that routinely populate t his oral fluid are largely unknown, '1'0 provid e a ca ta logue 이， sali va protei ns. we have surveyed the unstimulated human whole saliva by using shotgun proteomics. F'or the shotgun a pproach‘ whole sali va proteins were digested into peptides with ChemDigestD and the res ulting pe ptide fragments were sepa rated by RP- HPLC, followed by each fraction was tryptic di gestion ChemDiges tD- Trypsin diges ted pe pt ides were analyzed by tandem mass spectrometry (MS!MS) using a nano-LC equi pped quadru po le-time of fli ght rnass spect rometer, and the obtained spectra were searched against human protein sequence da tabase us ing MASCOT Shotgun proteomics a llowed a total of 291 human proteins to be confidently assigned. The largest gro u p (17 , 2%) of the identifi ed proteins sorted into functional categories was included in the s ignal transducti on function except for the hypothet ical or unknown functio n, This work provides a valuable starting point for the ana lys is of human sa l i va ry protei ns a nd theil‘ biological functions and candidates from human whole saliva that may prove to be of diagn ost ic and t herapeutic s ignif‘ Ica nce

The tumor suppressor gene, phosphate and tensin homologue(PTEN) has been shown to dephosphorylate the phosphatidylinositol 3-kinase(PI 3-K)-generated phosphatidylinositol(3-5)-triphosphate in vivo, thus interfering with the potentially oncogenic signals emanating from PI 3-K. Promoter hypermethylation of CpG islands has recently been shown to be an epigenetic change resulting in loss of function in some genes involved in cell cycle regulation and DNA repair. Immunohistochemal staining for monoclonal antibody 6H2.1 was performed from paraffin embedded blocks of 20 benign epithelial lesions and 40 head and neck squamous cell carcinomas(HNSCCs). Immunoreactivity was graded semiquantitatively by considering the percentage and intensity of the staining of the tumor cells. Also, this study tried to identify PTEN methylation in benign epithelial lesions(24 cases) and HNSCCs(44 cases of paraffin embedded blocks, 4 cases of frozen tissues) using methylation-specific PCR(MSP). In HNSCCs, immunoreactive scores of stage 1 and 2(12 cases, average score 85.2) were higher than those of stage 3 and 4(15 cases, 41.9) and statistically significant(P=0.017). Immunoreactive scores of moderate and poorly differentiated carcinomas(22 cases, 61.6) are more or less lower than those of well differentiated carcinoma(15 cases, 87.0) but not significant(P=0.361). Among 24 cases of benign epithelial lesions, 12 cases showed unmethylated PTEN but none methylated. In HNSCCs, 22 of 44 paraffin embedded blocks showed unmethylated PTEN but none methylated, and all 4 frozen tissue revealed unmethylated PTEN, one of which(25%) methylated. We consider that the loss of PTEN protein expression may be associated with the progression of HNSCCs and the other alteration rather than methylation may be important in the inactivation of PTEN in HNSCCs.

Genomic imprinting is defined as parent-of-origin expression of specific genes and may play an important role in embryonal development of mammals. Loss of imprinting(LOI), biallelic expression of the imprinted genes, have been observed in a variety of human tumors and syndromes. H19, a paternally imprinted gene, is transcribed as an untranslated RNA that serves as a riboregulator. LOI of H19 is observed in a variety of human malignancies. In this study, LOI of H19 was examined in head and neck squamous cell carcinomas(HNSCCs). Four(28.6%) of the 14 HNSCCs and 8(28.6%) of the 28 inflammatory oral lesions were informative for imprinting analysis of H19. H19 was imprinted in all inflammatory oral lesions, however, 2(50%) of the 4 informative HNSCCs manifested LOI. These data suggest that LOI of the H19 may play a role in the oncogenesis of HNSCC.

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,

Metastatic spread to cervical lymph nodes(LNs) is a major determinant of outcome in oral squamous cell carcinoma (SCC). To provide an useful method for the detection of lymph node micrometastases, we fulfilled the histopathological examination and reverse transcriptase polymerase chain reaction(RT-PCR) using the paraffin-embedded LNs of oral SCC patients. In this study, 78 LNs from 12 patients with primary oral SCC were analyzed. Metastases in the regional LNs were evaluated by RT-PCR for squamous cell carcinoma antigen(SCCA) and cytokeratin 5(CK5). Detectability of metastatic LNs by RT-PCR was compared with histopathological examination. Of 78 LNs, CK5 and SCCA mRNA were detected in 32(41.0%) and 8(10.3%), respectively. Histopathologically, 10(12.8%) of 78 LNs were positive. CK5 mRNA was detected in all 10 histopathologically positive LNs. In contrast, SCCA mRNA was detected in 5 of 10 histopathologically positive LNs. These findings suggest that genetic diagnosis by RT-PCR based on CK5 mRNA expression may be sensitive and clinically useful technique to detect the presence of metastatic carcinoma cells in regional lymph nodes of oral SCC.