Autophagy is recently receiving the spotlight as the development strategy for promising anticancer drugs. In particular, the majority of anticancer drugs originating from natural products are known to induce autophagy. Saururus chinensis has been used for treating various inflammatory diseases. Recent research has revealed that the extract of Saururus chinensis possess cytotoxicity for various types of human cancer cells. However, the exact action mechanism of Saururus chinensis extract for oral squamous cell carcinoma (OSCC) has not been studied yet. Therefore, the authors of this research aim to study the effect of methanol extract of S. chinensis (MESC) on OSCC cells. To observe the cell proliferation inhibitory effect of MESC on HSC3 cells, the authors conducted the trypan blue exclusion assay. Also, the action mechanism of MESC was studied by conducting the cell cycle analysis, acidic vesicular organelle (AVO) staining and flow cytometry analysis, monodansylcadaverine (MDC) staining, propidium iodide staining, and Western blotting on MESC-treated HSC3 cells. When HSC3 cells were treated in MESC, the cell proliferation was suppressed in time-dependent and dose-dependent manners. Also, the number of sub-G1 arrested cells increased in a dose-dependent manner. MDC punctate and AVO puncta significantly increased respectively. Western blot analysis demonstrated the expression of autophagy-related proteins increased, but apoptotic proteins were not observed. Also, the pAkt protein was reduced, while the p-p38 protein and pERK protein increased. According to our results, MESC induced autophagy and accompanied changes in the cell cycle in HSC3 cells. Also, the alteration in Akt, ERK, and p38 pathways were confirmed. This result suggested the possibility of MESC as the new promising adjuvant for treating OSCC patients.

Hippo signaling is one of the signal transduction pathways revealed in Drosophila and mammalian. This signaling is known to control proliferation and growth of normal cells or cancer cells, in which many signaling proteins form a network. In this network, tumor suppressor kinases include MST and LATS while YAP and TAZ exist as oncoproteins. Combined with transcription factor, YAP the oncoprotien starts to secrete growth factors such as CTGF, FGF, and Cry61 regulating the growth of cells or the organ sizes. The YAP is also associated with the development of early embryo and the regeneration of the skin wound as well as abnormal growth of cancers in case of over-expression. Although many previous studies have found various tumors with YAP over-expressed, the expression of YAP is not yet clearly identified in oral cancer. The aim of this study was to check the expression level of YAP in oral squamous cell carcinoma. So we performed PCR and Western blot to check YAP expression in mRNA and protein level respectively. In result, all of the 13 cell lines examined has presented the expression of YAP, and especially in HSC2 and KOSCC11 cell lines has been observed the remarkable level of expression. In conclusion, we confirmed the overexpression of YAP cell line in oral squamous cell carcinoma, it will be a great help to the study carried out in the future. Once you understand the mechanism of oncoprotien YAP in oral cancer cells, it seems possible to research and development of targeted tumor therapy agents in oral cancer.

New therapeutic measure are needed to improve the outcome for patients with oral squamous cell carcinoma(OSCC) because OSCC continues to portend a relatively unfavorable prognosis. Recently RNA interference(RNAi) has emerged as an effective method to target specific genes for silencing. Although overexpression of urokinase-type plasminogen activator receptor(uPAR) has been implicated in progression and metastasis of OSCC, the transfection effect of RNAi- uPAR on OSCC has been rarely reported. The purpose of this study were to examine the efficient and specific inhibition of uPAR mRNA and protein expression by siRNA targeting of uPAR through RT-PCR and immunoslot blotting, and to study cell proliferation activity, adhesion, invasion and migration in vitro compared to the controls. In MTT assay, siRNA-uPAR transfected cells showed about 70-80% cell proliferation compared to OSCC cell lines after 2 days. In adhesion assay, siRNA-uPAR transfected cells showed about 20-30% adhesion activity compared to OSCC cell lines, but similar features to those of BSA coated wells. In migration assay, siRNA-uPAR transfected cells showed about 60% migration activity compared to OSCC cell lines, but higher 3.5 folds to those of BSA coated wells. In invasion assay, siRNA-uPAR transfected cells showed about 55% invasive activity compared to parental cell lines. mRNA expression of siRNA-uPAR transfected cells showed about 10-15 % compared to parental cell lines by RT-PCR. Protein expression of siRNA-uPAR transfected cells showed about 25% compared to parental cell lines by ELISA assay. It suggested that RNAi-uPAR tranfection might be used as a potent and specific therapeutic tool for the treatment of oral squamous cell carcinoma, especially in inhibiting invasion and metastasis.

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.

Oral squamous cell carcinoma (SCC) is one of the leading causes of cancer mortality worldwide. It is generally thought that adjuvant chemotherapy provides modest prolongation of survival in various carcinoma. Docetaxel (Taxotere, TXT) play a significant role in the treatment of various solid tumors of epithelial origin. CsA (immunosuppressive drug) was widely used as adjunct for the treatment of cancer. Thus, it is important to pursue the apoptosis of IHOK and oral SCC induced by TXT combined with CsA related to the pathogenesis of oral SCC. But TXT combined CsA effect on IHOK and oral SCC remains unclear. After cultured IHOK and HN 22 oral squamous cell carcinoma cell line treated by 10 nM TXT and 1 μM, and caspase inhobitor, respectively, apoptosis index, cytochrome c and caspase-3 -8, -9 mRNA expression by RT-PCR, and procaspase-3 protein amount by immunoslot blotting was prepared. The purpose of this study were to examine the TXT-induced apoptosis pathway via caspase activation by CsA enhancement, and to apply these results to an effective therapeutic treatment plan for oral SCC by TXT combined CsA . 10 nM TXT showed about 60%, 55% celluar apoptosis of IHOK and HN 22, cell line, respectively, while CsA alone did not induce apoptosis in IHOK and HN 22 cell line. 1 μM CsA combined with 10 nM TXT increased apoptosis in IHOK and HN 22 cell line through caspase-3 and cytochrome c mRNA expression, while could not effect on caspase-8 and -9. Caspase inhibitor suppressed apoptosis of IHOK and HN 22 cell line induced by a combination of 1 μM CsA and 10 nM TXT. Immnoslot blotting showed procaspase-3 activation by a combination 1 μM CsA and 10 nM TXT, while caspase inhibitor inhibited activation. It suggested that a combination of CsA and TXT might induce increased apoptosis of IHOK and HN 22 oral squamous cell carcinoma cell line through caspase-3 activation. This treatment with a combination of TXT and CsA may be an effective therapeutic strategy for oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is the most common malignancy and is a major cause of worldwide cancer mortality. The proto-oncogene c-myb plays an important role in regulation of cell growth and differentiation, and it is expressed at high levels in hematopoietic cells and many other types of cancers. However, the function of c-myb is not well known in OSCC. The present study aimed to reveal the function of c-myb and to test the alternation of cell growth and signaling by c-myb in OSCC. In this study, c-myb and dominant-negatibe myb(DNmyb) were expressed in an adenovirus-mediated gene delivery system to KB cells. The over-expressed c-myb brought increased cellular proliferation compared with control cells. However, DN-myb infected KB cells showed significant reduction of cell growth and enhanced induction of apoptosis to activate PARP and caspase 9. c-myb induced increase of IGF-I, -II and IGF-IR expressions while DN-myb down-regulated these expression. Activation of ERK and Akt/PKB pathway was shown only in c-myb transduced cells. These findings suggest that the role of c-myb in cell growth of oral cancer cells is partially mediated through the modulation of IGFs, ERK and Akt/PKB. From this results, DN-myb is strongly recommended as a curable gene for the treatment of c-myb dependent malignancies such as OSCC.

Oral squamous carcinoma (OSC) is the most common malignant neoplasm of the oral mucosa. Although the etiology of OSC is not fully understood, accumulated evidences indicate that the activation of proto-oncogenes and the inactivation of tumor suppressor genes underlie the disease development. An OSC cell line, YD-9 was newly established and characterized. However, the mutational analysis of p53 gene was not performed. Thus, in this study, the presence of mutation in the p53 gene was examined by amplification of exon-4 to -8 and subsequent DNA sequencing. Two point mutations were found in exon-4 and -6: A to G, resulting in amino acid change Tyr to Cys in exon-4, and C to G, resulting in amino acid change Gly to Arg in exon-6, respectively. Any mutation was not found in the exon-5, -7 and -8. The presented results would contribute to basic research to understand the biological mechanism of OSC using YD-9 cells.

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

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