Chronic inflammation has long been considered as an important contributing factor to the development of malignant tumors in various tissues. In this study, we aimed to investigate a potential association between chronic periodontitis, a representative inflammatory disease in the oral cavity, and oral squamous cell carcinoma (OSCC), the most common form of malignant tumors in the oral cavity. A retrospective study was designed to include the cases and controls, each of which consisted of patients first diagnosed with OSCC and temporomandibular disorders, respectively. The existence or a history of periodontal disease was quantitatively estimated based upon the level of alveolar bone loss (ABL) from panoramic radiographs in these groups. Unlike other covariates, including LDH, WBC count and hemoglobin, the levels of ABL measured at three independent regions (second premolar and first/second molar) were significantly higher in the OSCC group, regardless of the patients’age in most cases. Our results thus support the hypothesis that chronic periodontitis, represented by significant ABL, is an important and clinically relevant factor potentially associated with the development of OSCC.

Neuromedin B (NMB) acts as a growth factor or a morphogen and plays a role in cancer progression. Indeed, the NMB receptor (NMB-R) is overexpressed in different types of tumors. In our current study, we investigated the involvement of NMB-R in the proliferation of oral cancer cells. Human oral squamous cell carcinoma (SCC) and human oral cancer cells, SCC-25 cells were found to be NMB-R-positive. The NMB-R antagonist PD168368 inhibited the proliferation of SCC-25 cells and reduced their colony formation capacity. We also found that PD168368 induced the cell cycle arrest and apoptosis of SCC-25 cells in a dose-/time-dependent manner. Overall, this antitumor activity of PD168368 in human oral cancer cells suggests that NMB-R is a potential target for the future prevention and treatment of human cancers.

Dibenzylideneacetone (DBA), an analogue of curcumin has been shown to have anti-cancer activity in a variety of tumor cell lines. However, the anti-cancer activity of DBA and its molecular mechanism in HN22 oral cancer cell line have not been fully explored. The effects of DBA on anti-proliferative and apoptotic activity were evaluated by the trypan blue exclusion assay, 4’-6-diamidino-2-phenylindole (DAPI) staining, Western blot analysis, and reverse transcriptase-polymerase chain Reaction (RT-PCR). Our data showed that the treatment of DBA to HN22 cells exerted anti-proliferative and apoptotic activities and the activity was accompanied by a decrease in Sp1 protein, Sp1 mRNA and its promoter activity. DBA also reduced the expression level of Sp1 protein and caused apoptotic cell death in HN22 cells simultaneouly. Phosphorylation of ERK and JNK were regulated by DBA whereas phosphorylation of p38 was not altered. Overall, our results suggest that the regulation of Sp1 activities and ERK/JNK are involved in DBA-induced apoptosis and DBA can be a promising anticancer drug candidate for the treatment of oral cancer.

The presence of distinct bacterial species is found to be dependent on age, diet, and disease. We compared the detection rate of several oral bacterial strains in a cohort of 36 subjects including healthy volunteers, periodontal patients, and oral cancer patients. Gargling samples were obtained from these subjects from which DNA was then extracted. Specific primers for 29 bacterial species were used for PCR detection. In the oral cancer patients, Capnocytophaga ochracea, Gemella morbillorum, and Streptococcus salivarius were detected more frequently compared with the healthy volunteers and periodontitis patients. Fusobacterium nucleatum/ polymorphym and Prevotella nigrescens were significantly less prevalent in oral cancer patients than the other groups. In periodontitis patients, Porphyromonas gingivalis and Treponema denticola were more frequently found compared with the healthy volunteers. In the healthy volunteer group, Peptostreptococcus anaerobius was more frequently found than the other groups. The detection rate of several oral bacterial species was thus found to differ between healthy volunteers, periodontitis patients and oral cancer patients.

Bile acids and synthetic bile acid derivatives induce apop-tosis in various kinds of cancer cells and thus have anti-cancer properties. Recently, it has been suggested that autop-hagy may play an important role in cancer therapy. How-ever, few data are available regarding the role of autophagy in oral cancers and there have been no reports of autophagic cell death in OSCCs (oral squamous cell carcinoma cells) in-duced by HS-1200, a synthetic bile acid derivative. We thus examine whether HS-1200 modulates autophagy in OSCCs. Our findings indicate that HS-1200 has anticancer effects in OSCCs, and we observed in these cells that autophagic vacuoles were visible by monodansylcadaverine (MDC)and acridine orange staining. When we analyzed HS-1200-treated OSCC cells for the presence of biochemical markers, we observed that this treatment directly affects the conversion of LC-3II, degradation of p62/SQSTM1 and full-length beclin-1, clea-vage of ATG5-12 and the activation of caspase. An autop-hagy inhibitor suppressed HS-1200-induced cell death in OSCCs, confirming that autophagy acts as a pro-death signal in these cells. Furthermore, HS-1200 shows anticancer acti-vity against OSCCs via both autophagy and apoptosis. Our current findings suggest that HS-1200 may potentially cont-ribute to oral cancer treatment and thus provide useful infor-mation for the future development of a new therapeutic agent.

Topoisomerases are essential enzymes involved in all processes of DNA metabolism, and their inhibitors have been identified as potential anti-cancer agents. The present study examined the effect of linoelaidic acid (C18 polyunsaturated fatty) compounds derived from Gardenia jasminoids Ellis extract on the activity of eukaryotic topoisomerases inhibition. The present study identified linoelaidic acid compounds using open column fraction, HPLC, NMR and LC/MS which have effects on cell death in oral cancer cell line, FaDu, but not in immortalized normal cell line, HaCaT. Subsequent studies revealed linoelaidic acid-induced autophagy through LC3 activation. Finally, its inhibition of topoisomerase I and selectively induction of oral cancer cell death possibly implies that linoelaidic acid can be a role as potenial agents in the prevention and therapy of oral cancer.

MicroRNAs (miRNAs) are small non-coding RNAs that mediate gene expression at the post-transcriptional level by degrading or repressing targeted mRNAs. These molecules are about 21-25 nucleotides in length and exert their effects by binding to partially complementary sites in mRNAs, predominantly in the 3'-untranslated region (3'-UTR). Recent evidence has demonstrated that miRNAs can function as oncogenes or tumor suppressors through the modulation of multiple oncogenic cellular processes in cancer development, including initiation, cell proliferation, apoptosis, invasion and metastasis. In our present study, we examined the expression profile of miRNAs related to oral cancer cell growth inhibition using normal human oral keratinocytes (NHOK) and YD-38 human oral cancer cells. By miRNA microassay analysis, 40 and 31 miRNAs among the 1,769 examined were found to be up- and down-regulated in YD-38 cells compared with NHOK cells, respectively. Using qRT-PCR analysis, the expression levels of miR-30a and miR-1246 were found to be increased in YD-38 cells compared with NHOK cells, whereas miR-203 and miR-125a were observed to be decreased. Importantly, the overexpression of miR-203 and miR-125a significantly inhibited the growth of YD-38 cells. This finding and the microarray data indicate the involvement of specific miRNAs in the development and progression of oral cancer.

Runt-related transcription factor (RUNX) 3 is well known as a developmental regulators, as well as candidate tumor suppressor gene in human breast cancer, gastric cancer, esophageal cancer, and so on. The present study was aimed to analyze the expression of RUNX3 protein in oral squamous cell carcinoma (OSCC) from Korean patients. The immunohistochemical stain was performed with 14 normal oral mucosa (NOM) and 25 OSCCs, and statistical analysis was carried out to find out the correlation between the expression of RUNX and clinicopathological parameters of OSCC patients. In OSCC, the expression of RUNX3 protein was found to increase more than in NOM. Moreover, in the univariate correlation analysis, the gender, regional lymph node metastasis, and histopathologic differentiation of OSCC patients were positively correlated with the expression of RUNX3 (p<0.05). These results indicate that RUNX3 can play a role as an oncogene in OSCC, in contrast to some reports on RUNX3 in other human cancers. In addition, RUNX3 may be considered as new malignant biomarker of OSCC.

Cancer cells are often found in an ischemic condition due to the rapid outgrowth of their vascular supply, and these cells are expected to develop an increased potential for local invasive growth. Since the first steps are characterized by increased motility and invasiveness, expression of molecules involved in cellular adhesion to extracellular matrix (ECM) is increased in the process of cancer cell invasion and metastasis. In this work we explored the molecular characteristics and its regulatory mechanism of hypoxic oral squamous cell carcinoma (OSCC) cells. Our experiment identified that hypoxia increases α5 integrin protein levels through phosphoinositide 3-kinase (PI3K)/Akt pathway in OSCC cells.

Angelica decursiva has been used in Korean traditional medicine as an antitussive, an analgesic, an antipyretic and a cough remedy. However, its anti-cancer properties have not yet been well defined. In our current study, we report the cytotoxic activity and the mechanism of cell death induced by ethanol extracts of Angelica decursiva (EEAD) against the human oral cancer cell line, KB. Treatment of KB cells with EEAD induced apoptotic cell death in both a dose- and time-dependent manner as determined by MTT assay and DNA fragmentation. However, no cytotoxic effects of EEAD against human normal oral keratinocytes (HNOK) were evident. By western blot analysis, we found that apoptosis in KB cells is associated with a decrease in procaspase-7 and -9. In addition, the activation of caspase-7 was detectable in living KB cells by fluorescence microscopy. These results suggest that EEAD exhibits anti-cancer activity in KB cells via apoptosis and thus has potential as an anticancer agent in future drug development strategies.

Oral squamous cell carcinoma (OSCC) has been a focus of cancer prevention studies due to the fact that it occurs by a multistep process and that a precancerous lesion in the oral mucosa is easily accessible. The present study was aimed at developing an optical detection system using autofluorescence spectrum measurements for the early detection of oral cancer. The optical detection system was designed to use an excitation wavelength of 337 nm emanating from a Xenon lamp. Precancerous and cancerous lesions were created in the hamster buccal pouch by treatment with 7,12-dimethylbenz[a]anthracene (DMBA). Four groups of five hamsters each were used in this experiment. The right buccal pouch was treated with 0.5% DMBA to induce carcinogenesis while the left buccal pouch was treated with mineral oil as a control. The autofluorescence of both buccal pouches was measured weekly. A difference in the excitation pattern between the normal and the carcinogen-treated tissue was noticed after three weeks. Specifically, the intensity of the autofluorescence spectrum in the DMBA-treated buccal pouch was increased at wavelengths between 400 and 450 nm. The results of the autofluorescence measurements were compared to histological findings and show that the intensity of the autofluorescence increased along with the stage of epithelial dysplasia. Based on the fact that one of the autofluorophores in this tissue is NADH, we measured the fluorescence at the 450-nm NADH wavelength to conclude that the increased autofluorescence in the dysplastic areas may be caused by NADH. Based on these data, we suggest that autofluorescence optical methods are a useful tool for the early detection of oral cancer.

PDT is an established cancer treatment modality. This can be attributed to the attractive basic concept of PDT; Combination of two therapeutic agents, a photosensitizing drug and light, which are relatively harmless by themselves but when combined, cause more or less selective tumor destruction. Hematoporphyrin-derived photosensitizers are known to be stable and highly efficient. In this study, we conducted a series of experiments to develop light-induced anticancer drugs against oral cancer cells. We tested the cytotoxicity of photodin by MTT assay and observed cell death pattern (apoptosis or necrosis) by hoechst 33342 and propidium iodide staining methods after PDT. IC50 value of photodin was 0.65 ug/ml. At higher doses of photodin ( > 7.8 ug/ml), cancer cells died exclusively from necrosis after PDT. By contrast, at IC50 value, photodin induced cancer cell to undergo apoptotic cell death. The induction begins approximately 6 hours after PDT. We investigated intracellular localization of photodin by oral cancer cell via confocal laser scanning microscopy. Oral cancer cells dual-stained with photodin and organelle-specific fluorescence probes (Mitotracker, Lysotracker, ER-Tracker) revealed that an intracellular fluorescence distribution was restricted to cytoplasmic compartments with no detectable fluorescence in the nucleus. Confocal images of cells containing photodin were overlapped with the mitochondria-specific fluorescence probe images of the same cells. These results demonstrated that photodin may play the role of a photosensitizer for oral squamous cancer cells without swelling and inflammation. Therefore, photodin-based PDT is a suitable treatment for oral cavity carcinoma patients.

We conducted a series of in vitro experiments to evaluate the anticancer effect of photodynamic therapy using hypericin and 532㎚ DPSS (diode pumped solid state laser). The cultured KB cells were treated with serial concentrations of hypericin ranging from 0.01㎍/㎖ to 5㎍/㎖ (two-fold dilution) with variable laser dosage (10J, 20J, 30J). The cell viability was evaluated by MTT assay. The type of cell death was detected by fluorescent microscope using Hoechst 33342 / PI (propidium iodide) stain methods. In this study, IC50 value with hypericin-mediated PDT with 10J DPSS laser was 35 ng/ml. The maximum cytotoxicity with Photofrin II-based PDT was observed at high drug concentrations(> 90 ng/ml) independent with laser dose. And the in vitro PDT effects depended on the laser dose and drug concentrations were displayed by the difference in the type of cell death, namely apoptosis or necrosis. According to this result, the hypericin based photodynamic therapy with DPSS laser was effective photodynamic therapy.

Eukaryotic translation initiation factor 5A (eIF-5A) is essential for proliferation of eukaryotic cells, andwas identified as diagnotic marker in cervical intraepithelial neoplasia, cervical and endometrial cancer, but relatively little is known about thein vivo and in vitro expression patterns of eIF-5A in oral premalignant and malignant lesions mirror the expression levels observed in vitro with cells derived from normal oral mucosa, immortalized oral keratinocytes (IHOK) and primary and metastatic oral squamous cell carcinoma (OSCC). We used an oral squamous cell carcinoma (OSCC) progression model composed of cell lines and tissue specimens to characterize expression patterns by Western blotting and immunohistochemistry. eIF-5A and PCNA levels are elevated in IHOKand primary and metastatic OSCC cella as compatred to normal human oral keraitinocytes. eIF5A and PCNA expression was l imited to basal cells of normal oral mucosa. eIF-5A and PCNA expression is increased in dysplastic epithelium spreading to more superficial layers, and its expression levels correlated significantly with the degree of dysplasia. Well and moderately differentiated OSCC showed strong expression of eIF-5A and PCNA. These results suggest that upregulated expression of eIF-5A seems to be an important epigenetic alteration that accompanies oral carcinogenic progression, and eIF-5A could be used as an biomarker for oral premalignat lesion or squamous cell carcinoma

Considering the great potential of iron chelators at inhibiting the proliferation of tumor cells, in order to determine the molecular and biological basis for the effects of iron chelator in oral cancer, we investigated the effects of iron chelator, desferrioxamine (DFO), on the gene profiling analysis of immortalized human oral keratinocytes (IHOK), and oral cancer cells (HN12), using the cDNA microarray. We identified 46 clones cDNA exhibiting more than 2 fold overexpression in DFO treated IHOK and HN12 cells, and 94 cDNA reveal more than 2 fold down-regulated expression. Examination of gene expression that differs between DFO treated vs. control IHOK and HN12 cells apprear to be related to : cell cycle regulator, cell growth and apoptosis, signal transduction and stress. p21 for cell cell cycle factor was upregualted, and cyclin-cdk gene was decreased expression, so we observed cell cycle arrest in DFO treated IHOK and HN12 cells. In tumor growth, we have identified downregulation of hemidesmosomal protein (bullous pemphigoid antigen 1) and epiregulin expression in DFO treated IHOK and oral cancer cells. Signal transducers including mitogen-activated protein kinase-activated protein kinase 5, serine/thereonine kinase 6 were downregulated with DFO treated cells, suggesting the DFO regulates the p38 MAP kianse pathway in immortalized and maignant oral keratincytes. In conclusion, we have demonstrated the high-throughput utility of cDNA array hybridization in parallel to the gene expression analysis to identify genes that are expressed differentially in DFO treated with immortalized and malignant oral keratinocytes. The differentially expressed genes identified here should be informative in DFO-induced anti-cancer effects.

Inhibition of proteasome activity may reduce many types of cancer, so it's pathway is effective in cancer as well as in clinical fields. Here the author has carried out experiment targeting on the elevation of apoptosis in oral cancer cells by combination of proteasome inhibitor, lactacystin, and DNA replication inhibitor, etoposide. The growth of KB cells was measured by MTT methods and apoptosis was analyzed by DNA fragmentation and Hochest nucleus staining. The proteasome activity was analyzed by fluorescent tagged peptide and cellular protein expression was detected by Western hybridization. Though lactacystin and etoposide inhibited KB cell growth alone, but low combined doses inhibited cell growth more strongly and induced apoptosis. The proteasome activity was also seriously inhibited by the combination of both chemicals. Tumor suppressor proteins and apoptosis inducing proteins were highly increased under the combination of both chemicals. From above studies we can conclude that proteasome inhibitors may be used for the treatment of oral cancer and proteasome inhibitors with DNA replication inhibitors may be effective in clinical trials of oral cancer.

PDT is an established cancer treatment modality, This can be attri buted to the attractive basic concept of PDT; the combination 0 1' two therapeutic agents , a photosensiti zing drug and light, which are relatively harmless by themselves but combined ultimately cause more 0 1' less selective tumor destruction, The bacteri ochlorophyll - derivatived photosensitizer s are known to be s tabl e and highly effïcient, ln thí s study, we conducted a seríes of experiments to develope the light induced anticancer drugs against oral cancer cell , We tested the cytotoxi city of the hydroxybacteriochlorine by MTT assay and observed the cell death pattern(apoptosis or necrosis) after PDT by hoechst 33342 and propidium iodide s taining methods, lC50 value of the hydroxybac teriochlorine was 3 1 ， 3 ng/ n띠， At higher doses of hydroxybacteriochlori ne () 60ng/ rnQ) , cancer cells died exclus ively by necrosis after PDT By contrast, at lC50 value, hydroxybacteri ochlorine induced cancer cell to undergo apop totic c e ll death, The induct ion begins approximately 6 hours a fter PDT, We inves tigates intrace l1 ular localization of hydroxybacte riochlorine by oral cancel‘ cell via confocal laser scanning mi croscopy, Oral cancer cells dua l-stained with hydrox ybacteriochlo1' ine and organelle-specific flu orescence probes (Mi totracker , Lysotracke1', ER- Tracker) revealed an intracellular f1 uores c ence dis tribution restricted to cyt oplasmic compartments with no det ectable fluorescence in the nucleus, Confocal im ages of cells containing hydroxybacte1'iochl orine were never overla p to mitochondria, lysosome . endoplasmic reticulum when digita lly overla pped with tqe organel1e-specific f1 uorescence p1'obe images o{' the same cells These results demons trated tha t the hydroxybacte1' iochlorine may have a fun ction as a photosensitizer and cytotoxicity hydroxy bacteriochlo1'ine for o1'al cancer cell is more sensitive than head & neck cancer cell 0 1' ce1'vical cancer cell Therefore PDT using hydroxybacte1'iochl orine is suita ble treatment for oral cavity carcinoma patients

Sulfur is commonly used in Asia as a n herba l medicine to treat infl ammation and cancel‘. and potent chemopreventive effects have been demonstrated in various in vivo and in vitromodels for sulfur-containing compounds found in naturally occun‘ ing products. Here, we report the growth inhibitory and apoptosis-related effects of a newly developedhigh- purity eclible sulfur (ES) on immortali zecl human oral keratinocytes (IHOKs) and on oral cancer cells representing two stages of oral can cer (HN4‘ HN12) basecl on an 3-(4. 5-Dimethylt hiazol-2-yl)-2.5-cliphenyl tetrazolium bromide (MTT) a ssay, Western blotting, cell cycle analysis, ancl nuclear staining. The puri ty of the ES used in th is s tucly was verified by high performance liquid chromat ography (HPLC) , amino acid analysis and energy di spersive spectroscopy (EDS). ES inhibitecl the proliferation of immor talized and malignant oral kerati nocytes in a dose- and time-dependent manner FITC-Annexin V staining. DNA fragmentation testing. and Hoechst 33258 staining revealed that ES inhibits cell growth via apoptosis . ES blocked cell-cycle progression at the sub- Gl phase, with decreased expression 0 1' cyclins Dl, D2, and E, and t heir activating partners cdk2, cdk4, and cdk6‘ and a concomitant induction of p53 and p21/WAF1. Furthermore, ES treatment increased the cytosolic level of cytochrome c a nd resulted in caspase-3 activation‘ and thi s effect was correlated wi th Bax up- regulation and Bcl-2 down- regulation Taken together, these clata suggest that ES is a potential chemopreventive and chemotherapeutic agent for oral cancel

Epithelial-mesenchymal transition(EMT) plays a pivotal role in the convers ion of earl y s tage tumors into invasive malignancies‘ and has been shown to be regulated hy the transcri ptional factor. Snail. Recent ly‘ actlvatlon of the phosphatidylinositol 3' kinase (PI3K)/따<:T axis is emerging as a centra l feature of EMT‘ However. it is unclear whether the phosphorylation of AKT regulate the expl'ession of s nail in ora l cancer cell underwent EMT. T。 investigate a role of p-AKT in EMT, we assessed the effects of inhibi ting p-AK1' activity in oral squamous can cer cells(KOSCC-25B) using PIAs, structurally modified phosphatidyli nositol ether lipid analogues(P1As) . PIAs de creased phosphorylation of c-Jun N-terrninal Kinase(JNK) and increased phosphorylation of glycogen synthase kinase 3beta(GSK-3beta). Inhibition of p-AKT ir빼ce d down regulation of Snail and Twist. but Sip1 regulated independent of p-AKT inhibition. Also inhibi tion of p-AKT dec reased cell migration and invas ion. Therefore our results implicate that p-AKT may contribute to the translocalization of sna il in the EMT associated with canceJ cell rnigration and invasion

Sulfur is commonly used in Asia as an herbal medicine to treat inflammation and cancer , a nd potent chemo preventi ve effects have been demons tra ted in various in vivo and in vitromodels for s ul fur-containing compounds found in natura l1y occurring product s. Here, we 1'eport the growth inhibitory and apoptosis-related effects of a n ewly developedhigh-puri ty edible sulfur(ES) on immo1'tali zed human o1'al ke1'atinocytes(IHOKs) and on oral cancer cells representing two stages of oral cancer (HN4‘ HN12) based on an 3-(4,5-Dimethylthiazol-2-yl) -2.5- diphe n yltetrazolium bromide(MTI) assay, Western blotting, cell cycle analysis, and nuclear staining. The puri ty of the ES used in thi s study was ve1'ified by high performance liquid chromatog1'aphy (HPLC) , ami no acid analysis and energy dispersive spectroscopy(EDS). ES inhibited the prolife1'ation of imrnortalized and ma lig nant o1'al kerati nocytes in a dose- and time-dependent manne1' FITC-Annex.in V staining, DNA fragmentation t esting. and Hoechst 33258 s taining revealed that ES inhibits cell growth via apoptosis. ES bl ocked cell-cycle prog1'ession at t he sub-Gl phase‘ wi th decreased expression of cyclins Dl, D2‘ and E, and their activating partn ers cdk2‘ cdk4‘ and cdkfì, and a concomitant induction of p53 and p21/WAF1. Furthe1'more, ES treatment in creased the cytosolic level of cytochrome c and resulted in caspase- 3 activation‘ and thi s effect was co1'1'elated with Bax up-regulation and Bcl-2 down-1'egulation Taken together‘ these data suggest that ES is a potential chemopreventive and chemotherapeut ic agent fo r oral ca ncer