The fruit of Kochia scoparia Scharder is traditionally used as a medicinal ingredient to treat allergic skin diseases and inflammatory diseases in China, Japan and Korea. Recently, several studies reported that K. scoparia had potential for the cytotoxicity of human cancer cells. To investigate the anti-cancer effect of K. scoparia on oral cancer and to determine the specific type of cell death induced by MEKS treatment. We investigated the anti-cancer effects of K. scoparia, methanol extract (MEKS) in HSC4 human oral cancer cells. We examined the effects of MEKS on the proliferation rate, cell cycle arrest, 7-AAD-ANNEXIN V double stain, reactive oxygen species (ROS) generation and activation of apoptosis and necroptosis-associated proteins in HSC4 cells. MTT assay results demonstrated that MEKS decreased the proliferation rates of HSC4 cells in a dose-dependent manner with an IC50 value of 45.3 μg/ml. MEKS at 50 μg/ml significantly increased the sub-G1 DNA contents of HSC4 cells to 84.8%, versus untreated cells. However, the activation of apoptosis-associated proteins such as cleaved caspase 3, cleaved caspase 8, cleaved caspase 9 and cleaved Poly (ADP-ribose) polymerase (PARP) did not detect. The level of Bax protein markedly increased in MEKS-treated HSC4 cells. In addition, the cell viability of the DPQ pre-treated HSC4 cells with MEKS treatment was significantly greater than that of MEKS treated-cells. These results suggest that MEKS inhibits cell proliferation and induces necroptosis in oral cancer cells and that MEKS may have potential chemotherapeutic value for the treatment of human oral cancer.

This study was performed to investigate the correlation between the primary squamous cell carcinoma in oral cavity (POSCC) and paired metastatic oral squamous cell carcinoma in cervical lymph node (MOSCC) via immunohistochemical staining with Ki-67 and p53. The subjects included ten patients (20 specimens) who were diagnosed with OSCC with metastatic lymph nodes from 2010 to 2015 and surgically treated involving neck dissection in Kyungpook National University Hospital. Twenty specimens were stained immunohistochemically with Ki-67 and p53. The degrees of immunostaining by Ki-67 and p53 was evaluated as 0 (no positive cells), weak (1~25% positive cells), moderate (26-50% positive cells) and strong (>50% positive cells). Despite the strong tendency, there was no statistically significant result between expressions of Ki-67 and p53 in POSCC or MOSCC. We found that high expression of Ki-67 was significantly correlated with poor degree of differentiation. Our results suggest that expression of Ki-67 may be a predictable factor for degree of differentiation of POSCC and MOSCC.

OSCC is currently the most common malignancy of the head and neck, affecting tens of thousands of patients per year worldwide. Natural flavonoids from plants are potential sources for novel anti-cancer drugs. Icariin is the active ingredient of flavonol glycoside, which is derived from the medical plant Herba Epimedii. A metabolite of icariin, icariside II exhibits a variety of pharmacological actions, including anti-rheumatic, anti-depressant, cardiovascular protective, and immunomodulatory functions. However, the exact mechanism causing the apoptosis-inducing effect of icariside II in OSCC is still not fully understood. In the present study, we assessed the anti-cancer effect of icariside II in OSCC cell lines by measuring its effect on cell viability, cell proliferation, and mitochondria membrane potential (MMP). Icariside II treatment of OSCC cells resulted in a dose- and time-dependent decrease in cell viability. Hoechst staining indicated apoptosis in icariside II-treated HSC cells. Icariside II inhibited cell proliferation and induced apoptosis in HSC cells, with significant increases in all present parameters in HSC-4 cells. The results clearly suggested that icariside II induced apoptosis via activation of intrinsic pathways and caspase cascades in HSC-4 cell lines. The collective findings of the study suggested that Icariside II is a potential treatment for OSCC; in addition, the data could provide a basis for the development of a novel anti-cancer strategy.

Human DEK gene on chromosome 6p encodes a 43kD nuclear phospoprotein that was originally identified as part of a fusion protein found in a subset of acute myeloid leukemia carrying a t(6;9) translocation. Although DEK upregulation has been described in a number of human malignancies and was significantly associated with high histologic grade, lymph node metastasis and/or advanced clinical stage, no previous report has evaluated the expression of DEK protein and its clinical significance in oral squamous cell carcinoma (OSCC). Our aims were to determine DEK expression in tissue samples of normal oral mucosa and OSCC by immunohistochemistry, to analyze the correlation between DEK expression and clinicopathological parameters, and to evaluate the value of DEK as a prognostic marker for patient’s survival. Ten normal oral mucosa, 10 epithelial dysplasia, and 60 OSCC samples were studied by immunohistochemistry. DEK expression tended to increase through the full thickness of epithelium in the dysplastic mucosa when compared with those in normal oral mucosa. High expression of DEK protein (score ≥ 2) was found in 68.3% of OSCC cases. Statistical analysis revealed that DEK overexpression in OSCC was positively correlated with high histologic grade (p=0.001), lymph node metastasis (p=0.003), and advanced clinical stage (p=0.039). In the Kaplan-Meier survival analysis, DEK overexpression was significantly associated with decreased overall survival in patients with OSCC (p=0.019). Our results suggest that DEK overexpression may be a reliable marker to predict the clinical outcome in OSCC.

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.

Many researchers have been reported recent trends for rising incidence particularly related to cancer of the tongue in young people. The purpose of this study is to exam the changes of OSCC(oral squamous cell carcinoma) incidence distribution. After Institutional Review Board approval, 1,345cases of OSCC were reviewed from 1993 to 2012 at the department of Oral Pathology at the University of Seoul National Dental Hospital with respect to gender, age, locations. As the results, the most prevalence ages were between 45-69 years old(mean:59.68). There is a noticeable incidence on the eighth decade of life. There are no changes of incidence distribution on se (male:female=69:31) at each year. The most common incidence area was tongue at 1993 but the mandible was the most popular site after 2001. Comparing the incidence rates of the predilection sites over and under 44 years old patients, among patients under 44 years old showed the tongue is the common site steadily. On the other hand, there is a significant increase on the mandible in the over 44 years old patients. In conclusion, this study showed similar results on overall sex distribution and median ages. The trends of locations over 20 years have been increased on the old adult’s mandible. Also, tongue is the common site for the young adults constantly. There are needs for further studies to elucidate the behind etiology. In addition, the true impact of young age on OSCC clinical behavior will remain difficult to determine unless intranational and multi-institutional databases will be combined.

Oral squamous cell carcinoma (OSCC), is the most common malignancy of oral cavity, and the sixth most frequently diagnosed cancer worldwide. This tumor type is associated with poor prognosis, and most OSCC patients are diagnosed after the cancer has reached an advanced stage. The over expression of NF-κB p65 has been associated with OSCC progression and lymph node metastasis. Hence, the present study analyzed the expression of NF-κB p65 in OSCC from Korean patients. Immunohistochemistry for NF-κB p65 was performed using 12 normal oral mucosas (NOM), 16 oral leukoplakia (with/without dysplasia), and 58 OSCC patients samples. Immunoreactivity was semi-quantitatively scored and the correlation between the expression of NF-κB p65 and clinicopathological parameters of OSCC patients was analyzed. Immunohistochemical staining revealed that NF-κB p65 expression level increased in oral leukoplakia with dysplasia and OSCC. Moreover, the immunoexpression of NF-κB p65 appeared to be associated with age, recurrence, TNM stage, and lymph node metastasis in OSCC patients (p<0.05). These results indicated that NF-κB p65 can play a role as oncogene in OSCC. Moreover, NF-κB p65 may play an important role in both oral carcinogenesis and OSCC patient outcome. It may be considered as another new malignant biomarker of OSCC.

Cinnamaldehyde is known to have the antitumor effects in vitro and in vivo. In this study, we showed a potent and irreversible cytotoxic activity of the Cinnamaldehyde derivative 2'-benzoyloxycinnamaldehyde (BCA) in human Squamous oral cell carcinoma cell, YD-10B. BCA induced YD-10B cell apoptosis in a dose-responsive manner. BCA-induced apoptosis was associated with corresponding increases in a series of key components in the mitochondria-mediated apoptosis pathways, followed by caspase cleavage and PARP activation. We also observed that BCA induced autophagy through Akt/mTOR pathway in YD-10B cells. BCA treatment increased LC3B-II expression, and induced the formation of autophagosomes and autophagic vacuoles. These experimental findings suggest that BCA is a potent inhibitor of cell proliferation in YD-10B cells and provide new insights about leading to the possible development of a new therapeutic agent.

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.

The molecular mechanisms of the carcinogenesis of oral squamous cell carcinomas (OSCCs) are highly variable and result in different features of tumor progression, i.e., local tissue destruction and metastasis to regional lymph nodes. A case of OSCC arising from proliferative verrucous leukoplakia (PVL) was analyzed for its protein expression profile by immunoprecipitation (IP) – high performance liquid chromatography (IP-HPLC) by using 72 antisera and comparing results with those of KB cells. OSCC arising from PVL showed stronger expressions of proteins associated with cell proliferation (MPM2, PCNA, eiF5A, DHS, DOHH), cell survival (pAKT, MDM2, survivin), matrix proteolysis (elaffin), tumor suppression (p16, p21, PTCH1), the WNT/β-catenin pathway (SHH, WNT1, APC, β-catenin, snail), proinflammation (TNFα), angiogenesis (HIF, CMG2, vWF), and cellular protection (HSP-70, FAK, caveolin) and of oncoproteins (STAT3, 14-3-3, K-RAS, PUMA, PIM1) and growth factors (EGFR, bFGF) than KB cells. On the other hand, KB cells showed stronger expressions of proteins associated with apoptosis (caspase-3, caspase-8, caspase-9, PARP, FAS, FASL, TGase-1, BCL2, BAD, BID, BAK, FLIP), matrix proteolysis (MMP-2, MMP-9), transcription signaling (NFkB, p38, E2F-1, HO-1), and tumor suppression (p53, RB1, PTEN) and of oncoproteins (DMBT1, CEA) and growth factor (TGF-β1, c-erbB2, VEGF) than OSCC arising from PVL. These data indicate the cells of OSCC arising from PVL are more resistant and more robust than KB cells. Furthermore, they suggest the oncogenic signalings of OSCC arising from PVL play important roles in the aggressive growth and rapid tumor metastasis to regional lymph nodes

Primary oral squamous cell carcinoma (OSCC) associated with dental osseointegrated implants is very rare. We experienced two patients who had received dental implant surgery before they were diagnosed with OSCC. We report these cases to emphasize the importance of differential diagnosis of malignant lesions associated with dental implants. Additionally, we also suggest that bone graft materials around implants can serve as a potential inducer of invasiveness in cancer cells.

Development of squamous cell carcinoma around dental implants is an uncommon clinical manifestation with only a few cases described in the literature. Recently, we observed primary squamous cell carcinoma that developed from leukoplakia around dental implants. We report this case to emphasize the importance of careful oral examination, for implant surgery has to be preceded by thorough evaluation of oral mucosal conditions.

Chronic inflammation is widely considered to predispose individuals to cancer. Microorganisms facilitate recruitment and activation of inflammatory cells and thus allow release of inflammatory mediators. These molecules can then promote accumulation of mutations, leading to tumor development in the host. Porphyromonas gingivalis, a pathogen causing chronic periodontitis, is detected in oral squamous cell carcinoma (OSCC) tissues. Considering a strong link between chronic inflammation and tumor development, functional consequences of P. gingivalis infection may include malignant transformation of the host cells. In this study, we monitored transcriptional changes induced by invasion of P. gingivalis in OSCC cells using microarrays. Our preliminary results suggest changes in a wide range of genes involved in inflammation, apoptosis and autophagy, tumor progression, and carcinogenesis. Further studies on molecular mechanisms underlying these changes will lay a useful foundation to elucidate the role of microorganism-related inflammation and for the development of preventive and therapeutic agents for oral cancer.

Inflammation functions as a double-edged sword against external stimulus. For instance, inflammation can have anti-cancer effect and simultaneously can play cancer-promoting factors. Recent studies have shown that cytokine plays an important role in tumor biology by influencing tumor growth, invasion and metastasis. We classify these cytokines by cancer type and review current knowledge of cytokines in terms of carcinogenesis. Here, we also focus on whether cytokines can act as biomarkers for early detection of oral squamous cell carcinoma (OSCC). This review will provide basis for further approach to study the role of cytokines in carcinogenesis and evaluating the possibilities of cytokines as biomarkers for cancer detection

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.

Oral squamous cell carcinoma (OSCC) is one of the most common carcinomas in the head and neck area. Bitter melon extraction (Momordicacharantia, BME) has been used as a functional food to prevent and treat human health related issues. It has recently been reported that BME inhibits breast cancer cell growth by arresting cell cycle and promoting apoptosis. In this study, we aimed at proving the inhibitory action of BME on OSCC proliferation. We used two OSCC cell lines, SCC4 and Ca9-22. Both cell lines were treated with different concentrations (1%, 2%, and 5%) of BME. Cell viability was examined by MTT assay. DNA condensation was visualized by immunofluorescence microscope to determine the signs of the cell apoptosis. Cell numbers were significantly decreased in a dose-dependent manner by bitter melon at concentration of 1% of BME on Ca9-22 cell line (P=0.001) but no significant effect on SCC4 (P=0.124) at the same concentration. 2% BME treatment of the Ca9-22 cell line induces chromatin condensation and DNA fragmentation after 20 hours. It seems that BME inhibits the proliferation of Ca9-22 cell line by inducing apoptosis. Thus, BME may be used as a dietary supplement for prevention of OSCC.

Tumor cells, especially in a malignant form, proliferate rapidly that blood supply within the tumors becomes limited, leading to a condition of insufficient oxygen supply. Such hypoxic condition is known to impair the viability of cancer cells, but it can also be a factor to facilitate the appearance of cells with a higher degree of malignancy. Indeed, the hypoxic condition created within malignant tumors may contribute to promoting their aggressive behaviors, including tumor invasion and metastasis, and to the development of resistance to various therapeutic modalities such as chemotherapy. Recently, microRNAs, a group of short RNA fragments consisting of 18 to 20 nucleotides, have been shown to participate in regulating genes important for cell survival, differentiation and apoptosis. Since their discovery, modulation of these microRNAs has been a focus of intensive studies with regard to their significance on gene regulation and various aspects of cell biology. In this study, we investigated hypoxia-induced alterations in microRNAs in oral squamous cell carcinoma (OSCC) cells and discussed consequential gene modulation and relevant cellular responses.

Metastasis consists of complex cascades and a lot of factors are involved in each step of metastasis. In recent studies, the role of epithelial-mesenchymal transition (EMT) in metastasis is suggested. EMT has a feature of epithelial cells conversing into mesenchymal cells morphologically and phenotypically, is a characteristic of malignant and metastatic cells in most cancer. The mesenchymal cells usually show more malignant phenotype, including invasion and metastasis. EMT can play an important role in metastasis of oral squamous cell carcinoma (OSCC). Although the role of Snail, slug, other transcriptional factors and E-cadherin are well known in human cancers, there are a few studies on N-cadherin and Twist expression in OSCC. The present study was aimed to analyze the expression of N-cadherin and Twist protein in OSCC from Korean patients. The immunohistochemical stain was performed using 58 primary OSCCs and 6 metastatic OSCCs, and the correlation between the expression of these proteins and clinicopathological parameters of OSCC patients was analyzed. The expression rate of high expression of N-cadherin was observed in 70.4% and Twist in 87.3% of OSCC. The expression of N-cadherin in metastatic OSCC increased than in corresponding primary OSCC (p<0.05). The spearman correlation coefficiency between N-cadherin and Twist was calculated as 0.228. The clinical factors such as lymph node metastasis and survival showed statistically significant correlation between N-cadherin expression. The expression of Twist was correlated with recurrence. In conclusion, the authors suggest that N-cadherin may play an important role in malignant behaviour of OSCC and can be considered as prognostic indicator of OSCC.

Oral squamous cell carcinoma (OSCC) develops through multistep process, that is, from normal mucosa to hyperplastic area and progressed to dysplasia, carcinoma in situ, and finally to invasive carcinoma. The purpose of this study is to investigate the histological types of the transitional area from normal oral mucosa to invasive carcinoma for the baseline data to search intermediate end point markers for early detection of OSCC. For this purpose, we reviewed the 85 patients who were diagnosed as OSCC in the Department of Oral Pathology, Yonsei University College of Dentistry, from 2002 to 2008. We classified these histopathologic findings by light-microscopy, according to the histologic pattern of transitional areas. As results, stepwise transformation from normal oral mucosa, to dysplasia and to OSCC was shown in 47 patients. Intermittent lesions were seen in 16 patients, in which normal oral mucosa, dysplasia, and OSCC were alternately arranged. Twenty two patients showed abruptly transformed to OSCC from normal oral mucosa. These preliminary data will be used for searching biomarkers for early detection of OSCC.

Cruciferous vegetables including diindolylmethane (DIM) have been shown to have anticancer activity. Especially, DIM-pPhBr and DIM-pPhF used in this study was reported to have more effective and less toxic effects than DIM. However, there is no report presenting their anti-tumorigenic activity in oral cancer. In the present study, we examined the effects of DIM-pPhBr and DIM-pPhF on the cell proliferation and apoptosis in KB human oral cancer cells. DIM-pPhBr and DIM-pPhF decreased cell proliferation and induced apoptosis evidenced by western blot analysis, DAPI staining and sub-G1 population. This provides the first evidence that DIM-pPhBr and DIM-pPhF originating from cruciferous vegetables induce apoptotic cell death in human oral cancer cells to inhibit cancer cell proliferation.