Oral squamous cell carcinoma (OSCC) is the most common type of oral malignancy. Numerous therapies have been proposed for its cure. Research is continually being conducted to develop new forms of treatment as current therapies are associated with numerous side-effects. Luteolin, a common dietary flavonoid, has been demonstrated to possess strong anti-cancer activity against various human cancer cell lines. Nevertheless, research into luteolin-based anticancer activity against oral cancer remains scarce. Thus, the objective of this study was to assess the effect of luteolin as an anti-cancer agent. After treatment with luteolin, Ca9-22 and CAL-27 oral cancer cells showed condensed nuclei and enhanced apoptotic rate with evidence of mitochondria-mediated apoptosis. Epithelial-mesenchymal transition (EMT) is closely related to tumor migration and invasion. Luteolin suppressed cancer cell invasion and migration in the current study. Elevated expression of E-cadherin, an adherens junction protein, was evident in both cell lines after luteolin treatment. Luteolin also significantly inhibited transcription factors (i.e., N-cadherin, Slug, Snail, Twist, and ZEB-1) that regulated expression of tumor suppressors such as E-cadherin based on Western blot analysis and quantitative PCR. Thus, luteolin could induce mitochondrial apoptosis and inhibit cancer cell invasion and migration by suppressing EMT-induced transcription factors.

Codium fragile (Suringar) Hariot is an edible green seaweed that belong to the Codiaceae family and has been used in Oriental medicine for the treatment of enterobiasis, dropsy, and dysuria. Methanol extract of codium fragile has anti-oxidant, anti-inflammatory and anti-cancer properties, although the anti-cancer effect on oral cancer has not yet been reported. In this study, we investigated the anti-cancer activity and the mechanism of cell death by methanol extracts of Codium fragile (MeCF) on human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that MeCF inhibits cell viability in a dose-dependent manner, and markedly induced apoptosis, as determined by the MTT assay, Live/Dead assay, and DAPI stain. In addition, MeCF induced the proteolytic cleavage of procaspase -3, -7, -9 and poly(ADP-ribose) polymerase(PARP), and upregulated or downregulated the expression of mitochondrial-apoptosis factor, Bax(pro-apoptotic factor), and Bcl-2(anti-apoptotic factor), . Futhermore, MeCF induced a cell cycle arrest at the G1/S phase through suppressing the expression of the cell cycle cascade proteins, p21, CDK4, CyclinD1, and phospho-Rb. Taken together, these results indicated that MeCF inhibits cell growth, and this inhibition is mediated by caspase- and mitochondrial-dependent apoptotic pathways through cell cycle arrest at the G1/S phase in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, methanol extracts of Codium fragile can be provided as a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.

The role of CXCR7, a seven-transmembrane G-protein coupled chemokine receptor, which binds with high affinity to chemokine CXCL11 and CXCL12 in oral cancer cells and the effect of transient CXCR7-downregulation on proliferation and migration of oral squamous cell carcinoma (OSCC) cells have not been reported. The aim of the present study was to evaluate the effects of CXCR7 on an OSCC cell line. In this study, we down-regulated CXCR7 in the KOSCC25B OSCC cell line by siRNA. In vitro cell proliferation and migration assays were used to investigate the effect of CXCR7- downregulation on cell proliferation and migration in si.KOSCC25B cells. The CXCR7 down-regulated OSCC cells grew significantly slower than the negative control siRNA transfected KOSCC25B cells (p<0.05). Additionally, migration of si.KOSCC25B cells decreased significantly compared with non-transfected KOSCC25B cells (p<0.007). These results suggest that down-regulation of CXCR7 induces anti-proliferative and anti-migratory effects in OSCC, and that CXCR7 may be a useful target molecule for the treatment of OSCC.

Radiotherapy (RT) is a mainstay in the treatment of head and neck squamous cell carcinoma (HNSCC). For locally advanced HCSCC, concurrent chemoradiotherapy (CCRT) benefits HCSCC patients in terms of better survival and loco-regional control. In this study, we evaluated changes in oral microbiota in patients, who received CCRT for head and neck cancer. Oral rinsed samples were weekly collected before and during CCRT and at 4 weeks following treatment from HNSCC patients, who had received 70 Gy of radiation delivered to the primary sites for over 7 weeks and concurrent chemotherapy. Oral microbiota changes in three patients were analyzed by next-generation sequencing using 16S rRNA 454 pyrosequencing. On an average, 15,000 partial 16S rRNA gene sequences were obtained from each sample. All sequences fell into 11 different bacterial phyla. During early CCRT, the microbial diversity gradually decreased. In a patient, who did not receive any antibiotics during the CCRT, Firmicutes and Proteobacteria were the most abundant phylum. During the early CCRT, proteobacteria gradually decreased while Firmicutes increased. During the late CCRT, firmicutes gradually decreased while Bacteroides and Fusobacteria increased. In all the patients, yellow complex showed a gradual decrease, while orange and red complex showed a gradual increase during the CCRT. At 4 weeks after CCRT, the recovery of oral microbiota diversity was limited. During CCRT, there was a gradual increase in major periodontopathogens in association with the deterioration of the oral hygiene. Henceforth, it is proposed that understanding oral microbiota shift should provide better information for the development of effective oral care programs for patients receiving CCRT for HNSCC.

Abnormal HLA-G expression occurs in various diseases such as melanoma, renal cell carcinoma, asthma, and classic Hodgkin’s lymphoma. The purpose of this study was to determine whether HLA-G gene is linked with oral squamous cell carcinoma (OSCC). To investigate the possible link with susceptibility to OSCC, 54 OSCC patients and 120 healthy controls were enrolled in this study. HLA-G 14bp insertion/deletion polymorphism is in 3'-untranslated region of HLA-G gene. HLA-G 14bp insertion/deletion polymorphism was analyzed using the polymerase chain reaction (PCR) method. For the analysis of genetic data, SPSS18.0 program was used. Logistic regression models were performed for odds ratio (OR), 95 percent confidence interval (CI), and p value. There was a significant difference in distribution allele between OSCC patients and control subjects (OR=0.018, 95% CI=0.002- 0.131, p<0.001). Our results suggest that HLA-G 14bp insertion/deletion polymorphism may be linked with susceptibility to OSCC in the Korean population.

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.

Anthricin (Deoxypodophyllotoxin), a naturally occurring flavolignan, has well known anti-cancer properties in several cancer cells, such as prostate cancer, cervical carcinoma and pancreatic cancer. However, the effects of Anthricin are currently unknown in oral cancer. We examined the anticancer effect and mechanism of action of Anthricin in human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that Anthricin inhibits cell viability in a dose- and time-dependent manner (IC50 50 nM) in the MTT assay and Live & Dead assay. In addition, Anthricin treated FaDu cells showed marked apoptosis by DAPI stain and FACS. Furthermore, Anthricin activates anti-apoptotic factors such as caspase-3, -9 and poly (ADP-ribose) polymerase (PARP), suggesting that caspase-mediated pathways are involved in Anthricin- induced apoptosis. Anthricin treatment also leads to accumulation of the pro-apoptotic factor Bax, followed by inhibition of cell growth. Taken together, these results indicate that Anthricn-induced cell death of human FaDu hypopharyngeal squamous carcinoma cells is mediated by mitochondrial-dependent apoptotic pathway. In summary, our findings provide a framework for further exploration on Anthricin as a novel chemotherapeutic drug for 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.

cording to histopathological characteristics. One is composite tumor, and the other is collision tumor. Composite tumor is a lesion showing the combined histopathological characteristics of two or more previously recognized tumor and/or cysts of different categories. Two constituent parts are mixed with each other, and histological transition is often observed. On the other hand, collision tumor is a lesion showing the independent histopathological characteristics of two or more previously recognized tumor and/or cysts of different categories. Two constituent parts are separated each other by connective tissue, and histological transition is not observed. Their origin is still unclear. In this study, a collision tumor was reported. 77-year old woman was referred to CNUH with a neck mass in right submandibular area. Incisional biopsy was carried out, and the patient was diagnosed as collision tumor of squamous cell carcinoma and neurofibroma. After reviewing studies reporting collision tumor from head and neck area, formation of a collision tumor was most frequent from a cervical area and average age is 59.6. A collision tumor commonly lead to misdiagnosis, and the prognosis is not always transparent. The variety symptom of collision tumor has to be kept in mind in the diagnosis and treatment of the lesions in head and neck area.

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.

소장의 종양은 매우 드물게 발생하며 그중 선암의 빈도가 가장 많고, 그 외에 악성흑색종, 유암종, 림프종, 육종 등이 주로 확인되었다. 원발부위가 자궁경부인 소장의 편평세포 암 전이는 소장종양 중에서도 매우 드물며 자궁경부암의 소장 단독전이는 현재까지 한국에서 1예가 보고되었다. 본 저자들은 매우 드문 증례인 자궁경부암의 십이지장 전이와 이에 대한 고찰을 보고하는 바이다.

Shikonin, a major ingredient in the traditional Chinese herb Lithospermumerythrorhizon, exhibits multiple biological functions including antimicrobial, anti-inflammatory, and antitumor effects. It has recently been reported that shikonin displays antitumor properties in many cancers. This study was aimed to investigate whether shikonin could inhibit oral squamous carcinoma cell (OSCC) growth via mechanisms of apoptosis and cell cycle arrest. The effects of shikonin on the viability and growth of OSCC cell line, SCC25 cells were assessed by MTT assay and clonogenic assays, respectively. Hoechst staining and DNA electrophoresis indicated that the shikonin-treated SCC25 cells were undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, flow cytometry, MMP activity, and proteasome activity also supported the finding that shikonin induces apoptosis. Shikonin treatment of SCC25 cells resulted in a time- and dose-dependent decrease in cell viability, inhibition of cell growth, and increase in apoptotic cell death. The treated SCC25 cells showed several lines of apoptotic manifestation as follows: nuclear condensation; DNA fragmentation; reduced MMP and proteasome activity; decrease in DNA contents; release of cytochrome c into cytosol; translocation of AIF and DFF40 (CAD) onto the nuclei; a significant shift in Bax/Bcl-2 ratio; and activation of caspase-9, -7, -6, and -3, as well as PARP, lamin A/C, and DFF45 (ICAD). Shikonin treatment also resulted in down-regulation of the G1 cell cycle-related proteins and up-regulation of p27KIP1. Taken together, our present findings demonstrate that shikonin strongly inhibits cell proliferation by modulating the expression of the G1 cell cycle-related proteins, and that it induces apoptosis via the proteasome, mitochondria, and caspase cascades in SCC25 cells.

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.

Fluoride has been accepted as an important material for oral health and is widely used to prevent dental caries in dentistry. However, its safety is still questioned by some. Autophagy has been implicated in cancer cell survival and death, and may play an important role in oral cancer. This study was undertaken to examine whether sodium fluoride (NaF) modulates autophagy in SCC25 human tongue squamous cell carcinoma cells. NaF demonstrated anticancer activity via autophagic and apoptotic cell death. Autophagic vacuoles were detectable using observed to form by monodansylcadaverine (MDC) and acridine orange (AO). Analysis of NaF-treated SCC25 cells for the presence of biochemical markers revealed direct effects on the conversion of LC-3II, degradation of p62/SQSTM1, cleavage formation of ATG5 and Beclin-1, and caspase activation. NaF-induced cell death was suppressed by the autophagy inhibitor 3-methyladenine (3-MA). NaF-induced autophagy was confirmed as a pro-death signal in SCC25 cells. These results implicate NaF as a novel anticancer compound for oral cancer therapy.

A poor prognosis of oral squamous cell carcinoma (SCC) is partly due to the invasiveness and metastasis of the tumor. One of key elements in tumor invasion and metastasis in the degradation of extracellular matrix is tissue inhibitors of metalloproteinases (TIMPs). This study was performed to determine the expression of TIMP-1 and TIMP-2 of oral SCCs with regard to the histologic invasiveness and differentiation in 5 normal oral mucosa and 36 oral SCCs. The histologic invasiveness of oral SCCs were classified into 4 grades. The differentiation of oral SCCs was divided into 3 grades. The StreptAvidin-Biotin immunohistochemical process, using TIMP-1 and TIMP-2 monoclonal antibodies, was performed to determine the expression of TIMP-1 and TIMP-2. The expression of TIMP-1 was positive in 5 of 17 oral SCCs with weak invasiveness and was positive in 8 of 19 oral SCCs with strong invasiveness. The TIMP-1 expression did not increase significantly with respect to the invasiveness of oral SCCs (P>0.05). The expression of TIMP-2 was strongly positive in 5 out of 17 SCCs with weak invasiveness and was strongly positive in 15 of 19 SCCs with strong invasiveness. The TIMP-2 expression increased significantly with respect to the invasiveness of oral SCCs; the stronger the expression, the stronger the invasiveness (P<0.05). The expression of TIMP-1 and TIMP-2 did not increase significantly with respect to the histologic differentiation. We concluded that with respect to the invasiveness, the TIMP-2 expression increases significantly in oral SCCs but the TIMP-1 expression does not; and that with respect to the histologic differentiation, their expressions do not increase significantly. These results suggested that TIMP-2 can be used as a tool to evaluate the invasiveness of oral SCCs.

Curcumin is a widely used flavoring agent in food, and it has been reported to inhibit cell growth, to induce apoptosis, and to have antitumor activity in many cancers. Cisplatin is one of the most potent known anticancer agents and shows significant clinical activity against a variety of solid tumors. This study was undertaken to investigate the synergistic apoptotic effects of co-treatment with curcumin and cisplatin on human tongue SCC25 cells. To investigate whether the co-treatment efficiently reduced the viability of the SCC25 cells compared with the two treatments separately, an MTT assay was conducted. The induction and the augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and an analysis of DNA hypoploidy. Western blot, MMP and immunofluorescence tests were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following the co-treatment. In this study, following the co-treatment with curcumin and cisplatin, the SCC25 cells showed several forms of apoptotic manifestation, such as nuclear condensation, DNA fragmentation, reduction of MMP, increased levels of Bax, decreased levels of Bcl-2, and decreased DNA content. In addition, they showed a release of cytochrome c into the cytosol, translocation of AIF and DFF40 (CAD) to the nuclei, and activation of caspase-7, caspase-3, PARP, and DFF45 (ICAD). In contrast, separate treatments of 5 μM of curcumin or 4 μg/ml of cisplatin, for 24 hours, did not induce apoptosis. Therefore, our data suggest that combination therapy with curcumin and cisplatin could be considered as a novel therapeutic strategy for human oral squamous cell carcinoma.

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.