鵜澤 一弘

Cetuximab, an inhibitor of the epidermal growth factor receptor that is used widely to treat human cancers including oral squamous cell carcinoma (OSCC), has characteristic side effects of skin rash and hypomagnesemia. However, the mechanisms of and therapeutic agents for skin rashes and hypomagnesemia are still poorly understood. Our gene expression profiling analyses showed that cetuximab activates the p38 MAPK pathways in human skin cells (human keratinocyte cell line [HaCaT]) and inhibits c-Fos-related signals in human embryonic kidney cells (HEK293). We found that while the p38 inhibitor SB203580 inhibited the expression of p38 MAPK targets in HaCaT cells, flavagline reactivated c-Fos-related factors in HEK293 cells. It is noteworthy that, in addition to not interfering with the effect of cetuximab by both compounds, flavagline has additive effect for OSCC growth inhibition in vivo. Collectively, our results indicate that combination of cetuximab and these potential therapeutic agents for cetuximab-related toxicities could be a promising therapeutic strategy for patients with OSCC.

Tripartite motif family-like 2 (TRIML2), a member of the TRIM proteins family, is closely related to Alzheimer's disease, however, no studies of TRIML2 have been published in the cancer research literature. In the current study, we investigated the expression level of TRIML2 and its molecular mechanisms in human oral squamous cell carcinoma (OSCC); reverse transcriptase-quantitative polymerase chain reaction, immunoblot analysis, and immunohistochemistry showed that TRIML2 is up-regulated significantly in OSCCs in vitro and in vivo. TRIML2 knockdown OSCC cells showed decreased cellular proliferation by cell-cycle arrest at G1 phase that resulted from down-regulation of CDK4, CDK6, and cyclin D1 and up-regulation of p21Cip1 and p27Kip1. Surprisingly, resveratrol, a polyphenol, led to not only down-regulation of TRIML2 but also cell-cycle arrest at G1 phase similar to TRIML2 knockdown experiments. Taken together, we concluded that TRIML2 might play a significant role in tumoral growth and that resveratrol may be a new drug for treating OSCC by interfering with TRIML2 function.

Synaptotagmin12 (SYT12) has been well characterized as the regulator of transmitter release in the nervous system, however the relevance and molecular mechanisms of SYT12 in oral squamous cell carcinoma (OSCC) are not understood. In the current study, we investigated the expression of SYT12 and its molecular biological functions in OSCC by quantitative reverse transcriptase polymerase chain reaction, immunoblot analysis, and immunohistochemistry. SYT12 were up-regulated significantly in OSCC-derived cell lines and primary OSCC tissue compared with the normal counterparts (P<0.05) and the SYT12 expression levels were correlated significantly with clinical indicators, such as the primary tumoral size, lymph node metastasis, and TNM stage (P<0.05). SYT12 knockdown OSCC cells showed depressed cellular proliferation, migration, and invasion with cell cycle arrest at G1 phase. Surprisingly, we found increased calcium/calmodulin-dependent protein kinase 2 (CAMK2) inhibitor 1 (CAMK2N1) and decreased CAMK2-phosphorylation in the knockdown cells. Furthermore, treatment with L-3, 4-dihydroxyphenylalanine (L-dopa), a drug approved for Parkinson's disease, led to down-regulation of SYT12 and similar phenotypes to SYT12 knockdown cells. Taken together, we concluded that SYT12 plays a significant role in OSCC progression via CAMK2N1 and CAMK2, and that L-dopa would be a new drug for OSCC treatment through the SYT12 expression.

Filamin-binding LIM protein 1 (FBLIM1) is related to regulation of inflammatory responses, such as chronic recurrent multifocal osteomyelitis; however, the relevance of FBLIM1 in oral squamous cell carcinoma (OSCC) is unknown. The aim of the current study was to elucidate the possible role of FBLIM1 in the carcinogenesis of OSCC. We analyzed FBLIM1 expression using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR), immunoblot analysis, and immunohistochemistry. The expression levels of FBLIM1 were up-regulated significantly (P < 0.05) in OSCC-derived cell lines and primary OSCCs specimens compared with normal counterparts. FBLIM1 expression also was correlated with the primary tumoral size (P < 0.05) and vascular invasion (P < 0.05). We then assessed tumoral progression after treatment with FBLIM1 siRNA and clopidogrel, an antiplatelet agent. Similar to the FBLIM1 knockdown effect, clopidogrel-treated cells had attenuated functions of proliferation, migration, and invasiveness. Interestingly, clopidogrel treatment led to down-regulation of epidermal growth factor receptor (EGFR) and FBLIM1. These findings identify FBLIM1 as a putative therapeutic target by using clopidogrel for inhibiting over activation of EGFR signaling to prevent OSCC malignancy.

症例は73歳女性で、左側上顎歯肉の違和感を主訴に受診した。パノラマX線写真で左上7番の根尖付近から上顎骨および上顎洞部にかけてのX線不透過像を指摘された。パノラマX線写真にて左上7番の根尖付近から上顎骨内に存在し、先端が上顎洞底線を越えて上顎洞内へ至るX線不透過像を認めた。デンタルX線写真では根分岐部から異物の断端にかけてX線透過像を認めた。CT写真では口蓋根根尖から上顎洞内にかけて約15mmの金属針様異物を認めた。左上7番は画像所見、臨床症状より保存不可能と判断し、局所麻酔下にて左上7番抜歯術および異物除去術を行った。抜歯したところ、根分岐部から口蓋根尖にかけて肉芽組織を認めた。抜歯窩の肉芽組織を掻爬し、抜歯窩骨面を精査すると、異物の断端を確認した。術後の抜歯窩の治癒経過は良好で、術前に訴えていた左側上顎歯肉の違和感は消失した。術後2ヵ月の診察時に口腔上顎洞瘻孔や副鼻腔炎症状を認めないことを確認し、経過観察を終了とした。

口腔外傷を主訴に受診した精神遅滞患者47例(男性28例、女性19例、平均31.6±15.3歳)を対象とした。既往歴は、てんかん27例、自閉症13例、Down症候群2例、脳性麻痺および統合失調症各1例であった。受傷契機は転倒が27例で最も多く、自分自身に危害を加えて受傷した自傷18例、他者から危害を受けて受傷した他害2例の順であった。受傷時期は3月、6月、12月に多く、季節では3月から5月の春が13例、6月から8月の夏が14例、9月から11月の秋が7例、12月から2月の冬が13例であった。外傷の種別では、歯の単独損傷が31例と最も多く、次いで軟組織単独損傷8例、歯牙と軟組織の合併損傷が7例、軟組織と顎骨の合併損傷が1例の順であった。歯の損傷に対しては歯冠修復抜髄および暫間固定などの保存的処置が17例、抜歯を行った外科処置が16例、経過観察が5例で、約半数が外科処置を選択した。軟組織損傷15例については縫合を行ったものが9例、経過観察が6例であった。顎骨の損傷の1例については整復不可能であったため、小骨片除去が行われた。

Multiple coagulation factor deficiency protein 2 (MCFD2), a binding partner of lectin mannose binding 1 (LMAN1), causes combined deficiencies of coagulation factors V and VIII. MCFD2 function in inherited hematologic disorders is well elucidated however, little is known about its role in human tumorigenesis. The aim of the current study was to investigate the states of MCFD2 in oral squamous cell carcinoma (OSCC). The expression of MCFD2 was up-regulated significantly in all cell lines examined. Evaluation of the cellular functions associated with tumoral metastasis showed that MCFD2 knockdown (shMCFD2) cells exhibited significantly lower cellular invasiveness and migration and higher cellular adhesion compared with shControl cells. Of note, shMCFD2 cells also showed weak immunoreactivity of LMAN1 and a lower secretion level of galactoside-binding soluble 3 binding protein (LGALS3BP). In addition to in vitro validation, clinical data on 70 patients with OSCC indicated that state of MCFD2 expression level is associated with regional lymph node metastasis. Altogether, we have demonstrated that MCFD2 promotes cancer metastasis by regulating LMAN1 and LGALS3BP expression levels. Hence, MCFD2 may represent a promising candidate for a novel therapeutic target for patients with metastatic OSCCs.

Schwannomas are typically solitary benign neural tumors however, multiple lesions associated with the rare genetic disorder neurofibromatosis type 2 (NF2) have been reported in some cases. We present the case of a tongue schwannomas in a 36-year-old woman previously diagnosed with NF2 with bilateral vestibular schwannomas. To alleviate difficulties with swallowing, tongue nodular masses were surgically removed, and schwannoma was histologically diagnosed. Our patient represents the first case of histologically confirmed tongue schwannoma associated with NF2. This indicates that in patient with NF2, schwannoma may be detected in the oral cavity as well as in other parts of the body. Thus, careful clinical and histological examinations are warranted to identify schwannomas associated with NF2 even in the oral cavity.

Diacylglycerol lipase alpha (DAGLA), which catalyzes the hydrolysis of diacylglycerol to 2-arachidonoylglycerol and free fatty acid, is required for axonal growth during the brain development and for retrograde synaptic signaling at mature synapses. So far, no information was found regarding the possible role of DAGLA in human tumorigenesis. Thus, the current study sought to clarify the contribution of DAGLA in oral squamous cell carcinomas (OSCCs) and assess the clinical possibilities for OSCC treatment. Using real-time quantitative reverse transcription-polymerase chain reaction, immunoblotting, and immunohistochemistry, we found a significant up-regulation of DAGLA in OSCCs compared with normal cells and tissues both at mRNA and protein expression levels. Knockdown models in OSCC-derived cell lines for DAGLA (siDAGLA) and treatment with a lipase inhibitor (orlistat) showed several depressed cellular functions, including cellular proliferation and migratory activities through cell-cycle arrest at G1 phase. Furthermore, we found that DAGLA-positive OSCC samples were correlated highly with the primary tumoral size. We concluded that DAGLA may be a key determinant in tumoral progression and might be a therapeutic target for OSCCs.

Deoxynucleotidyl transferase terminal interacting protein 1 (DNTTIP1) forms a complex with histone deacetylase (HDAC) however, the relevance of DNTTIP1 in cancer remains unknown. The aim of this study was to examine DNTTIP1 expression and its functional mechanisms in oral squamous cell carcinomas (OSCCs). DNTTIP1 expression was analyzed by quantitative reverse transcriptase-polymerase chain reaction, immunoblotting analysis, and immunohistochemistry. The expression of DNTTIP1 was upregulated significantly in vitro and in vivo, and in patients with OSCC in whom DNTTIP1 was overexpressed and the expression level was correlated significantly (P &lt 0.05) with tumoral growth. DNTTIP1 knockdown (siDNTTIP1) cells showed depressed cellular proliferation by cell-cycle arrest at the G1 phase with high acetylation of p53 and upregulation of p21Cip1. Moreover, resveratrol, a HDAC inhibitor, controlled not only acetylated p53 status but also DNTTIP1 expression, leading to a similar phenotype of siDNTTIP1 cells. A marked (P &lt 0.05) reduction of tumoral growth in mouse xenograft models was observed with lower DNTTIP1 expression under the presence of this chemical reagent. Taken together, our results suggested that DNTTIP1–HDAC interaction promotes tumoral growth through deacetylation of p53 and that DNTTIP1 might be a critical therapeutic target in OSCCs.

Angiopoietin-1 (Ang1) is a binding partner of endothelial cell-specific tyrosine-protein kinase receptor (Tie2), which serves important roles in vascular development and angiogenesis. Tie2 is closely associated with the metastasis of oral squamous cell carcinomas (OSCCs) however, little is known about the correlation between Tie2 and Ang1. In the present study, the functional mechanisms of the Tie2/Ang1 interaction were investigated using Tie2 overexpressed (oeTie2) OSCC cells and recombinant Ang1 protein. oeTie2 cells had increased cell-cell and cell-extracellular matrix adhesions compared with the control cells. Additionally, the adhesive activities increased following treatment with exogenous Ang1, indicating that Ang1 directly enhances Tie2 functions. In the clinical OSCC data from 10 cases positive for regional lymph node metastasis, all cases were negative for Tie2 expression and eight cases (80%) were negative for Ang1 expression. These results suggest that Tie2 and Ang1 serve important roles in cancer metastasis and may be potential biomarkers and therapeutic targets for OSCC metastasis.

症例は91歳女性で、歩行中に転倒してオトガイ部を強打し受傷した。両側下顎骨骨折の診断を受けた。パノラマX線写真およびCT写真にて右側下顎骨骨体部(第一・第二小臼歯相当部)に骨折線、左側下顎骨骨体部(中切歯から第一大臼歯相当部にかけて)に頬舌側的に骨折線を認めた。また、両側下顎骨骨折によるオトガイ棘の後方偏位およびそれに伴った舌根沈下と上気道狭窄を認めた。呼吸不全やSpO2の低下は認めなかったが、画像検査により舌根沈下および上気道狭窄を疑い、呼吸管理目的に入院とした。第10病日に全身麻酔下において両側下顎骨骨折観血的整復固定術を施行した。気道確保は経鼻挿管で行った。手術は口内法で行った。創部への感染は認めず、術後第9病日で軽快退院した。術後パノラマX線写真にて下顎骨の整復、CT写真で下顎骨整復による舌根沈下および上気道狭窄が改善していることを確認した。術後4ヵ月に下顎義歯を新製した。術後18ヵ月経過した現在、経過良好である。

Tryptophan-aspartic acid (WD) repeat-containing protein 34 (WDR34), one of the WDR protein superfamilies with five WD40 domains, inhibits a transforming growth factor-beta (TGF-β) activated kinase 1 (TAK1)-associated NF-κB activation pathway. Nevertheless, little is known about the roles of WDR34 in cancer. The current study sought to elucidate the clinical relevance of WDRsfb34 in oral squamous cell carcinoma (OSCC). We found WDR34 down-regulation in OSCCs compared with normal control tissues using real-time quantitative reverse transcription-polymerase chain reaction, immunoblotting, and immunohistochemistry. Models of overexpression of WDR34 (oeWDR34) showed depressed cellular growth through cell-cycle arrest at the G1 phase. To investigate the inhibitory function of WDR34, we challenged oeWDR34 cells with interleukin (IL)-1, a ligand for activation of the TAK1-NF-κB pathway and assessed the expression of a target gene of the pathway. oeWDR34 strongly inhibited IL-6 expression, which is closely related to tumoral growth, compared with control cells, suggesting that WDR34 would be a critical molecule for control of tumoral progression. In addition to the in vitro experiments, WDR34 negativity was correlated with tumoral growth of OSCCs. Our findings suggested that WDR34 inhibits OSCC progression and might be a potential tumor-suppressor molecule in OSCCs.

症例は24歳女性で、左側頬部に腫脹を認め、腫脹は増大・縮小を繰り返した。左側頬粘膜下に直径約10mm大、弾性軟、表面滑沢な腫瘤を口腔内外より触知した。CT所見では、左側頬粘膜下に直径約10mm大の下顎骨との連続性がない腫瘤を認めた。周囲組織への浸潤は認めなかった。左側頬粘膜下良性腫瘍と診断し、全身麻酔下にて病変切除術を施行した。病変は一部周囲組織との癒着を認めたため、周囲組織を含めて切除した。術後1年経過した現在、再発は認めず、経過良好である。病理組織学的所見により結節性筋膜炎と診断した。

Mucoepidermoid carcinoma (MEC) is the most common malignant neoplasm of the salivary gland. Albeit common, histologic variants have rarely been noted in MEC. Here, we report a 49-year-old man with a sublingual gland tumor. Histologically, the tumor was composed of spindle cells arranged in interlacing fascicules or globular nests. A few bland small glands containing mucous cells were also scattered. The spindle tumor cells completely lacked immunoreactivity for cytokeratin, and exhibited immunoreactivity for vimentin, S-100, HMB-45, Melan A, and SOX10. The tumor was initially suspected to be clear cell sarcoma, malignant melanoma, or perivascular epithelioid cell tumor with a few entrapped nonneoplastic duct epitheliums. However, reverse-transcription polymerase chain reaction revealed the CRTC3-MAMI2 fusion gene product diagnostic of MEC. In fact, a very minor component of the epithelial cells was reminiscent of conventional MEC, whereas major spindled tumor cells possessed markedly altered differentiation. This is the first case report of MEC with extensive spindled morphology and melanocytic marker expression. (C) 2017 Elsevier Inc. All rights reserved.

Dentinogenic ghost cell tumors (DGCTs) are extremely rare, accounting for less than 0.5% of all odontogenic tumors. DGCTs were described initially as solid neoplasms of calcifying odontogenic cysts (COCs). In the 2005 WHO Histological Classification of Odontogenic Tumors, COCs were classified as tumors and renamed as calcifying cystic odontogenic tumors (CCOTs). The term of DGCT was retained and histologically defined as a locally invasive neoplasm characterized by ameloblastoma-like islands of epithelial cells in a mature connective tissue stroma, aberrant keratinization comprised of ghost cells, and an association with dysplastic dentin. DGCTs were subdivided into two variants: intraosseous (central) and extraosseous (peripheral). Generally, peripheral DGCTs occur less frequently than central ones. We report a rare case of a peripheral DGCT in a 60-year-old man with a well-circumscribed nodule in the left canine region of the maxilla. The lesion measured about 11 millimeters in diameter. No bone involvement was observed radiographically. Based on clinical and radiographic findings, the tumor was provisional diagnosed as benign. Consequently, it was enucleated completely, and a histopathological diagnosis of DGCT was made. The patient has remained disease-free for 18 months postoperatively. The biologic behavior of central DCGTs is considered to be more aggressive and require radical treatment. In contrast, peripheral DGCTs have been reported to be non-aggressive, because local bone resorption or recurrences have not yet been reported. There are no differences in the histopathologic features between the central and peripheral variants. Accordingly, accurate preoperative evaluation is crucial to differentiate between these lesions including other odontogenic tumors.

歯周嚢胞は歯の歯頸部付近にみられる炎症性の歯原性嚢胞であり,歯周ポケットの炎症過程を経て発生する。上皮性嚢胞が好発する顎骨において稀な疾患で,中でも第三大臼歯以外に発生する例は報告が少ない。症例は12歳男性。近歯科を受診した際に,X線検査にて嚢胞様病変を指摘され当科を紹介され受診した。CT画像上,左側下顎第二大臼歯の頬側に接する単房性,類円形の透過像を認めた。全身麻酔下に嚢胞摘出術を施行した。その際,第二大臼歯は温存した。病理組織学的検査にて高度の炎症性細胞浸潤を伴う嚢胞壁を認めた。臨床所見,X線検査所見および病理組織学的所見から歯周嚢胞と最終診断した。術後1年間経過観察を行ったが,再発所見は認めず経過良好である。(著者抄録)

Ubiquitin-conjugating enzyme E2S (UBE2S), a family of E2 protein in the ubiquitin-proteasome system, is highly expressed in several types of cancers; however, its roles in oral squamous cell carcinoma (OSCC) have not yet been well elucidated. The purpose of this study was to clarify the functional activities of UBE2S in OSCCs. We analyzed the expression levels of UBE2S in nine OSCC cell lines and primary OSCC tissues by quantitative reverse transcriptase-polymerase chain reaction, Western blotting, and immunohistochemistry (IHC). The correlations between UBE2S expression and clinical classifications of OSCCs were analyzed using the IHC scoring system. We also used UBE2S knockdown OSCC cells for functional assays (proliferation assay, flow cytometry, and Western blotting). UBE2S was overexpressed in OSCCs in vitro and in vivo and was correlated significantly (P &lt; 0.05) with the primary tumoral size. The cellular growth was decreased and the cell-cycle was arrested in the G2/M phase in the UBE2S knockdown (shUBE2S) cells. The expression level of P21, a target of the ubiquitin-proteasome system, was increased in the shUBE2S cells because of lower anaphase activity that promotes complex subunit 3 (APC3), an E3 ubiquitin ligase, compared with shMock cells. These findings might promote the understanding of the relationship between UBE2S overexpression and oral cancer proliferation, indicating that UBE2S would be a potential biomarker of and therapeutic target in OSCCs. (C) 2017 Elsevier Inc. All rights reserved.

Resistance to anticancer medications often leads to poor outcomes. The present study explored an effective approach for enhancing chemotherapy targeted against human cancer cells. Real-time quantitative real-time polymerase chain reaction (qRT-PCR) analysis revealed overexpression of members of aldo-keto reductase (AKR) 1C family, AKR1C1, AKR1C2, AKR1C3, and AKR1C4, in cisplatin, cis-diamminedichloroplatinum (II) (CDDP)resistant human cancer cell lines, HeLa (cervical cancer cells) and Sa3 (oral squamous cell carcinoma cells). The genes were downregulated using small-interfering RNA (siRNA) transfection, and the sensitivity to CDDP or 5-fluorouracil (5-FU) was investigated. When the genes were knocked down, sensitivity to CDDP and 5-FU was restored. Furthermore, we found that administration of mefenamic acid, a widely used non-steroidal anti-inflammatory drug (NSAID) and a known inhibitor of AKR1Cs, enhanced sensitivity to CDDP and 5-FU. The present study suggests that AKR1C family is closely associated with drug resistance to CDDP and 5-FU, and mefenamic acid enhances their sensitivity through its inhibitory activity in drug-resistant human cancer cells. Thus, the use of mefenamic acid to control biological function of AKR1C may lead to effective clinical outcomes by overcoming anticancer drug resistance.

TEA domain transcription factor 4 (TEAD4), which has critical functions in the process of embryonic development, is expressed in various cancers. However, the important role of TEAD4 in human oral squamous cell carcinomas (OSCCs) remain unclear. Here we investigated the TEAD4 expression level and the functional mechanism in OSCC using quantitative reverse transcriptase-polymerase chain reaction, Western blot analysis, and immunohistochemistry. Furthermore, TEAD4 knockdown model was used to evaluate cellular proliferation, cell-cycle analysis, and the interaction between TEAD4 and Yes-associated protein (YAP) which was reported to be a transcription coactivator of cellular proliferation. In the current study, we found that TEAD4 expression increased significantly in vitro and in vivo and correlated with tumoral size in OSCC patients. TEAD4 knockdown OSCC cells showed decreased cellular proliferation resulting from cell-cycle arrest in the G1 phase by down-regulation of cyclins, cyclin-dependent kinases (CDKs), and up-regulation of CDK inhibitors. We also found that the TEAD4-YAP complex in the nuclei may be related closely to transcriptions of G1 arrest-related genes. Taken together, we concluded that TEAD4 might play an important role in tumoral growth and have potential to be a therapeutic target in OSCCs. (C) 2017 Elsevier Inc. All rights reserved.

Signal-induced proliferation-associated protein 1 (SIPA1) is known to be a GTPase activating protein. Overexpressed SIPA1 is related to metastatic progression in breast and prostate cancers; however, the relevance of SIPA1 in oral squamous cell carcinoma (OSCC) is still unknown. The aim of this study was to examine SIPA1 expression and its functional mechanisms in OSCC. SIPA1 mRNA and protein expressions were analyzed by quantitative reverse transcriptase-polymerase chain reaction, Western blot analysis, and immunohistochemistry. The expressions of SIPA1 were up-regulated significantly in vitro and in vivo. Moreover, SIPA1 expression was correlated with regional lymph node metastasis. We next assessed the cellular functions associated with tumoral metastasis using SIPA1 knockdown (shSIPA1) cells and analyzed the downstream molecules of SIPA1, i.e., bromodomain containing protein 4(BRD4), integrin betal (ITGB1), and matrix metalloproteinase 7 (MMP7). The shSIPA1 cells showed decreased invasiveness and migratory activities, however cellular adhesion ability was maintained at a high level. In addition, ITGB1 expression was greater in shSIPA1 cells, whereas MMP7 expression was lower than in control cells. This research is the first to establish that SIPA1 promotes cancer metastasis by regulating the ITGB1 and MMP7. Therefore, SIPA1 might be a novel therapeutic target for patients with lymph node metastasis of OSCC.

Lymphocyte cytosolic protein 1 (LCP1), a member of actin-binding protein of the plastin family, has been identified in several malignant tumors of non-hematopoietic sites, such as the colon, prostate, and breast. However, little is known about the roles of LCP1 in oral squamous cell carcinomas (OSCCs). This present study sought to clarify the clinical relevance of LCP1 in OSCCs and investigate possible clinical applications for treating OSCCs by regulating LCP1 expression. We found up-regulation of LCP1in OSCCs compared with normal counterparts using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunoblotting, and immunohistochemistry (P &lt; 0.05). We used shRNA models for LCP1 (shLCP1) and enoxacin (ENX), a fluoroquinolone antibiotic drug, as a regulator of LCP1 expression. In addition to the LCP1 knockdown experiments in which shLCP1 cells showed several depressed functions, including cellular proliferation, invasiveness, and migratory activities, ENX-treated cells also had attenuated functions. Consistent with our hypothesis from our in vitro data, LCP1-positive OSCC samples were correlated closely with the primary tumoral size and regional lymph node metastasis. These results suggested that LCP1 is a useful biomarker for determining progression of OSCCs and that ENX might be a new therapeutic agent for treating OSCCs by controlling LCP1 expression.

Zrt-Irt-like protein 4 (ZIP4) is critical molecule for proper mammalian development and releasing zinc from vesicular compartments. Recent studies suggested that ZIP4 plays an important role of tumor progression in pancreatic, prostate, and hepatocellular cancers, however, little is known about the detail mechanism of ZIP4 in their cancers. In the present study, we examined the possibility of ZIP4 as a new molecular target for oral squamous cell carcinoma (OSCC). We evaluated ZIP4 expression in OSCC-derived cell lines and primary OSCC samples by quantitative RT-PCR, immunoblotting, and immunohistochemistry (IHC). We also analyzed the clinical correlation between ZIP4 status and clinical behaviors in patients with OSCC. In addition, ZIP4 knockdown cells (shZIP4 cells) and ZnCl2 treatment were used for functional experiments, including cellular proliferation assay, zinc uptake assay, and cell-cycle analysis. ZIP4 mRNA and protein were up-regulated significantly in OSCCs compared with normal counterparts in vitro and in vivo. IHC showed that ZIP4 expression in the primary OSCC was positively correlated with primary tumoral size. The shZIP4 cells showed decrease accumulation of intercellular zinc and decreased cellular growth by cell-cycle arrest at the G1 phase, resulting from up-regulation of cyclin-dependent kinase inhibitors and down-regulation of cyclins and cyclin-dependent kinases. Since cellular growth of OSCC cells after treatment with zinc was significantly greater than control cells, we speculated that intercellular ZnCl2 accumulation is an important factor for cellular growth. Consistent with our hypothesis, not only decreased zinc uptake by ZIP4 knockdown but also chelating agent, N, N, N', N'-tetrakis(2-pyridylmethyl) ethylenediamine (TPEN), showed inhibitory effects of cellular proliferation. Therefore, our data provide evidence for an essential role of ZIP4 and intracellular zinc for tumoral growth in OSCC, suggesting that zinc uptake might be a potential therapeutic targeting event for OSCCs. (C) 2016 Elsevier Inc. All rights reserved.

Activin B, a homodimer of inhibin beta b (INHBB), is a multifunctional cytokine belonging to the transforming growth factor-beta (TGF-beta) family. However, the molecular functions and clinical relevance of activin B have not been determined in oral cancer. We investigated the critical roles of activin B in oral squamous cell carcinoma (OSCC). We performed quantitative reverse transcriptase-polymerase chain reaction, Western blotting, and immunohistochemistry to study INHBB expression in OSCC-derived cell lines and OSCC clinical samples. The INHBB expression levels were significantly (P &lt; 0.05) overexpressed in OSCCs compared to normal counterparts in vitro and in vivo. Activin B-positivity in OSCC cases was significantly (P &lt; 0.05) correlated with regional lymph node metastasis. The INHBB knockdown (shINHBB) cells promoted cellular adhesion and suppression of cellular invasiveness and migration. After treatment of shINHBB cells with activin B, those activities were restored similar to the shMock cells. In the processes of invasiveness and metastasis, the cells cause epithelial-mesenchymal transition (EMT). TGF-beta and its family members are promoters of the EMT process. To investigate whether activin B is related to EMT, we examined the expressions of EMT-related genes and found that INHBB was related closely to EMT. Our results suggested for the first time that activin B indicates tumoral metastasis in OSCCs and might be a useful biomarker for OSCC metastasis.

Because of cellular senescence/apoptosis, no effective culture systems are available to maintain replication of cells from odontogenic tumors especially for odontoma, and, thus, the ability to isolate human odontoma-derived cells (hODCs) for functional studies is needed. The current study was undertaken to develop an approach to isolate hODCs and fully characterize the cells in vitro. The hODCs were cultured successfully with a Rho-associated protein kinase inhibitor (Y-27632) for an extended period with stabilized lengths of the telomeres to sustain a similar phenotype/property as the primary tumoral cells. While the hODCs showed stable long-term expansion with expression of major dental epithelial markers including dentin sialophosphoprotein (DSPP) even in the three-dimensional microenvironment, they lack the specific markers for the characteristics of stem cells. Moreover, cells from dental pulp showed significant up-regulation of DSPP when co-cultured with the hODCs, while control fibroblasts with the hODCs did not. Taken together, we propose that the hODCs can be isolated and expanded over the long term with Y-27632 to investigate not only the development of the hODCs but also other types of benign human tumors.

Oral pigmentation ranges from physiologic pigmentation to malignant neoplasms. Blue nevi (BN), benign melanocytic lesions that rarely develop in the oral cavity, can be associated with malignant melanoma. Although most oral pigmented lesions are benign, long-term follow-up/histopathologic evaluation eliminate oral malignant melanoma (OMM) when lesions develop in the most frequent sites (palate and maxillary gingiva). A 58-year-old Japanese man with a BN presented with a 5-year history of a blue-black macule on the hard palate. The patient had no malignant signs, no abnormal radiologic findings in the maxillofacial region, or abnormal laboratory values. Histopathology showed parallel spindle-shaped melanocytes under the epithelium with fibrosis. A BN was diagnosed. The 2-year follow-up and clinical outcome were satisfactory, without recurrence or complications. Close monitoring and long-term follow-up or early biopsy to exclude OMM in the oral cavity are important in cases in which the diagnosis is not definitive clinically.

IgA血管炎は、以前Henoch-Schoenlein紫斑病と呼ばれており、IgA抗体優位の免疫複合体が小血管へ沈着することで発症し、進行するとネフローゼ症候群や腎障害を併発する疾患である。今回われわれは、84歳女性で左側上顎歯肉癌の術後33日目に両側下肢に紫斑を認めた症例を報告する。臨床症状および生検結果からIgA血管炎と診断された。ジアフェニルスルホンおよびプレドニゾロンの内服により紫斑および腎機能が改善し、以後、原疾患の上顎歯肉癌の再発および紫斑の再燃を認めず経過良好である。(著者抄録)

急性汎発性発疹性膿疱症(AGEP)は重症薬疹の1つであり、出血を伴わない膿疱性皮疹や高熱を認めることが特徴である。患者は29歳女性でインフルエンザBに対し抗インフルエンザ薬を内服後、全身および口腔内に高熱を伴う皮疹が出現した。病歴、臨床所見、血液検査の結果から感染所見がないため、AGEPと診断された。症状は内服薬の中止および皮疹部と口腔内発疹部にステロイド軟膏を塗布し改善した。退院後も再発等なく経過している。(著者抄録)

Cavin-2 (CVN2) affects formation of large caveolae, which are membrane-rich cholesterol domains associated with several functions in signal transduction. Accumulating evidence suggests that CVN2 is present in many cellular types; however, the molecular mechanisms of CVN2 in cancers and its clinical relevance are unknown. We proposed a mechanism by which CVN2 regulates caveolin-1 expression leading to slow cellular proliferation by inactivation of the extracellular regulated kinase (ERK) pathway. Quantitative reverse transcriptase-polymerase chain reaction and immunoblot analyses were used to assess the CVN2 regulation mechanism in oral squamous cell carcinoma (OSCC). Immunohistochemistry (IHC) was performed to analyze the correlation between CVN2 expression and clinical behavior in 115 patients with OSCC. A CVN2 overexpressed model of OSCC cells (oeCVN2 cells) was used for functional experiments. CVN2 expression was down-regulated significantly (P&lt;0.05) in OSCCs compared with normal counterparts in vitro and in vivo. In addition to the findings that a serum deprivation culture induced up-regulation of CVN2 and slowed cellular proliferation, oeCVN2 cell growth decreased because of cell-cycle arrest at the G1 phase resulting from up-regulated cyclin-dependent kinase inhibitors (p21(Cip1) and p27(Kip1)) and down-regulated cyclins (cyclin D1, cyclin E) and cyclin-dependent kinases (CDK2, CDK4, and CDK6). Interestingly, CVN2 overexpression facilitated caveolin-1 recruitment and colocalization with each other. We also found decreased ERK phosphorylation levels, an upstream event in cell-cycle arrest. Clinically, IHC data from primary OSCCs showed high tumoral progression in CVN2-negative patients with OSCC. CVN2 may be a possible key regulator of OSCC progression via the CVN2/caveolin-1/ERK pathway and a potential therapeutic target for developing new treatments for OSCCs. (c) 2015 Wiley Periodicals, Inc.

症例は39歳男性で、中国済南市で自動車による交通事故に遭い緊急搬送された。精密検査で下顎骨骨折、肋骨骨折、肺挫傷および出血性腸炎の診断を受けた。しかし、下顎骨骨折の治療が困難のため紹介受診した。全身に皮下出血斑を認めた。口腔外所見は右側オトガイ部皮膚に挫創を認めた。右側下唇からオトガイ部にかけ、知覚鈍麻を認めた。口腔内所見で左側下顎側切歯の歯冠および左側下顎第一大臼歯の舌側の歯冠破折を認めた。パノラマX線で右側下顎骨体部と左側関節突起部の骨折を認めた。CTで右側肋骨骨折、肺挫傷、右気管支に気道異物を疑う不透過像を認めた。下顎骨は右側第一小臼歯部の頬側皮質骨から臼後部舌側皮質骨に向かって斜走する骨折線を認めた。右側下顎側切歯、左側下顎第一大臼歯の歯冠破折と臨床診断し、静脈内鎮静下(ミダゾラム注)で気管支鏡を用い、気道異物を除去した。経過観察後、受傷3週間後に全身麻酔下に観血的整復固定術を行った。受傷9ヵ月経過したが開口時の左側偏位はあるが、開口量は58mmで日常生活に支障は認められない。

2003年1月〜2014年5月迄の11年4ヵ月に歯科・顎・口腔外科を受診した顎顔面骨折127例中、60歳以上の高齢者32例(25%)を対象に、60歳未満の症例と比較検討した。高齢者の性別は男性22例、女性10例、男女比2.2:1で男性が多く、年齢別では非高齢者群、高齢者群ともに男性が多いが高齢者群では他の年代よりも女性の割合が高かった。受傷原因は非高齢者群では交通外傷36例、転倒・転落36例、殴打11例、スポーツ9例、労働災害3例、高齢者群では転倒・転落23例、交通外傷7例、殴打2例と転落・転倒が大部分を占めた。骨折部位は非高齢者群では下顎骨骨折60例が最多で次いで上顎骨折11例、頬骨弓骨折10例の順で、複合骨折は14例であった。高齢者群は下顎骨骨折が17例と最多で、次いで上顎骨骨折5例、頬骨骨折5例で、複合骨折は5例を占めた。下顎骨の骨折線は非高齢者群では下顎前歯部26例、関節突起部22例、高齢者群では関節突起部9例、下顎前歯部7例であった。処置法は非高齢者群の非観血的処置39例、観血的処置28例、高齢者群では観血的処置13例、経過観察9例、非観血的処置28例、全身状態悪化で手術中止3例であった。受傷時刻は非高齢者群は夜間帯に受傷することが多かった。

上顎洞異物は外傷性と医原性に大別され、医原性異物においては歯科領域のものが多い。近年ではインプラントの普及に伴い、上顎洞へのインプラントの迷入報告例が増加している。われわれは、上顎洞内へ迷入したインプラントを内視鏡を併用して摘出した症例を報告する。患者は48歳女性で、近在歯科医院にてインプラント埋入術を施行した際にインプラント体およびカバースクリューが上顎洞に迷入し、摘出依頼にて当科紹介となった。パノラマX線写真にて右側上顎洞内の上方にインプラント体およびカバースクリューと思われるX線不透過像を認めた。CT写真においてインプラント体周囲の上顎洞粘膜は軽度肥厚しており、カバースクリューは篩骨洞への移行部付近に位置していた。全身麻酔下での処置を予定し術前検査を行ったところ、未治療の糖尿病が発覚したため、血糖管理を行ってから上顎洞異物摘出術を施行した。上顎洞粘膜は軽度肥厚し、異物の一部は篩骨洞への移行部付近に存在したが、内視鏡を用いることで視野を確保し犬歯窩から摘出した。上顎洞異物は、位置や大きさ、洞粘膜の状態、患者の全身状態等を正確に把握し、適切な治療法を選択することが重要であると考えられた。(著者抄録)

Tropomodulin1 (TMOD1), which regulates the length and depolymerization of actin filaments by binding to the pointed end of the actin filament, has been reported to be a powerful diagnostic marker for ALK-negative anaplastic large-cell lymphoma; however, little is known about the relevance of TMOD1 in the behavior of oral squamous cell carcinoma (OSCC). We evaluated TMOD1 expression in OSCC-derived cell lines and primary OSCC samples (n=200) using quantitative reverse transcriptase-polymerase chain reaction, immunoblotting and semi-quantitative immunohistochemistry. We also analyzed the clinical correlation between TMOD1 expression status and clinical parameters in patients with OSCC and performed a prospective study using 40 primary OSCC samples. TMOD1 expression was upregulated significantly (p&lt;0.05) in OSCC in vitro and in vivo compared with normal counterparts. TMOD1 expression also was correlated significantly (p=0.0199 and p=0.0064, respectively) with regional lymph node metastasis (RLNM) and 5-year survival rates. This prospective study also showed that high TMOD1 expression was seen in 12 (75%) of 16 cases in RLNM-positive patients and 9 (37.5%) of 24 cases in RLNM-negative patients. The current data provide the first evidence that TMOD1 expression is a critical biomarker for RLNM and prognosis of patients with OSCC.

Aryl hydrocarbon receptor nuclear translocator (ARNT) 2 is a transcriptional factor related to adaptive responses against cellular stress from a xenobiotic substance. Recent evidence indicates ARNT is involved in carcinogenesis and cancer progression; however, little is known about the relevance of ARNT2 in the behavior of oral squamous cell carcinoma (OSCC). In the current study, we evaluated the ARNT2 mRNA and protein expression levels in OSCC in vitro and in vivo and the clinical relationship between ARNT2 expression levels in primary OSCCs and their clinicopathologic status by quantitative reverse transcriptase-polymerase chain reaction, immunoblotting, and immunohistochemistry. Using ARNT2 overexpression models, we performed functional analyses to investigate the critical roles of ARNT2 in OSCC. ARNT2 mRNA and protein were down-regulated significantly (P < 0.05 for both comparisons) in nine OSCC-derived cells and primary OSCC (n=100 patients) compared with normal counterparts. In addition to the data from exogenous experiments that ARNT2-overexpressed cells showed decreased cellular proliferation, ARNT2-positive OSCC cases were correlated significantly (P < 0.05) with tumoral size. Since von Hippel-Lindau tumor suppressor, E3 ubiquitin protein ligase, a negative regulator of hypoxia-inducible factor (HIF1)-α, is a downstream molecule of ARNT2, we speculated that HIF1-α and its downstream molecules would have key functions in cellular growth. Consistent with our hypothesis, overexpressed ARNT2 cells showed down-regulation of HIF1-α, which causes hypofunctioning of glucose transporter 1, leading to decreased cellular growth. Our results proposed for the first time that the ARNT2 level is an indicator of cellular proliferation in OSCCs. Therefore, ARNT2 may be a potential therapeutic target against progression of OSCCs.

The endothelial-specific receptor, tyrosine kinase with immunoglobulin-like loops and epidermal growth factor homology domains-2 (Tie2) is a member of the tyrosine kinase family and is ubiquitous in normal tissues; however, little is known about the mechanisms and roles of Tie2 in oral squamous cell carcinomas (OSCCs). In the current study, we investigated the expression status of Tie2 in OSCCs by quantitative reverse transcriptase-polymerase chain reaction, immunoblotting, and immunohistochemistry and the functional mechanisms of Tie2 using its overexpressed OSCC (oeTie2) cells and Tie2 blocking by its antibody. We found that Tie2 expression was down-regulated significantly (p &lt; 0.05) in OSCCs compared with normal counterparts in vitro and in vivo. Interestingly, oeTie2 cells showed higher cellular adhesion (p &lt; 0.05) and lower cellular invasion (p &lt; 0.05) compared with control cells; whereas there was similar cellular proliferation in both transfectants. Furthermore, cellular adhesion was inhibited and invasion was activated by Tie2 function-blocking antibody (p &lt; 0.05), indicating that Tie2 directly regulates cellular adhesion and invasion. As expected, among the clinical variables analyzed, Tie2-positivity in patients with OSCC was correlated closely with negative lymph node metastasis. These results suggested for the first time that Tie2 plays an important role in tumor metastasis and may be a potential biomarker for OSCC metastasis.

63歳女性。近医の歯科にて左側頬粘膜部口内炎に対し歯科用炭酸ガスレーザーを照射後、左側頬部と顎下部に捻髪音を伴う腫脹が出現、腫脹は頸部まで拡大したため再度、近医の歯科を受診したが、皮下気腫を疑われ、著者らの施設へ紹介となった。受診時、口腔内所見では左側頬粘膜部にレーザー照射痕を認めたが、口腔清掃状態は良好でその他の異常は認められなかった。しかし、CT像では左側は上方が側頭隙、下方は鎖骨上、後方は後頭隙で、一方、右側は上方が乳様突起内側、下方は顎下部、後方は乳様突起まで及び、前方はオトガイ下隙にまで至る範囲で含気像と思われる所見が確認された。以後、感染予防および皮下気腫経過観察目的で入院となり、セフトリアキソンナトリウム(2g/日)の点滴投与を開始したところ、治療開始2日目から経時的に腫脹は軽減、治療開始14日目には消失した。更に治療開始21日目にはCT像にて皮下気腫の消失を確認し、その後は経過良好で終診とした。

Background Nucleolar and spindle-associated protein 1 (NUSAP1) is an important mitotic regulator. In addition to its crucial function in mitosis, NUSAP1 has recently received attention due to the interesting roles in carcinogenesis. The aim of this study was to reveal functional mechanisms of NUSAP1 in oral squamous cell carcinoma (OSCC). Methods mRNA and protein expression levels of NUSAP1 in 9 OSCC-derived cells were analyzed by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and immuno-blotting analyses. The correlation between the NUSAP1 expression profile and the clinicopathological factors was evaluated by immunohistochemistry (IHC) in clinical OSCC samples (n = 70). The NUSAP1 knockdown cells were established with short hairpin RNA (shRNA) in OSCC cells, and functional assays were performed using these cells. In addition to the evaluation of cellular proliferation and cell cycle, we also investigated the potential role of NUSAP1 in paclitaxel (PTX)-induced cellular responses. Results mRNA and protein expression of NUSAP1 were significantly up-regulated in OSCC-derived cells compared with human normal oral keratinocytes (P &lt; 0.05). IHC revealed that NUSAP-1 expression is closely associated with primary advanced T stage (P&lt;0.05). Suppression of NUSAP1 expression levels led to significant (P&lt;0.05) inhibition of cellular proliferation. Furthermore, apoptosis induced by PTX was enhanced in NUSAP1 knockdown OSCC cells. Conclusions NUSAP1 may be a crucial biomarker for OSCC. Moreover, down-regulated NUSAP1 expression suppresses tumor proliferation and also enhances anti-tumor effect of PTX by activating apoptotic pathways. Thus, the present study strongly suggests that regulating NUSAP1 expression should contribute to the therapy for OSCC.

口腔領域の悪性腫瘍に対しては手術療法が中心的な役割を果たしているが、種々の理由により手術以外の方法を選択する状況も見受けられる。本研究では、手術以外の治療方法により加療した口腔領域悪性腫瘍について臨床的な検討を行った。2000年1月から2012年12月までの12年間に当科を受診した口腔扁平上皮癌症例は445例であり、そのうち非手術症例を選択した患者は69例(15.5%)であった。手術以外の治療法を選択した理由としては、高度進展、患者意思、高齢、全身状態不良、家族選択であった。治療法の内訳は選択的動注化学療法併用放射線療法34例(49.3%)、化学放射線療法15例(21.7%)、放射線療法12例(17.4%)、化学療法6例(8.6%)、終末期医療2例(3.0%)であった。手術症例との比較では、5年累積生存率は、非手術症例が28.3%、手術症例が85.9%、非手術症例の生存期間中央値は25.8ヵ月であった。放射線療法の新たな選択肢としてサイバーナイフによる放射線治療も行われるようになっており、非手術症例でサイバーナイフを施行した成績は、完全奏効率(CR率)は16.6%、奏効率(CR+PR率)50.0%、疾患制御率(CR+PR+SD率)100%で局所制御に有用であることが示唆された。今回の調査では、治療方法としては全身状態が許されれば可能な限り手術療法を行った方がよいが、手術以外の治療としては、選択的動注化学療法併用放射線療法は有効な方法であることが示され、サイバーナイフによる局所制御も有望な治療方法であることが示唆された。(著者抄録)

抗癌剤や放射線耐性に関しては、感受性細胞と耐性細胞の組み合わせを用いたマイクロアレイ解析により、耐性細胞において高発現した遺伝子群を耐性候補遺伝子とした。これらの候補遺伝子に関し、遺伝子学的機能解析と臨床サンプルを用いた免疫染色法による機能解析を行ない、癌治療における標的候補遺伝子を絞り込んだ。また、転移抑制に関しては、2次元タンパク泳動-質量解析法で、癌特異的に発現低下したタンパクを検出・同定した。このタンパクの発現状態と頸部リンパ節転移の有無に関する臨床症例による比較検討と、発現ベクター導入による遺伝子強制発現実験、in vivo転移実験により転移制御因子であることを明らかにし、癌治療における標的候補遺伝子を同定した。ドラッグ・リポジショニング手法を用いて、現在他の疾患の治療薬として使用されている医薬品、あるいは過去に医薬品開発過程で放棄された化合物のリストから、これらの標的候補遺伝子に対する発現制御薬を検索し、in vitro、in vivoによる機能解析により、薬剤の作用を確認した。これらの薬剤は、臨床の場で実際に使用可能な治療薬候補として期待できるものと考えられる。(著者抄録)

Circulating tumor cells (CTCs) and/or their relating molecules are promising determinants during the course of cancer treatment, especially for post-therapeutic monitoring. We recently reported the clinical relevance of detecting circulating tumor-associated mutant mitochondrial DNAs (mut-mtDNAs) at three different regions including the displacement loop, 12S-rRNA and 16S-rRNA in oral squamous cell carcinomas (OSCCs). In the present study, to further investigate if the other mut-mtDNAs have novel efficiency for detecting potential tumoral micrometastasis, mut-mtDNAs on the ND2 and ND3 regions of the genome in 240 clinical samples from patients with OSCC were assessed in vitro and in vivo by quantitative real-time PCR combined with high-resolution melting curve analysis. Furthermore, the clinical relevance was evaluated by the area under the receiver operating characteristic curve (AUC) analysis. Three discrete sequence variations were identified in OSCC derived cell lines at the regions of ND2 (T:A to C:G at position 5108) and ND3 (A:T to G:C at position 10397 and C:G to T:A at position 10400), whereas no mutation was observed in normal control human normal oral keratinocytes. In OSCC patients examined, the presence of mut-mtDNAs in serum during the postoperative period accurately predicted poor prognoses (ND2 AUC, 0.761; ND3 AUC, 0.704). The data presented here provide a novel approach for detecting the circulating mut-mtDNAs that are promising molecular markers for evaluating tumoral micrometastasis in OSCCs.

Background Semaphorins (SEMAs) consist of a large family of secreted and membrane-anchored proteins that are important in neuronal pathfinding and axon guidance in selected areas of the developing nervous system. Of them, SEMA7A has been reported to have a chemotactic activity in neurogenesis and to be an immunomodulator; however, little is known about the relevance of SEMA7A in the behaviors of oral squamous cell carcinoma (OSCC). Methods We evaluated SEMA7A expression in OSCC-derived cell lines and primary OSCC samples using quantitative reverse transcriptase-polymerase chain reaction, immunoblotting, and semiquantitative immunohistochemistry (sq-IHC). In addition, SEMA7A knockdown cells (5hSEMA7A cells) were used for functional experiments, including cellular proliferation, invasiveness, and migration assays. We also analyzed the clinical correlation between SEMA7A status and clinical behaviors in patients with OSCC. Results SEMA7A mRNA and protein were up-regulated significantly (P&lt;0.05) in OSCC-derived cell lines compared with human normal oral keratinocytes. The shSEMA7A cells showed decreased cellular growth by cell-cycle arrest at the G1 phase, resulting from up-regulation of cyclin-dependent kinase inhibitors (p21c1P1 and p27KiP1) and down-regulation of cyclins (cyclin Dl, cyclin E) and cyclin-dependent kinases (CDK2, CDK4, and CDK6); and decreased invasiveness and migration activities by reduced secretion of matrix metalloproteases (MMPs) (MMP-2, proMMP-2, pro-MMP-9), and expression of membrane type 1MMP (MT1-MMP). We also found inactivation of the extracellular regulated kinase 1/2 and AKT pathways, an upstream molecule of cell-cycle arrest at the G1 phase, and reduced secretion of MMPs in shSEMA7A cells. sq-IHC showed that SEMA7A expression in the primary OSCCs was significantly (P = 0.001) greater than that in normal counterparts and was correlated with primary tumoral size (P = 0.0254) and regional lymph node metastasis (P = 0.0002). Conclusion Our data provide evidence for an essential role of SEMA7A in tumoral growth and metastasis in OSCC and indicated that SEMA7A may play a potential diagnostic/therapeutic target for use in patients with OSCC.

Kinesin family member 14 (KIF14), a microtubule-based motor protein, plays an important role in chromosomal segregation, congression, and alignment. Considerable evidence indicates that KIF14 is involved in cytokinesis, although little is known about its role in oral squamous cell carcinomas (OSCCs). In the current study, we functionally and clinically investigated KIF14 expression in patients with OSCC. Quantitative reverse transcriptase-polymerase chain reaction and immunoblotting analyses were used to assess the KIF14 regulatory mechanism in OSCC. Immunohistochemistry (IHC) was performed to analyze the correlation between KIF14 expression and clinical behavior in 104 patients with OSCC. A KIF14 knockdown model of OSCC cells (shKIF14 cells) was used for functional experiments. KIF14 expression was up-regulated significantly (. P&lt 0.05) in OSCCs compared with normal counterparts in vitro and in vivo. In addition, shKIF14 cells inhibited cellular proliferation compared with control cells by cell-cycle arrest at the G2/M phase through up-regulation of G2 arrest-related proteins (p-Cdc2 and cyclin B1). As expected, IHC data from primary OSCCs showed that KIF14-positive patients exhibited significantly (. P&lt 0.05) more larger tumors compared with KIF14-negative patients. The current results suggest for the first time that KIF14 is an indicator of tumoral size in OSCCs and that KIF14 might be a potential therapeutic target for development of new treatments for OSCCs.

Persephin (PSPN) is a neurotrophic factor of the glial cell line-derived neurotrophic factor (GDNF) family that promotes survival of multiple populations of neurons. Little is known about the relevance of PSPN in human malignancy including oral squamous cell carcinoma (OSCC). This study was undertaken to evaluate PSPN mRNA and protein expression by analyzing cellular proliferation and the cell cycle in PSPN knockdown cells in vitro. PSPN mRNA and protein were significantly (P&lt;0.05) up-regulated in OSCC-derived cells compared with human normal oral keratinocytes (n=7). Cellular proliferation decreased significantly (P&lt;0.05) in PSPN knockdown cells with reduced receptor tyrosine kinase (RTK) signaling, and cell-cycle arrest at the G1 phase resulted from up-regulation of the cyclin-dependent kinase inhibitors (p21(Cip1), p27(Kip1), p15(INK4B), and p16(INK4A)). Furthermore, the PSPN protein expression in 101 primary OSCCs was significantly (P&lt;0.05) higher than in normal counterparts. Among the clinical variables analyzed, overexpression of PSPN also was related closely (P&lt;0.05) to tumoral size. Our results suggested that PSPN is a possible key regulator of OSCC progression via PSPN-RET-mitogen-activated protein kinase activation and that PSPN overexpression may have diagnostic potential for OSCC. (c) 2013 Wiley Periodicals, Inc.