CRNDE promotes cell tongue squamous cell carcinoma cell growth and invasion through suppressing miR-384.

Tongue squamous cell carcinoma (TSCC) is the most common type of oral cancer and is an aggressive head and neck malignancy. Increasing studies have demonstrated that long noncoding RNAs (lncRNAs) play important roles in diverse biological cell processes, such as cell development, fate decisions, cell differentiation, cell migration, and invasion. In our study, we showed that long noncoding RNA colorectal neoplasia differentially expressed (CRNDE) expression was upregulated in TSCC cell lines and tissues. Overexpression of CRNDE increased the TSCC cell proliferation, cell cycle, and cell invasion. Moreover, ectopic expression of CRNDE inhibited the miR-384 expression in the SCC1 cell and increased the Kirsten Ras (KRAS), cell division cycle 42, and insulin receptor substrate 1 expression, which were the direct target genes of miR-384. We demonstrated that the miR-384 expression was downregulated in the TSCC samples compared with the paired adjacent nontumor samples. The expression of CRNDE was negatively correlated with the expression of miR-384 in the TSCC samples. Overexpression of miR-384 suppressed TSCC cell proliferation, cell cycle, and invasion. Furthermore, we demonstrated that CRNDE promoted TSCC cell proliferation and invasion through inhibiting miR-384 expression. These results suggested that CRNDE acts as an oncogene in the development of TSCC, which partially occurs through inhibiting miR-384 expression.

The Neuropilins and Their Ligands in Hematogenous Metastasis of Salivary Adenoid Cystic Carcinoma-An Immunohistochemical Study.

PURPOSE: We investigated the expression of neuropilin-1 (NRP1), neuropilin-2 (NRP2), vascular endothelial growth factor-A (VEGF-A), semaphorin-3A (Sema-3A), and semaphorin-3F (Sema-3F) in normal salivary gland (NSG) tissue, nonmetastatic salivary adenoid cystic carcinoma (SACC), and metastatic SACC to better understand their role in intratumoral angiogenesis and hematogenous metastasis of SACC. PATIENTS AND METHODS: The study included 60 SACC patients, equally divided between nonmetastatic SACC and metastatic SACC. We used 30 NSG samples as the control. The expression of cytokines was studied by immunohistochemistry and compared using the integrated optical density. The relationship between NRP1, NRP2, VEGF-A, and Sema-3A expression and microvessel density (MVD) was analyzed in the 3 groups. RESULTS: In metastatic SACC, the expression levels of NRP1 and VEGF-A were significantly greater than those in nonmetastatic SACC and NSG. The expression of Sema-3A and Sema-3F was significantly lower in metastatic SACC than that in nonmetastatic SACC and NSG (P < .0001). No significant differences were found in NRP2 expression among the 3 groups (P = .43). The MVD of metastatic SACC was significantly greater than that of nonmetastatic SACC and NSG (P < .0001). However, the lymphatic vessel density of the 3 groups was not significantly different statistically. The relationship between MVD and NRP1 or VEGF-A showed a significant positive correlation (P < .0001, for both). However, a significant negative correlation was found between the MVD and Sema-3A or Sema-3F expression (P < .0001, for both). CONCLUSIONS: Hematogenous metastasis of SACC is correlated with high expression of NRP1 and VEGF-A and low expression of Sema-3A and Sema-3F. The increased numbers of microvessels induced by VEGF-A signaling, combined with NRP1, could be one of the key reasons leading to the enhanced hematogenous metastasis in SACC.

Sympathetic innervation contributes to perineural invasion of salivary adenoid cystic carcinoma via the ß2-adrenergic receptor.

PURPOSE: Perineural invasion (PNI) is reported to correlate with local recurrence and poor prognosis of salivary adenoid cystic carcinoma (SACC). However, the pathogenesis of PNI remains unclear. The aims of this study were to investigate the correlation between sympathetic innervation and SACC PNI and to elucidate how the sympathetic neurotransmitter norepinephrine (NE) regulates the PNI process. MATERIALS AND METHODS: Sympathetic innervation and ß2-adrenergic receptor (ß2-AR) expression in SACC tissues were evaluated by immunohistochemistry. The NE concentrations in SACC tissues and dorsal root ganglia (DRG) coculture models were measured by ELISA. ß2-AR expression in SACC cells was detected by performing quantitative real-time polymerase chain reaction (qRT-PCR) and immunofluorescence assay. SACC cells were treated with NE, the nonselective α-AR blocker phentolamine, the ß2-AR antagonist ICI118,551, or were transfected with ß2-AR small interfering RNA (siRNA). Proliferation was evaluated in methyl thiazolyl tetrazolium assay, and migration was evaluated in Transwell assay and wound-healing assay. PNI was tested through both Transwell assay and a DRG coculture model. The expressions of epithelial-mesenchymal transition (EMT) markers and matrix metalloproteinases (MMPs) were measured by performing qRT-PCR and Western blot assay. RESULTS: Sympathetic innervation and ß2-AR were highly distributed in SACC tissues and correlated positively with PNI (P=0.035 and P=0.003, respectively). The sympathetic neurotransmitter NE was overexpressed in SACC tissues and DRG coculture models. Exogenously added NE promoted proliferation, migration, and PNI of SACC cells via ß2-AR activation. NE/ß2-AR signaling may promote proliferation, migration, and PNI by inducing EMT and upregulating MMPs. However, ß2-AR inhibition with either an antagonist or siRNA abrogated NE-induced PNI. CONCLUSION: Collectively, our findings reveal the supportive role of sympathetic innervation in the pathogenesis of SACC PNI and suggest ß2-AR as a potential therapeutic target for treating PNI in SACC.

Nomograms to estimate long-term overall survival and tongue cancer-specific survival of patients with tongue squamous cell carcinoma.

The aim of this study was to construct nomograms to predict long-term overall survival (OS) and tongue cancer-specific survival (TCSS) of tongue squamous cell carcinoma (TSCC) patients based on clinical and tumor characteristics. Clinical, tumor, and treatment characteristics of 12,674 patients diagnosed with TSCC between 2004 and 2013 were collected from the Surveillance, Epidemiology, and End Results database. These patients were then divided into surgery and nonsurgery cohorts, and nomograms were developed for each of these groups. The step-down method and cumulative incidence function were used for model selection to determine the significant prognostic factors associated with OS and TCSS. These prognostic variables were incorporated into nomograms. An external cohort was used to validate the surgery nomograms. Seven variables were used to create the surgery nomograms for OS and TCSS, which had c-indexes of 0.709 and 0.728, respectively; for the external validation cohort, the c-indexes were 0.691 and 0.711, respectively. Nine variables were used to create the nonsurgery nomograms for OS and TCSS, which had c-indexes of 0.750 and 0.754, respectively. The calibration curves of the 5- and 8-year surgery and nonsurgery nomograms showed excellent agreement between the probabilities and observed values. By incorporating clinicopathological and host characteristics in patients, we are the first to establish nomograms that accurately predict prognosis for individual patients with TSCC. These nomograms ought to provide more personalized and reliable prognostic information, and improve clinical decision-making for TSCC patients.

Overexpression of KAI1/CD82 suppresses in vitro cell growth, migration, invasion and xenograft growth in oral cancer.

KAI1/CD82 is a metastatic suppressor gene in human prostate cancer and several other types of cancer in humans. The present study aimed to examine the role of the overexpression of KAI1 in the progression of oral cancer. Human KAI1/CD82 cDNA was transfected into OSCC­15 and 293T cell lines, and its effects on OSCC­15 cell proliferation, invasion and apoptosis were assessed by performing a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, Matrigel invasion and Annexin V­FITC staining, respectively. In addition, a xenograft model was used to assess the effect of KAI1/CD82 on the in vivo growth of tumors. The overexpression of KAI1/CD82 inhibited the proliferation and invasion of OSCC-15 cells. It also enhanced the apoptotic rate of the OSCC­15 cells. Furthermore, the overexpression of KAI1/CD82 inhibited tumor growth in the xenograft model. The results demonstrated that the overexpression of KAI1/CD82 significantly inhibited the proliferation and invasion of human oral cancer cells, and inhibited tumor growth in the xenograft model. Therefore, KAI1/CD82 may be considered as a potential therapeutic target in oral cancer.

Nomograms for predicting long-term overall survival and cancer-specific survival in patients with major salivary gland cancer: a population-based study.

In this study, we aimed to develop and validate nomograms for predicting long-term overall survival (OS) and cancer-specific survival (CSS) in major salivary gland cancer (MSGC) patients. These nomograms were developed using a retrospective cohort (N=4218) from the Surveillance, Epidemiology, and End Results (SEER) database, and externally validated using an independent data cohort (N=244). We used univariate, and multivariate analyses, and cumulative incidence function to select the independent prognostic factors of OS and CSS. Index of concordance (c-index) and calibration plots were used to estimate the nomograms' predictive accuracy. The median follow-up period was 34 months (1-119 months). Of 4218 MSGC patients, 1320 (31.3%) died by the end of the follow-up; of these 1320 patients, 883 (20.9%) died of MSGC. The OS nomogram, which had a c-index of 0.817, was based on nine variables: age, sex, tumor site, tumor grade, surgery performed, radiation therapy and TNM classifications. The CSS nomogram, which had a c-index of 0.829, was based on the same nine variables plus race. External validation c-indexes were 0.829 and 0.807 for OS and CSS, respectively. Based on SEER database, we have developed nomograms predicting five- and eight-years OS and CSS for MSGC patients with perfect accuracy. These nomograms will help clinicians customize treatment and monitoring strategies in MSGC patients.

The role of perineural invasion on head and neck adenoid cystic carcinoma prognosis: a systematic review and meta-analysis.

OBJECTIVE: We performed a systematic review to assess the prognostic value of perineural invasion (PNI) for patients with head and neck adenoid cystic carcinoma. STUDY DESIGN: A literature search of MEDLINE and EMBASE was used to identify relevant literature up to December 2015. The primary outcomes of interest were overall survival, disease-free survival, and locoregional control. Study information and hazard ratios (HRs) were extracted, and HRs were pooled using the Mantel-Haenszel fixed-effects model and the DerSimonian and Laird random-effects model according to heterogeneity. RESULTS: Twenty-two studies and 1332 patients were included in this study. The PNI ratio was 43.2%. PNI was at increasing risk for overall survival (HR = 2.98; 95% confidence interval [CI] 2.00-4.46), disease-free survival (HR = 1.88; 95% CI, 1.42-2.49), and locoregional control (HR = 2.15; 95% CI, 1.48-3.13) with statistical significance. CONCLUSIONS: PNI is an independent factor for poor prognosis in patients with head and neck adenoid cystic carcinoma. Moreover, PNI poses a significantly higher threat to male patients and younger patients.

Nomograms predicting long-term overall survival and cancer-specific survival in head and neck squamous cell carcinoma patients.

This study aimed to develop nomograms to predict long-term overall survival and cancer-specific survival in patients with head and neck squamous cell carcinoma (HNSCC). We conducted prognostic analyses and developed nomograms predicting survival outcome using HNSCC patient data collected from the Surveillance, Epidemiology and End Results (SEER) program of the National Cancer Institute. An external dataset of 219 patients was used to validate the nomograms. Of 36,179 HNSCC patients, 9,627 (26.6%) died from HNSCC and 4,229 (11.7%) died from other causes. Median follow-up was 28 months (1-107 months). Nomograms predicting overall survival (OS) and cancer-specific survival (CSS) were developed according to 10 clinicopathologic factors (age, race, sex, tumor site, tumor grade, surgery, radiotherapy and TNM stage), with concordance indexes (C-indexes) of 0.719 and 0.741, respectively. External validation C-indexes were 0.709 and 0.706 for OS and CSS, respectively. Our results suggest that we successfully developed nomograms predicting five- and eight-year HNSCC patient OS and CSS with high accuracy. These nomograms could help clinicians tailor surgical, adjuvant therapeutic and follow-up strategies to more effectively treat HNSCC patients.

p12CDK2-AP1 interacts with CD82 to regulate the proliferation and survival of human oral squamous cell carcinoma cells.

p12 cyclin-dependent kinase 2 (CDK2)-associating protein 1 (p12CDK2-AP1) has been demonstrated to negatively regulate the activity of CDK2. However, the underlying molecular mechanism remains largely unknown. We aimed to determine the potential binding proteins of p12CDK2-AP1 and to elucidate the role of p12CDK2-AP1 in the regulation of the proliferation, invasion, apoptosis, and in vivo growth of human oral squamous cell carcinoma cells. The protein-protein interaction was predicted using computational decision templates. The predicted p12CDK2­AP1 interacting proteins were overexpressed in human oral squamous cell carcinoma OSCC-15 cells, and the protein binding was examined using co-precipitation (Co-IP). Cell proliferation and invasion were determined via MTT assay and Transwell system, respectively. Cell apoptosis was evaluated using Annexin V-FITC/PI double staining followed by flow cytometric analysis. The in vivo growth of OSCC-15 cells was examined in nude mouse tumor xenografts. We found that overexpression of either p12CDK2-AP1 or CD82 significantly suppressed the proliferation and invasion but promoted the apoptosis of OSCC-15 cells (P<0.05). Importantly, combined overexpression of p12CDK2-AP1 and CD82 showed synergistic antitumor activity compared with the overexpression of a single protein alone (P<0.05). Additionally, the simultaneous overexpression of p12CDK2-AP1 and CD82 significantly suppressed the in vivo tumor growth of OSCC-15 cells in nude mice compared with the negative control (P<0.05). Our findings indicate that p12CDK2-AP1 interacts with CD82 to play a functional role in suppressing the in vitro and in vivo growth of OSCC-15 cells.

[Clinical research for the treatment of temporomandibular joint injury based on three-dimensional digital technology].

OBJECTIVE: To investigate the accurate and individual treatment of temporomandibular joint(TMJ) injury based on 3D digital technology. METHODS: Maxillo-mandibular model was made using rapid prototyping technology based on the pre-operation 3D CT results. According to the 3D digital measurement results, TMJ concepts were ordered and the prosthesis was used to simulate the replacement surgery on the model. Then the joint replacement surgery was performed afterward. RESULTS: (1)After total replacement of TMJ, no pain happened and mouth open was not limited. Three months later, the joint position was normal and stable. The month open width was 4 cm. (2)After condyle replacement, primary healing was achieved with complete survival of bone graft. No edema was seen. Symmetric facial appearance was satisfactory. CONCLUSIONS: Bilateral individual prosthesis for total TMJ or condyle replacement is an ideal method for TMJ injury.

Interaction between p12CDK2AP1 and a novel unnamed protein product inhibits cell proliferation by regulating the cell cycle.

Human p12CDK2AP1 protein is encoded by the cyclin­dependent kinase 2­associated protein 1 (CDK2AP1) gene. This protein suppresses cell growth, differentiation and angiogenesis in numerous types of carcinoma by interacting with certain cell cycle proteins, including CDK2 and DNA polymerase α/primase. p12CDK2AP1 exerts its functions predominantly through protein­protein interactions. Therefore, the identification of other p12CDK2AP1­interacting proteins may clarify its role in cell cycle regulation and carcinogenesis. The aim of this study was to identify additional p12CDK2AP1­interacting proteins. A novel unnamed protein product (UPP, BC006130) was identified through using a yeast two­hybrid system. The interaction of p12CDK2AP1 with the UPP was further verified by glutathione S-transferase pull­down and co­immunoprecipitation experiments in vitro. The qPCR results following overexpression and siRNA assays demonstrated that the expression levels of the UPP were mediated by the CDK2AP1 gene. Furthermore, overexpression of the UPP gene was shown to shorten the length of the G2/M phase of the cell cycle in normal and tumor cell lines in a flow cytometry assay. The results of human tumor xenografts experiments in Balb/c nude mice indicated that stable transfection with the UPP gene was able to inhibit tumor cell proliferation in vivo. Overall, this study identified and characterized a novel interactive protein of p12CDK2AP1, which may inhibit cell proliferation by mediating the cell cycle. It expands the understanding of the mechanisms of p12CDK2AP1 and its potential as a cancer therapeutic target.

Silencing P12CDK²AP¹ with a lentivirus promotes HaCaT cell proliferation.

The tumor suppressor P12CDK2AP1 negatively regulates cyclin-dependent kinase 2 (CDK2) activities and suppresses DNA replication. Notably, P12CDK2AP1 is known to be downregulated in head and neck squamous cell carcinomas (HNSCCs). Silencing of specific gene expression by small interfering RNAs (siRNAs) or short hairpin RNAs (shRNAs) using expression vectors and retroviruses has become a powerful tool for the genetic analysis of mammalian cells. In the present study, we utilized lentivirus­mediated shRNA for functional gene knockdown in normal human skin keratinocytes (HaCaT) cells in order to assess the potential role of P12CDK2AP1 in HNSCCs. Lentivirus­mediated RNA interference (RNAi) effectively reduced endogenous P12CDK2AP1 expression in HaCaT cells and significantly promoted HaCaT cell proliferation in vitro. Lentiviral vectors have the ability to infect dividing and non-dividing cells as well as to achieve long­term multilineage gene expression. Thus, additional studies are needed to investigate the use of such vectors as a therapeutic tool for the delivery of siRNAs.