Poly Adenylate Binding Protein Interacting protein 1 (PAIP1) plays a critical role in translation initiation and is associated with the several cancer types. However, its function and clinical significance have not yet been described in oral squamous cell carcinoma (OSCC) and its associated features like lymph node metastasis (LNM). Here, we used the data available from Gene Expression Omnibus (GEO), The Cancer Genome Atlas (TCGA), and Clinical Proteomic Tumor Analysis Consortium (CPTAC) to analyze PAIP1 expression in oral cancer. The publicly available data suggests that PAIP1 mRNA and protein levels were increased in OSCC. The high PAIP1 expression was more evident in samples with advanced stage, LNM, and worse pattern of invasion. Moreover, the in vitro experiments revealed that PAIP1 knockdown attenuated colony forming, the aggressiveness of OSCC cell lines, decreasing MMP9 activity and SRC phosphorylation. Importantly, we found a correlation between PAIP1 and pSRC through the analysis of the IHC scores and CPTAC data in patient samples. Our findings suggest that PAIP1 could be an independent prognostic factor in OSCC with LNM and a suitable therapeutic target to improve OSCC patient outcomes.
Carcinoma, Squamous Cell/genetics* ; Head and Neck Neoplasms ; Humans ; Lymphatic Metastasis ; Mouth Neoplasms/pathology* ; Peptide Initiation Factors/metabolism* ; Prognosis ; Proteomics ; RNA-Binding Proteins/metabolism* ; Squamous Cell Carcinoma of Head and Neck

OBJECTIVE: To explore the association between hypoxia-inducible factor 1α (HIF-1α) expression and lymph node metastasis in oral squamous cell carcinoma (OSCC). METHODS: Tumor specimens from 125 patients with histologically-proven, surgically-treated OSCC were examined by immunohistochemical staining for expression of HIF-1α. The patients were divided into two groups by the expression of HIF-1α, high expression of HIF -1α group (H-group) and low expression of HIF-1α group (L-group). The main assessment parameters were lymph node metastasis rate and disease-specific survival (DSS). The lymph node metastasis rate and clinicopathologic features were compared using Mann-Whitney test. The Kaplan-Meier curve was generated for each group and compared using the log-rank test. Cox proportional hazard models were utilized for multivariate analyses of HIF-1α expression and other baseline factors with DSS. All calculations and analyses were performed using the SPSS 17.0 software package. RESULTS: The protein expression levels of HIF-1α were up-regulated in OSCC and two patients were unable to evaluate. There were 48 patients in L-group and 75 patients in H-group. Lymph node metastasis rate was 37.5% (18/48) for L-group and 58.7% (44/75) for H-group (P=0.027). Expression of HIF-1α was significantly correlated with lymph node metastasis. The patients of L-group had a significantly better DSS than the patients of H-group (70.8% vs. 46.7%, P=0.005), while the patients of L-group had a significantly better disease-free survival (DFS) than the patients of H-group (60.4% vs. 36.0%, P=0.009) by Kaplan-Meier method. A multivariate survival analysis also showed that HIF-1α expression (HR=2.164, 95%CI: 1.150-4.074, P=0.017) and T-stage (HR=1.387, 95%CI: 1.066-1.804, P=0.015) both were the independent factors associated with prognosis. CONCLUSION HIF-1α expression is significantly correlated with lymph node metastasis in OSCC. HIF-1α expression is an independent predictive factor for prognosis of OSCC patients, and may serve as a potential biomarker for molecular diagnosis and targeted therapy in future.
Biomarkers/metabolism* ; Carcinoma, Squamous Cell/pathology* ; Humans ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Immunohistochemistry ; Lymph Nodes ; Lymphatic Metastasis/genetics* ; Mouth Neoplasms/pathology* ; Prognosis

OBJECTIVETo investigate the expression of long non-coding RNA(lncRNA) colon cancer associated transcript 2(CCAT2) and its association with clinicopathologic features in oral squamous cell carcinoma(OSCC).

METHODSThe expression of lncRNA was detected with microarray assay in three samples of OSCC tumor and matched adjacent tissues. The profiles of lncRNAs in OSCC tissues were identified. The CCAT2 expression was evaluated by real-time quantitative PCR(RT-qPCR) in 86 OSCC tumor samples and matched adjacent tissues. The relationship between the expression of CCAT2 and its clinicopathologic features of OSCC was analyzed. Tumor cell proliferation was assessed following siRNA knockdown of CCAT2 by using the CCK-8 kits.

RESULTSA total of 1 685 lncRNA expressed in OSCC tumor samples and matched adjacent tissues were identified using microarray assay(P<0.05). RT-qPCR showed that the expression of CCAT2 was significantly higher in OSCC than that in adjacent tissues(P< 0.01). High CCAT2 expression was associated with cell differentiation and pathological stage of OSCC. CCAT2 expression in low-differentiated OSCC was significantly higher than that in high-differentiated cancer (P=0.015). In addition, CCAT2 level in stage Ⅲ/Ⅳ OSCC was significantly higher than that in stage Ⅰ/Ⅱ cancer (P=0.022). Furthermore, inhibition of CCAT2 expression suppressed the proliferation of human tongue carcinoma Tca8113 cells.

CONCLUSIONSAbnormal expression of lncRNA may be involved in the development of OSCC. Up-regulation of CCAT2 expression in tumor tissue might act as an oncogene and promote the development of OSCC.

Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Proliferation ; Gene Expression Profiling ; Humans ; Mouth ; metabolism ; Mouth Neoplasms ; genetics ; pathology ; RNA, Long Noncoding ; analysis ; metabolism ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Up-Regulation

This study is to investigate the antitumor activity of CIP-36 on multidrug resistant human oral squamous carcinoma cell line (KBV200 cells) in vitro and the possible anticancer mechanisms. MTT assay, Hoechst fluorescein stain, RT-PCR and immunohistochemistry were carried out on KBV200 and KB cells. The growth of many tumor cells was obviously inhibited by CIP-36, especially the multidrug resistant cells KBV200. Obvious apoptosis could be observed in the Hoechst 33342 staining experiments. The results of RT-PCR showed that the levels of p53, p21, caspase-3 and bax mRNA increased, and meanwhile the expression of mdr-1 and bcl-2 mRNA decreased in a dose-dependent manner. The data were significantly different from that of vehicle. The expression of P-gp significantly decreased with the increasing dosage of CIP-36 examined by immunohistochemistry. It can be concluded that CIP-36 could change resistance-related genes and proteins to overcome multidrug resistance in the KBV200 cell line.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Caspase 3 ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; KB Cells ; Mouth Neoplasms ; metabolism ; pathology ; Podophyllotoxin ; administration & dosage ; analogs & derivatives ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins p21(ras) ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo investigate the expression of galectin-1 in oral squamous cell carcinoma(OSCC) and its clinical significance.

METHODSDetection of the mRNA and protein expression of galectin-1 in the in vitro cellular carcinogenesis model of OSCC, OSCC cell lines and tissue specimens from 30 primary OSCC patients were performed using real-time polymerase chain reaction (PCR), Western blotting and immunohistochemistry, respectively.

RESULTSThe value of galectin-1 mRNA and protein level in human immortalized oral epithelia cell (HIOEC) cell was 0.071 ± 0.023, 0.118 ± 0.046, Compared with the HIOEC, galectin-1 mRNA level and protein expression were increased significantly in all the cell lines (0.141 ± 0.049, 0.504 ± 0.33) (P < 0.01). The levels of mRNA and protein expression of galectin-1 were significantly higher in the cancerous tissue (0.059 ± 0.034, 1.5 ± 0.68) than in the normal adjacent tissues (0.029 ± 0.012, 0.4 ± 0.56) (P < 0.01).

CONCLUSIONSThe expression of galectin-1 gene up-regulated in carcinogenesis process of OSCC significantly may be related to the tumorigenesis and development of OSCC, which illustrates its potential clinical application as tumor marker for early diagnosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Transformed ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Epithelial Cells ; cytology ; metabolism ; Female ; Galectin 1 ; genetics ; metabolism ; Humans ; Male ; Middle Aged ; Mouth Mucosa ; metabolism ; pathology ; Mouth Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Up-Regulation

PURPOSE: Surgical therapy is the primary treatment for oral cancer, but it can cause facial distortion. Therefore, if anticancer drugs are effective against oral cancer, they may be used preferentially. However, oral squamous carcinoma cells (OSCCs) are resistant to these drugs, so finding a way to enhance the sensitivity of these cells to anticancer drugs is important. The bacterial protein azurin is known to selectively enter cancer cells and induce apoptosis. In this study, we show the anticancer effect of azurin in OSCC. MATERIALS AND METHODS: OSCC cell line (YD-9) was subjected to azurin treatment. Cell viability, morphology and protein expression levels were monitored after treatment of azurin. Cells were also subjected to combination treatment of azurin with either 5-fluorouracil or etopside. RESULTS: Azurin-treated cells showed decreased cell viability accompanied by apoptotic phenotypes including morphological change, DNA breakage, and increases in p53 and cyclin B1 protein levels. Combination treatment of azurin with other anti-tumor agents caused an increase in sensitivity to anticancer drugs in azurin-treated YD-9 cells. CONCLUSION: Azurin has a strong synergistic anticancer effect on oral cancer cells when it is used along with anticancer drugs.
Antineoplastic Agents/*administration & dosage ; Apoptosis/drug effects ; Azurin/*administration & dosage/genetics ; Carcinoma, Squamous Cell/*drug therapy/metabolism/pathology ; Cell Line, Tumor ; Cyclin B1/metabolism ; Drug Synergism ; Etoposide/administration & dosage ; Fluorouracil/administration & dosage ; Humans ; Mouth Neoplasms/*drug therapy/metabolism/pathology ; Tumor Suppressor Protein p53/metabolism

OBJECTIVETo study the mechanism underlying the effect of combined use of cyclonpamine and hydroxycamptothecin in inducing the apoptosis of human oral squamous cell carcinoma cell line (OSCC) HSQ-89.

METHODSCCK8 assay was used to investigate the inhibitory effect of cyclopamine on HSQ-89 cells. Flow cytometry (FCM) was employed to examine the cell apoptosis following combined treatment with cyclonpamine and hydroxycamptothecin. Reverse transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expressions of Bcl-2, Bcl-xl, and Bid in HSQ-89 cells after the treatments.

RESULTSCombined treatment with cyclonpamine and hydroxycamptothecin significantly inhibited the cell proliferation compared with hydroxycamptothecin treatment alone, also resulting in a significantly higher apoptosis rate of the cells (P<0.05). The mRNA level of Bcl-2 was significantly decreased after the treatments, especially after the combined treatment. Cyclopamine produced no significant effect on the mRNA levels of Bcl-xl and Bid in the cells.

CONCLUSIONThe combined use of cyclopamine and hydroxycamptothecin significantly down-regulates the expression on of bcl-2 to induce the apoptosis of human OSCC cell line HSQ-89.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Carcinoma, Squamous Cell ; pathology ; Cell Line, Tumor ; Drug Synergism ; Humans ; Mouth Neoplasms ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Veratrum Alkaloids ; pharmacology

OBJECTIVETo study the expression of livin at the invasive tumor front of oral squamous cell carcinoma.

METHODSForty-eight samples of oral squamous cell carcinoma were graded according to invasive front gading (IFG). The expression of livin was evaluated at the ITF and other parts of the same tumor using immunohistochemistry.

RESULTSSignificant difference in the pathological grades was found between the ITF and the other parts of oral squamous cell carcinoma (P<0.01). The expression of livin at the ITF was significantly stronger than that in the other regions (P<0.01). A significant positive correlation was noted between livin expression and the TFG score (P<0.05).

CONCLUSIONInhibition of cell apoptosis is more obvious at the ITF of oral squamous cell carcinoma than in the other regions. Livin overexpression at the ITF may indicate greater malignancy and higher likeliness of tumor recurrence and metastasis.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Male ; Middle Aged ; Mouth Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Proteins ; genetics ; metabolism

AIMTo determine how SDF-1 alpha/CXCR4 activates nuclear factor-kappa B (NF-kappaB) and promotes oral squamous cell carcinoma (OSCC) invasion.

METHODOLOGYA lentivirus-based knockdown approach was utilized to deplete gene expression. NF-kappaB activation was evaluated by Western blot analysis and electrophoretic mobility shift (EMSA).

RESULTSWe show that the activation of NF-kappaB by CXCR4 occurs through the Carma3/Bcl10/Malt1 (CBM) complex in OSCC. We found that loss of components of the CBM complex in HNSCC can inhibit SDF-1 alpha induced phosphorylation and degradation of IkappaBalpha, while TNF alpha induced IKK activation remains unchanged. Further, we identified a role for novel and atypical, but not classical, PKCs in activating IKK through CXCR4. Importantly, inhibition of the CBM complex leads to a significant decrease in SDF-1 alpha mediated invasion of OSCC.

CONCLUSIONThe CBM complex plays a critical role in CXCR4-induced NF-kappaB activation in OSCC. Targeting molecular components of the NF-kappaB signaling pathway may provide an important therapeutic opportunity in controlling the progression and metastasis of OSCC mediated by SDF-1 alpha.

Adaptor Proteins, Signal Transducing ; antagonists & inhibitors ; physiology ; B-Cell CLL-Lymphoma 10 Protein ; CARD Signaling Adaptor Proteins ; antagonists & inhibitors ; physiology ; Carcinoma, Squamous Cell ; pathology ; Caspase Inhibitors ; Caspases ; physiology ; Cell Line, Tumor ; Chemokine CXCL12 ; antagonists & inhibitors ; physiology ; Enzyme Activation ; drug effects ; Gene Silencing ; Genetic Vectors ; genetics ; Humans ; I-kappa B Kinase ; drug effects ; I-kappa B Proteins ; metabolism ; Isoenzymes ; antagonists & inhibitors ; Lentivirus ; genetics ; Membrane Proteins ; antagonists & inhibitors ; physiology ; Mouth Neoplasms ; pathology ; Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein ; NF-KappaB Inhibitor alpha ; NF-kappa B ; antagonists & inhibitors ; physiology ; Neoplasm Invasiveness ; Neoplasm Proteins ; antagonists & inhibitors ; physiology ; Phosphorylation ; Plasmids ; genetics ; Protein Kinase C ; antagonists & inhibitors ; Receptors, CXCR4 ; physiology ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo investigate the mRNA expression of matrix metalloproteinase 1 (MMP1) gene in oral squamous cell carcinoma (OSCC) and the paired normal tissues.

METHODSThe differential expression of MMP1 mRNA between 30 OSCC and paired normal tissues were detected with reverse transcription-PCR (RT-PCR).

RESULTSThe relative expression level of MMP1 mRNA in the OSCC tissues showed a 3.26-fold increase in comparison with that in the paired normal tissues (4.06-/+0.52 vs 1.24-/+0.17, P<0.0001). In the 30 OSCC tissues, the relative expression level of MMP1 mRNA was higher in histological grade II/III tissues (4.31-/+0.68) than in grade I (3.87-/+0.57) tissues, higher in OSCC in advanced stages (III/IV) than in tumors in early stages (I/II) (4.18-/+0.67 vs 3.65-/+0.53), and also higher in OSCC with cervical lymph node invasion than in those without cervical lymph node invasion (4.32-/+0.71 vs 3.91-/+0.51), but these differences were not statistically significant (P>0.05).

CONCLUSIONMMP1 gene may play a role in local invasion of OSCC, and can serve as a potential biomarker molecule for diagnosis, treatment and prognostic evaluation of OSCC, with also clinical value for OSCC classification.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; enzymology ; genetics ; pathology ; Female ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Matrix Metalloproteinase 1 ; genetics ; Middle Aged ; Mouth Mucosa ; enzymology ; metabolism ; pathology ; Mouth Neoplasms ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction