Accuracy of Mandibular Reconstruction with a Vascularised Iliac Flap Using 3D Templates: a Systematic Review.

OBJECTIVE: To conduct a systemic review for guidance regarding the application of templates in mandibular reconstruction with vascularised iliac flaps. METHODS: By searching PubMed, EMBASE and the Cochrane Library and collecting relevant literature, information about the types and accuracy of templates was extracted. Data relating to surgical time were also included for further analysis. RESULTS: Eight studies were included. The data analysis showed that the accuracy of operations with templates was higher than that of conventional surgery. The mean deviation was between 0.70 and 3.72 mm. The operational time was shortened to 314.4 minutes and the graft ischemic time was reduced to 15.6 to 26.8 minutes. Application of functional or specifically designed templates can improve the accuracy and shorten surgical time. CONCLUSION: Templates can increase the accuracy and efficiency of mandibular reconstruction with vascularised iliac flaps, which will benefit patients' prognosis and subsequent functional restoration. Further studies should be conducted into application of templates to improve the accuracy of reconstructions.

Association between interleukin-10 gene polymorphisms and risk of oral carcinoma: A meta-analysis.

BACKGROUND: The single nucleotide polymorphisms (SNPs) of Interleukin-10 (IL-10) gene have been linked with the risk of oral carcinoma (OC) in a relatively small sample size. Our study aims to investigate the pooled associations by conducting a meta-analysis of published studies. METHODS: PubMed, Web of Science and Google Scholar databases were searched to identify eligible studies published in English before October 2019. The odds ratio (OR) with a 95% confidence interval (CI) was used to assess association. The publication bias was detected by Begg's test. Sensitivity and cumulative analyses were performed to evaluate the stability of crude results. RESULTS: The meta-analysis involved eight studies. Significant associations were certified between IL-10 gene -1082A/G polymorphism and susceptibility of OC for A vs. G (OR=1.817, 95% CI: 1.481-2.230), AA vs. GG (OR=3.436, 95% CI: 2.281-5.175), dominant genetic model (OR=2.913, 95% CI: 1.939-4.376), and recessive genetic model (OR=1.886, 95% CI: 1.372-2.594) in overall population, East Asians and South Asians. In addition, the significant association between -592A/C polymorphism of the gene and susceptibility of OC were detected in South Asians. CONCLUSIONS: The meta-analysis results support that the IL-10 gene -1082G allele is a risk factor for OC in East Asians and South Asians, and IL-10 gene -592C allele is a protective factor for the disease.

Sonic Hedgehog Signalling Activation Contributes to ALCAM Over-Expression and Poor Clinical Outcome in Patients with Oral Squamous Cell Carcinoma.

OBJECTIVE: To investigate the Sonic Hedgehog (SHH) signalling molecules and activated leukocyte cell adhesion molecule (ALCAM) expression in the mechanisms regulating invasion and metastasis in oral squamous cell carcinoma (OSCC). METHODS: The expressions of SHH signalling molecules Gli family zinc finger 1/2 (Gli1/Gli2), as well as ALCAM expression, was analysed in 101 OSCC patients by immunohistochemistry. The potential relationship between Gli1/Gli2 and ALCAM in regard to invasion and metastasis were studied by western blot, invasion and wound-healing assays. RESULTS: Gli1, Gli2 and ALCAM were expressed in 54.5%, 49.5% and 47.5% of the 101 OSCC specimens, respectively. High expression of ALCAM was associated with shorter survival in the patient population (P = 0.018), which was independent of other clinical parameters. Notably, when both ALCAM expression and positive nodal status were considered, an enhanced prediction of clinical outcomes was achieved (P = 0.001). In OSCC cell lines, down-regulation of ALCAM resulted in reduced cell invasion and metastasis. Importantly, SHH activation increased the half-life of ALCAM leading to ALCAM accumulation and increased cell invasion and migration. CONCLUSION: ALCAM over-expression in OSCC is an independent prognostic factor for OSCC patients. Its over-expression may be the result of the activation of the SHH signalling pathway and contributes to OSCC progression.

A three-lncRNA signature derived from the Atlas of ncRNA in cancer (TANRIC) database predicts the survival of patients with head and neck squamous cell carcinoma.

OBJECTIVE: Long non-coding RNAs (lncRNAs) have important biological functions and can be used as prognostic biomarkers in cancer. To identify a lncRNA prognostic signature for head and neck squamous cell carcinoma (HNSCC). METHOD: We analysed RNA-seq data derived from the TANRIC database to identify a lncRNA prognostic signature model using the orthogonal partial least squares discrimination analysis (OPLS-DA) and 1.5-fold expression change criterion methods. The prognosis prediction model based on the lncRNA signatures and clinical parameters were evaluated using the 5-fold cross validation method. RESULTS: A total of 84 out of 3199 lncRNAs were significantly associated with the survival of patients with HNSCC (log-rank test P<0.01). Using the OPLS-DA and 1.5-fold change selection criterion, 5 lncRNAs (KTN1-AS1, LINC00460, GUSBP11, LINC00923 and RP5-894A10.6) were further selected. The prediction power of each combination of the 5 lncRNAs was evaluated through the receiver operating characteristic (ROC) curve and a three-lncRNA panel (KTN1-AS1, LINC00460 and RP5-894A10.6) achieved the highest prognostic prediction power (AUC 0.68, 95% CI 0.60-0.76, P<0.0001) in the cohort. The patients were categorized into high- and low-risk groups based on their three-lncRNA profiles. Patients with high-risk scores had worse overall survival than those with low risk scores in the cohort (log-rank test P=0.0003). Multivariable Cox regression analyses showed that the lncRNA signature and tumour grade were independent prognostic factors for patients with HNSCC. CONCLUSIONS: Our findings showed that the three-lncRNA signature might be a novel biomarker for the accurate prognosis prediction of patients with HNSCC.

CD73 is associated with poor prognosis in HNSCC.

CD73 is a cell surface immunosuppressive enzyme involved in tumor progression and metastasis. While patients whose cancer cells express elevated CD73 are typically associated with an unfavorable outcome, the clinical impact of CD73 expression in patients with Head and neck squamous cell carcinoma (HNSCC) remains unclear. In the present study, we investigated the prognostic significance of CD73 in HNSCC using gene and protein expression analyses. Our results demonstrate that high levels of CD73 are significantly associated with reduced overall survival in patients with HNSCC. We also investigated the functional role of CD73 in vitro and demonstrated that CD73 promotes HNSCC migration and invasion through adenosine A3R stimulation and the activation of EGF/EGFR signaling. Moreover, in vivo xenograft studies demonstrated that CD73 promotes tumorigenesis. In conclusion, our study highlights a role for CD73 as a poor prognostic marker of patient survival and also as a candidate therapeutic target in HNSCCs.

Tricholemmal carcinoma of the head and neck region: A report of 15 cases.

Tricholemmal carcinoma is an extremely rare malignancy of the skin, and its biological behavior and management is controversial. The objective of the present study was to investigate the clinicopathological characteristics and management of tricholemmal carcinoma of the head and neck region. The study analyzed 15 patients with tricholemmal carcinoma. Demographic and clinical data were collected, and features associated with the management and prognosis of tricholemmal carcinoma were analyzed. Two of the 15 patients were lost to follow-up. The results showed that, during the follow-up period, 5 of the 13 available patients succumbed to the causes of recurrence (n=3), neck lymph node metastasis (n=1) and Parkinson's disease (n=1). No patients developed distant metastasis. The disease-free survival (DFS) and overall survival (OS) were 31.1±7.8 and 32.9±7.4 months (mean ± SE), respectively, and the DFS and OS rates were 69.2 and 61.5%, respectively. In conclusion, the biological behavior of tricholemmal carcinoma is locoregionally aggressive. The recommended management for head and neck tricholemmal carcinoma is radical resection and neck dissection, and post-operative radiotherapy may be considered for high-risk patients.

[Screening of the gene mutation in D-loop region of mitochondrial DNA in oral squamous cell carcinoma].

OBJECTIVE: To investigate the gene mutation in D-loop region of mitochondrial DNA (mtDNA) in oral squamous cell carcinoma (OSCC) tissue and to explore the role of the gene mutation in D-loop region in the OSCC tumorigenesis. METHODS: mtDNA was obtained from cancer, paracancerous and normal mucosa tissues of thirty patients with OSCC. The D-loop regions of mtDNA were amplified with PCR, sequencing and then analyzed by Chromas software and BLAST to identify the mutation site. RESULTS: Mutation in the D-loop region was found in eight cases, with the mutation rate of 27%. There were nine mutations totally, including one point mutation, two base deletions, three insertion mutations, three heterozygous mutations. In these mutations, base deletions were different from each other and heterozygous mutations had no same mutation form, while the three insertion mutations were same, the insertion of base C. One case had T/A heterozygous mutation and base C insertion at the same time. CONCLUSIONS: There were mutations in mtDNA D-loop in OSCC, but the relationship between occurrence of OSCC and mutation of mtDNA needs further study.

Repression of G protein-coupled receptor family C group 5 member A is associated with pathologic differentiation grade of oral squamous cell carcinoma.

BACKGROUND: G protein-coupled receptor family C group 5 member A (GPRC5A), a member of G protein-coupled receptor family, has been shown to function as a tumor suppressor in lung tissue. The biological functions of GPRC5A have therefore been linked to lung tissue. However, the biological significance of this gene product remains obscure. In this study, we investigated the expression of GPRC5A proteins in normal oral tissue and oral squamous cell carcinoma (OSCC), and we characterized its biological activity in OSCC cell lines. METHODS: Western blot analysis and immunohistochemical staining were used to investigate the expression of GPRC5A in both OSCC cell lines and clinical samples. GPRC5A stable transfectants and their parental OSCC cells were characterized for their biological activities in anchorage-independent growth. RESULTS: High levels of immunohistochemical GPRC5A expression were detected in normal oral tissue, especially differentiated area. In contrast, GPRC5A expression was dramatically repressed in OSCCs (P < 0.01). The immunohistochemical GPRC5A expression was moderately well differentiated, but greatly repressed in moderately differentiated OSCCs and completely repressed in poorly differentiated OSCCs. Overexpression of GPRC5A in OSCC CAL27 cells resulted in a suppressed anchorage-independent growth activity, a transforming phenotype. CONCLUSIONS: GPRC5A is expressed in normal oral epithelium. Repression of GPRC5A is associated with poorly differential grade of OSCCs. Overexpression of GPRC5A in OSCC cell line reversed the malignant phenotype. Thus, GPRC5A is important for homeostasis in oral tissue, and deletion or repression of this gene may involve in tumorigenesis of OSCCs and may serve as a prognostic marker for malignant type of OSCCs.

Single nucleotide polymorphisms in the u-PA gene are related to susceptibility to oral tongue squamous cell carcinoma in the Northern Chinese Han population.

AIM: The purpose of this study was to determine whether susceptibility to oral tongue squamous cell carcinoma (OSCC) is related to polymorphisms in the u-PA gene. METHODS: We examined the rs2227564 C/T and rs2227562 G/A single nucleotide polymorphisms (SNPs) in 196 OSCC patients and 201 age- and gender- matched controls via direct sequencing and PCR-RFLP methods. RESULTS: Significant differences were found in allelic and genotypic distributions of the rs2227564 and rs2227562 loci when comparing cases and controls. In addition, logistic analyses indicated that the rs2227564 C/T genotype was related to a 1.52-fold increased risk of developing OSCC (adjusted OR=1.521, 95%CI: 1.144~2.022, P=0.004). Linkage disequilibrium analysis was conducted and no association between the two loci was found (D'=0.031, r2=0.000). CONCLUSIONS: Our findings provide evidence that the rs2227564 C/T SNP in the u-PA gene is associated with the development of OSCC.

Expression of podoplanin in salivary gland adenoid cystic carcinoma and its association with distant metastasis and clinical outcomes.

Distant metastasis is a common cause of mortality in patients with salivary gland adenoid cystic carcinoma (SACC). However, presently, the development of distant metastasis is unable to be predicted in clinical practice. Recent studies have shown that overexpression of podoplanin is associated with metastasis and survival in patients with several cancer types. The purpose of the present study was to determine whether podoplanin is overexpressed in SACC and whether such overexpression is associated with distant metastasis and survival. Podoplanin expression was determined using immunohistochemistry (IHC) in tumors from 40 SACC patients. The expression status was analyzed in regards to patient clinicopathological parameters and survival rates. Overexpression of podoplanin was detected in 13 (32.5%) of the 40 tumors. Overexpression was significantly associated with disease-free survival (P=0.025) and distant metastasis (P=0.015), although it was not associated with recurrence and overall survival. In conclusion, podoplanin is overexpressed in a subset of SACCs and may be a biomarker predicting distant metastasis in patients with SACC.

XIAP is a predictor of cisplatin-based chemotherapy response and prognosis for patients with advanced head and neck cancer.

BACKGROUND: Approximately 60-80% of patients with advanced head and neck squamous cell carcinoma (HNSCC) die within five years after diagnosis. Cisplatin-based chemotherapy is the most commonly used palliative treatment for these patients. To evaluate the prognostic value of X-linked inhibitor of apoptosis (XIAP) level as a potential biomarker in these patients, we investigated the relationship between XIAP expression and cisplatin response of these patients and their prognosis. METHODOLOGY/PRINCIPAL FINDINGS: Sixty patients with advanced HNSCC were recruited in this study. Expression of XIAP was examined both before and after chemotherapy and was correlated with chemotherapy response, clinicopathology parameters and clinical outcomes of the patients. We found that XIAP was expressed in 17 (20.83%) of the 60 advanced HNSCC samples and the expression was significantly associated with cisplatin resistance (P = 0.036) and poor clinical outcome (P = 0.025). Cisplatin-based chemotherapy induced XIAP expression in those post-chemotherapy samples (P = 0.011), was further associated with poorer clinical outcome (P = 0.029). Multivariate analysis demonstrated that only alcohol consumption, lymph node metastasis and XIAP level were independently associated with the prognosis of advanced HNSCC patients. Inhibiting XIAP expression with siRNA in XIAP overexpressed HNSCC cells remarkably increased their sensitivity to cisplatin treatment to nearly a 3 fold difference. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that XIAP overexpression plays an important role in the disease course and cisplatin-resistance of advanced HNSCC. XIAP is a valuable predictor of cisplatin-response and prognosis for patients with advanced head and neck cancer. Down-regulation of XIAP might be a promising adjuvant therapy for those patients of advanced HNSCC.

[Investigation of mRNA expression of collagen genes in oral squamous cell carcinoma and paired normal tissue].

OBJECTIVE: To investigate the mRNA expression of collagen genes in oral squamous cell carcinoma (OSCC) and paired normal oral mucosal tissue. METHODS: The differential mRNA expressions of collagen genes between 30 OSCC tissues and the paired normal oral mucosal tissues were detected by RT-PCR. RESULT: The relative expression level of COL1A1 mRNA in the 30 cancerous tissues was up-regulated by 2.78 folds as compared with its expression in the paired normal samples, suggesting its significant overexpression in OSCC (P<0.001). The expression levels of COL1A2, COL4A1, COL4A2, and COL5A2 mRNA in the cancerous tissues were up-regulated by 1.07, 1.15, 1.27, and 1.16 folds compared to those in paired normal samples (P>0.05). CONCLUSION: COL1A1 mRNA overexpression may play an important role in the carcinogenesis of OSCC and can be a potential molecular marker of OSCC.

The expression of EMP1 is downregulated in oral squamous cell carcinoma and possibly associated with tumour metastasis.

AIMS: To investigate the expression of EMP1 in oral squamous cell carcinoma (OSCC) tissues and its correlation with available clinical parameters of patients with OSCC. METHODS: The mRNA levels of EMP1 were measured in 18 paired OSCC and corresponding adjacent normal tissues using RT-PCR. Another 45 pairs of OSCC samples were selected to detect the mRNA level of EMP1 using quantitative RT-PCR (qRT-PCR). The protein levels of EMP1 were also evaluated in 60 cases of patients with OSCC using immunohistochemical staining. The correlation between EMP1 expression and clinical parameters was analysed with non-parametric analysis. RESULTS: The results of RT-PCR and qRT-PCR showed that, compared with the paired normal tissues, the mRNA levels of EMP1 were significantly decreased in OSCC. The immunohistochemical results indicated that the EMP1 protein was also downregulated in OSCC (p=0.031). Decreased expression of EMP1 was significantly correlated with clinical stage (p=0.002) and lymph node metastasis (p=0.044) of patients with OSCC. Meanwhile, there was a significant difference between OSCCs at early and advanced stages (p=0.003), and between OSCCs with lymph node metastasis and no lymph node metastasis (p=0.045), respectively. CONCLUSIONS: The results suggest that EMP1 may be a tumour suppressor and associated with lymph node metastasis in OSCC.

Epigenetic silencing of MAL, a putative tumor suppressor gene, can contribute to human epithelium cell carcinoma.

BACKGROUND: To identify new and useful candidate biomarkers in head and neck squamous cell carcinoma (HNSCC), we performed a genome-wide survey and found that Myelin and lymphocyte-associated protein (MAL) was a gene that was markedly down-regulated in HNSCC. Hence, we investigated the mechanism of MAL silencing and the effects of MAL on the proliferation, invasion, and apoptotic potential in HNSCC. RESULTS: MAL was significantly down-regulated in 91.7% of HNSCC specimens at the mRNA level as compared with adjacent normal tissues (P = 0.0004). Moreover, the relative transcript levels of the MAL gene were remarkably decreased by five-fold in nine HNSCC cell lines as compared with normal head and neck epithelium cells. MAL gene expression was restored in 44%, 67%, and 89% in HNSCC cell lines treated with TSA, 5-Aza-dC, and TSA plus 5-Aza-dC, respectively. Furthermore, bisulfate-treated DNA sequencing demonstrated that the two CpG islands (that is, M1 and M2) located in MAL promoter region were completely methylated in the HNSCC cell lines (CpG methylated ratio was more than 90%), and only one CpG island (that is, M1) was partially methylated in HNSCC tissues (CpG methylated ratio between 20% and 90%). A significant reduction in cell proliferation and a change in the cell cycle profile were also observed in MAL transfectants. Matrigel assay demonstrated that the invasiveness of HNSCC cells significantly decreased. A significant increase in the population of apoptotic cells was observed in MAL transfected cells. The exogenous expression of the MAL gene suppressed malignant phenotypes, while the cell death induced by MAL gene transfer was a result of apoptosis as demonstrated by the induction of cleavage of the poly (that is, ADP-ribose) polymerase. Additionally, tumor growth was suppressed in cells expressing MAL as compared with cells not expressing MAL. CONCLUSION: Our data suggest that the epigenetic inactivation of MAL, as a candidate tumor suppressor gene, can contribute to human epithelial cell carcinoma and may be served as a biomarker in HNSCC.

Correlation of NF-kappaB signal pathway with tumor metastasis of human head and neck squamous cell carcinoma.

BACKGROUND: Nuclear factor-kappa B (NF-kappaB) signaling constitutes a key event in the multistep process of carcinogenesis, progression and treatment in many cancer types. However, the significance of NF-kappaB pathway for complex and tissue-specific aspects of head and neck cancer progression, such as invasion and metastasis, is less understood. METHODS: The expression of NF-kappaB p65 in squamous cell carcinoma of the head and neck (SCCHN) clinical specimens by immunohistochemistry. The role of NF-kappaB activity in head and neck squamous cell carcinoma was determined by western blot, reporter assay and EMSA analysis in vitro and metastasis assays in vivo in different metastatic potential tumor cells. Furthermore, the apoptosis rate and expression of metastasis-related protein such as MMP9 and VEGF were examined by Annexin V/PI staining and Western blot, respectively. RESULTS: A higher level of active nuclear-localized NF-kappaB was observed in the metastatic SCCHN specimens group (p < 0.01). The NF-kappaB activities of SCCHN cell lines with different metastatic potentials were then determined and in excellent agreement with results found in SCCHN specimens, highly metastatic SCCHN cell lines expressed high level of NF-kappaB activity. The treatment of highly metastatic SCCHN cells with NF-kappaB inhibitors reduced the in vitro cell invasion capacity of the cells without affecting the apoptotic rate. Additionally, the NF-kappaB inhibitors significantly inhibited the experimental lung metastasis of Tb cells and lymph node metastasis of TL cells in nude mice. Furthermore, the expression of metastasis-related proteins, such as matrix metalloproteinase 9 and vascular endothelial growth factor, was inhibited by pyrrolidine dithiocarbonate. CONCLUSIONS: This study suggests that NF-kappaB activity significantly contributes to tumor hematologic and lymphatic metastases and may aid in the development of early detection methods or therapies targeting non-conventional molecular targets.

[Effect of Matrine on cell cycle and human telomerase reverse transcriptase of human salivary adenoid cystic carcinoma].

OBJECTIVE: To investigate the effect of the traditional Chinese medicine Matrine on cell cycle and human telomerase reverse transcriptase (hTERT) of human ACC-M cell lines. METHODS: Different concentrations of Matrine were used in the medium of ACC-M cells. Change of cell cycle were detected by flow cytometry after ACC-M cell were cultivated with different concentrations Matrine (0.25, 0.50, 0.75, 1.00 mg x mL(-1)). Expression of hTERT was investigated by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescene and flow cytometry quantitative analysis. RESULTS: Matrine caused obviously the GdG1 phase block and inhibited proliferation of ACC-M cells. At same time, this effect was positive correlation to Matrine concentration and treat time. Matrine can inhibit the expression of hTERT mRNA and protein. CONCLUSION: Matrine can obviously inhibit cell cycle and down-regulate expression of hTERT. Inhibition of cell cycle is possible correlation with down-regulation expression of hTERT.

[Inflammatory factors promote oral squamous cell carcinoma cells metastasis, via nuclear factor kappa B signal pathway in vitro].

OBJECTIVE: To investigate the roles of inflammatory factors and nuclear factor kappa B (NF-kappaB) signal pathway in metastasis of oral squamous cell carcinoma. METHODS: The oral squamous cell carcinoma cell lines with highly metastasis potential (Tb) and lower metastasis potential (Tca8113) were used in this study. The levels of NF-kappaB activity in oral squamous cell carcinoma cell lines were determined by Western blotting and luciferase reporter assay. pBalphabe-IkappaBalpha-SR expression vector or NF-kappaB inhibitor pyrolidinedithiocarbamate (PDTC) was used to inhibit NF-kappaB, and cell migration was examined by transwell assay. The secretion of tumor necrosis factor-alpha (TNF-alpha), IL-1alpha, IL-6, IL-8 and GM-CSF proinflammatory cytokines was determined by ELISA when Tb cells were transfected with pBalphabe-SR-IkappaBalpha or treated with PDTC. RESULTS: Western blotting showed that the levels of phosphorIkappaBalpha and phosphor-p65 were highly expressed in Tb cells. Tb cells had high level of constitutive NF-kappaB activity and were more sensitive to TNF-alpha. The migration of highly metastatic Tb cells, either transfected with dominant-negative mutant inhibitor pBalphabe-SR-IkappaBalpha or treated with PDTC, was suppressed when determined by transwell assay. The secretion of proinflammatory cytokines, including TNF-alpha, IL-1alpha, IL-6, IL-8 and granulocyte-macrophage colony stimulating factor (GM-CSF), was inhibited by pBalphabe-SR-IkappaBalpha transfection or PDTC treatment. CONCLUSIONS: The inflammatory factors such as TNF-alpha could promote oral squamous cell carcinoma cell metastasis via NF-kappaB signal pathway.

[Effects of Podoplanin on cell proliferation and cell cycle in oral leukoplakia cells].

OBJECTIVE: To investigate the effects of Podoplanin on cell proliferation and cell cycle in oral leukoplakia cells. METHODS: Oral leukoplakia cell line (Leuk-1) transfected with pCMV-Podoplanin (A4-1) or pCMV (B4-1) was used in this study. The levels of mRNA and protein of Podoplanin were detected by real-time PCR and Western boltting in A4-1, B4-1 and Leuk-1 cells. The localization of Podoplanin was detected by confocal microscope. Cell growth and proliferation was examined by methyl thiazolyl tetrazolium (MTT) assay and cell cycle was detected by flow cytometry (FCM). RESULTS: The value of Podoplanin mRNA level in A4-1 cells was 0.022, which was significantly higher than the values in B4-1 (0.001) and Leuk-1 cells (0.002), P < 0.05. The gray scale of Podoplanin protein in A4-1 cells was significantly higher than in the control groups (P < 0.05). The expression of Podoplanin was observed in cell plasm and membrane of A4-1, B4-1 and Leuk-1 cells. But the expression level of Podoplanin in A4-1 cells was higher than in control groups. A4-1 cells grew faster than Leuk-1 cells. The proliferation rate after 3 days of culture in A4-1 cells was 40.4% higher than that in B4-1 cells (P < 0.05). The G2-M phase (24.89 +/- 4.55)% and PI (0.57 +/- 0.06) of A4-1 cells were significantly higher than that in B4-1 cells [(4.13 +/- 5.24)%, (0.41 +/- 0.04)] and Leuk-1 cells [(4.69 +/- 7.42)%, (0.40 +/- 0.02)], P < 0.05. CONCLUSIONS: Over expression of Podoplanin accelerated the growth and proliferation of oral leukoplakia cells. Podoplanin may be one of key genes in the malignant transformation of oral leukoplakia.

[Role of NF-kappaB pathway in shikonin induced apoptosis in oral squamous cell carcinoma Tca-8113 cells].

PURPOSE: To investigate the role of NF-kappaB signal transduction pathway in apoptosis induced by shikonin in human tongue squamous cell carcinoma Tca-8113 cell line. METHODS: Expression of IkappaBa, phosphatase-IkappaBa, bcl-2 and Bax proteins were detected by Western blot, NF-kappaB DNA-binding activity was detected by electrophoretic mobility shift analysis (EMSA), and activities of caspase 3, caspase 8 and caspase 9 were analyzed by enzyme linked immunosorbent assay(ELISA). The data was analyzed by one-way ANOVA test and t test using SPSS12.0 software package. RESULTS: The expression of phosphatase-IkappaBa protein and the nuclear NF-kappaB DNA-binding activity was significantly decreased in shikonin treated cells by Western and EMSA. Bcl-2 protein expression was also decreased in the process. The activity of all the three proteases was elevated and pancaspase inhibitor Z-Asp-CH2-DCB could protect Tca8113 cells from shikonin-induced apoptosis(P=0.02). CONCLUSIONS: Anti-tumor effects of shikonin in Tca-8113 cells act at least partially through the inactivation of NF-kappaB pathway and subsequent activation of protease caspase family. Pharmacologic inhibition of the NF-kappaB activity by shikonin might be a powerful treatment option for OSCC.

MicroRNAs contribute to the chemoresistance of cisplatin in tongue squamous cell carcinoma lines.

MicroRNAs (miRNAs) are small non-coding RNAs that function as negative regulators of gene expression. They are strongly implicated in human cancers, including oral squamous cell carcinoma (OSCC). Evidence for the involvement of miRNAs as important regulators of chemosensitivity and chemoresistance in OSCC is not well understood. In this study, miRNA microarray was firstly used to compare the differential miRNAs levels between the cisplatin-sensitive tongue squamous cell carcinoma line (Tca8113) and its cisplatin-resistant subline (Tca/cisplatin). Three miRNAs of miR-21, -214, and -23a were validated by miRNAs real-time PCR, and intervened by anti-miRNA oligonucleotides (miR-214 and -23a) and pre-miRNA plasmid transfection (miR-21). Further relationship between miR-23a and DNA topoisomerase II beta (TOP2B) on the chemoresistance against cisplatin was studied. There were 19 out of 480 differential miRNAs between the Tca8113 and Tca/cisplatin cells. miR-214 and -23a were found increased as with chemoresistance against cisplatin in the Tca/cisplatin cells while miR-21 was found decreased as with chemosensitivity for cisplatin in the Tca/cisplatin cells. Intervention of these three miRNAs could decrease the chemoresistance against cisplatin in Tca/cisplatin cells. Transfection of anti-miR-23a into the Tca/cisplatin cells could increase the TOP2B protein expression. Our results suggest the existence of differential miRNAs with chemosensitivity and chemoresistance between the cisplatin-sensitive and resistant tongue squamous cell carcinoma lines. miR-21 serves as a chemosensitive miRNA, while miR-214 and -23a serve as chemoresistant miRNAs in the tongue squamous cell carcinoma lines. miR-23a is an up-stream regulator of TOP2B to realize the chemoresistance of cisplatin.