Xiaoan Tao

Differential gene expression profiles of whole lesions from patients with oral lichen planus

BACKGROUND The molecular biological properties of oral lichen planus (OLP) are largely unknown. The aim of this study was to identify the genes responsible for its pathogenesis at the genome scale using DNA microarray technology. METHODS The RNA samples extracted from the specimens of nine OLP patients and nine controls were analyzed with Affymetrix GeneChip. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis was applied to confirm GeneChip results. RESULTS A total of 985 differentially expressed genes (629 up-regulated and 356 down-regulated) were identified. These genes were involved in many function classifications and biochemical pathways. The results of quantitative RT-PCR analysis of FOXP3, VEGFA, ANGPT1, MMP1, and SCGB2A2 were consistent with their changes demonstrated by GeneChip. CONCLUSIONS This study showed the gene expression profiles of OLP, which were quite distinct from that of healthy controls. These results presented a global view of physiopathologic processes in lesions, which will give important clues to understand pathogenesis and identify new therapeutic targets of OLP.

Expressions of CXCL12/CXCR4 in oral premalignant and malignant lesions.

Objective. The chemokine receptor CXCR4 and its ligand CXCL12 have been suggested to play important roles in the initiation or progression of cancers. The goal of the present study was to investigate alterations of CXCL12/CXCR4 in oral premalignant lesions and oral squamous cell carcinoma (OSCC). Methods. In 13 normal oral epithelia, 24 dysplastic oral leukoplakia (OLK), and 40 OSCC specimens, expressions of CXCL12 and CXCR4 were evaluated by immunohistochemistry. Results. CXCR4 was expressed in 37.5% of OLK and 60% of OSCC. CXCL12 was detected in 50% of OLK and 62.5% of OSCC. In OLK, CXCR4 positive ratio showed no significant difference from normal epithelia, but the CXCL12 positive ratio was significantly higher. Significant relationship between CXCL12 and CXCR4 was found both in OLK and OSCC. Conclusion. Our results indicated that CXCL12/CXCR4 axis may play roles from early steps of oral malignant transformation and contribute to the progress of oral carcinogenesis.

Increased expression of focal adhesion kinase correlates with cellular proliferation and apoptosis during 4-nitroquinoline-1-oxide-induced rat tongue carcinogenesis

BACKGROUND Oral carcinogenesis is a multistep process and requires accumulation and interplay of a series of molecular genetic events. Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that plays an important role in signalling pathways that are initiated at sites of integrin-mediated cell adhesions and by growth factor receptors. Overexpression of FAK has been linked to oral squamous cell carcinoma (OSCC). So, it is hypothesized that FAK expression might contribute to oral carcinogenesis. METHODS During 4-nitroquinoline-1-oxide-induced rat tongue carcinogenesis, FAK protein expression, proliferating cell nuclear antigen (PCNA) and apoptotic nuclei (TUNEL assay) were examined by means of immunohistochemistry. RESULTS Along with the progress of multistage carcinogenesis, FAK expression increased significantly among different histopathological groups with normal mucosa, mild-dysplastic epithelia, moderate-dysplastic epithelia, severe-dysplastic epithelia and in turn OSCC. Furthermore, FAK immunohistochemical index and PCNA-labelling index displayed positive correlation (r = 0.946, P < 0.05), while negative associations were revealed between apoptotic index and final FAK index (r = -0.959, P < 0.05). CONCLUSION Our results implicated a role for FAK in oral carcinogenesis. Inhibition of FAK might be a potential novel treatment strategy in this disease.

Overexpression of miR-155 promotes the proliferation and invasion of oral squamous carcinoma cells by regulating BCL6/cyclin D2

Although microRNA-155 (miR-155) is known to play an important role in many cancers, its expression and function in oral squamous cell carcinoma (OSCC) was not fully understood. Thus, in the present study, we investigated the expression of miR-155 and also the role this miR plays in OSCC. We used the OSCC cell line (CAL27) and paired tumor and non-tumor tissue samples from patients with OSCC in order to detect the expression of miR-155. Cell proliferation, migration and invasion assays were then undertaken in order to determine the effect of miR-155 on the biological behavior of CAL27 cells following transient transfection with miR-155 mimic and antagomir. The regulatory effect of miR-155 on its target gene B-cell CLL/lymphoma 6 (BCL6) and downstream gene cyclin D2 (CCND2) was also analyzed. We found that miR-155 expression in OSCC cell and tumor tissues was significantly higher than that of the controls. We noted that the miR-155 mimic enhanced CAL27 cell proliferation, migration and invasion ability, downregulated BCL6 levels, and increased cyclin D2 expression. However, we noted that abrogating miR-155 with the miR-155 antagomir suppressed CAL27 cell proliferation, migration and invasion, upregulated BCL6 and reduced cyclin D2 expression. These results indicate that miR-155 plays a tumor-promoting role in OSCC by regulating the BCL6/cyclin D2 axis.

Chemokine (CC motif) ligand 18 upregulates Slug expression to promote stem‐cell like features by activating the mammalian target of rapamycin pathway in oral squamous cell carcinoma

Chemokine (CC motif) ligand 18 (CCL18) is involved in remodeling of the tumor microenvironment and plays critical roles in oncogenesis, invasiveness, and metastasis. We previously investigated the overexpression of CCL18 in primary oral squamous cell carcinoma (OSCC) tissues and its association with advanced clinical stage in OSCC patients. However, the underlying mechanisms of this CCL18‐derived activity remains unidentified. This study showed exogenous CCL18 increased cell migration and invasion and induced cell epithelial–mesenchymal transition (EMT), and that E‐cadherin, an epithelial marker, decreased and N‐cadherin, a mesenchymal marker, increased, compared to negative control in OSCC cells. Furthermore, we detected that CCL18 induced the acquisition of cancer stem(‐like) cell characteristics in oral cancer cells, but also found a significantly positive correlation between the expression of CCL18 and Bmi‐1 (P < 0.001) in OSCC surgical specimens by immunohistochemistry analysis. The expression of octamer‐binding transcription factor 4 and Bmi‐1 were significantly upregulated, and proportions of aldehyde dehydrogenasehigh+ cells and CD133+ cells were markedly increased in CCL18‐treated cells compared to untreated cells. Sphere formation ability was observably enhanced when cells were continually exposed to high levels of CCL18. Moreover, CCL18 upregulated Slug expression by stimulating the mammalian target of rapamycin (mTOR) signaling pathway in OSCC cell lines. Inhibition of the mTOR pathway by INK128, or Slug knockdown by RNA interference, reversed CCL18‐induced EMT and the stemness response at both molecular and functional levels. In conclusion, our data suggested that CCL18 upregulated Slug expression to promote EMT and stem cell‐like features by activating the mTOR pathway in oral cancer. These findings provide new potential targets for the early diagnosis and treatment of OSCC.

Genome-wide analysis reveals the active roles of keratinocytes in oral mucosal adaptive immune response.

To elucidate the roles of oral keratinocytes in the adaptive immune response of oral mucosa, global gene expression analysis was performed by microarray technique and integrating computational methods, including hierarchical clustering, biological process Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes pathway analysis, self-organizing maps (SOMs) and biological association network analysis (BAN). Raw data from microarray experiments were uploaded to the Gene Expression Omnibus Database, http://www.ncbi.nlm.nih.gov/geo/ (GEO accession GSE28035). We identified 666 differentially expressed genes in the early stage (48 h) and 993 in the late stage (96 h) of the oral mucosal adaptive immune response. The analysis revealed that oral keratinocytes exerted diverse biological functions in different stages of immune response. Specifically, in 48 h the differentially expressed genes encompassed an array of biological ontology associated with immune response, such as antigen processing and presentation, and positive regulation of T-cell-mediated cytotoxicity. Several pathways which have been reported to be critical in inflammation, including mitogen-activated protein kinase pathway, were activated. Furthermore, after BAN construction, some putative hub genes and networks such as interleukin-1α and its subnetwork were recognized. Taken together, these results give substantial evidence to support the active roles of keratinocytes in the oral mucosal adaptive immune response.

Distribution and quantity of label‐retaining cells in rat oral epithelia

BACKGROUND Label-retaining cells (LRCs), the presumptive stem cells, have been detected in the mouse and hamster oral epithelia, but the data on LRCs in rat oral epithelia have not been available yet. The aim of this study was to identify LRCs in oral squamous epithelia of rat. METHODS Fifty-four two-week-old Sprague-Dawley rats were injected intraperitoneally with BrdU twice daily for four consecutive days. The BrdU-labeled rats were sacrificed at 2 h, week 2, 4, 6, 8, 10, 12, 14 and 16 after the last BrdU injection. The tissues of cheek, tongue and palate were analyzed by techniques of fluorescence-activated cell sorting (FACS) and immunohistochemistry. RESULTS The number of BrdU-labeled cells quantified by FACS increased within 2 weeks after labeling, then, decreased gradually until week 10. After week 10, the BrdU-labeled cells were found to locate mainly in basal layer and their number kept consistent at 3-7% of total oral epithelial cells. The number of BrdU-labeled epithelial cells in palate was statistically higher than that in cheek or tongue at the same time point. CONCLUSIONS This study showed that the number and the distribution of BrdU-labeled epithelial cells stabilized from 10 weeks after labeling. Therefore, these BrdU-labeled cells after a 10-week chase were considered oral epithelial LRCs.

The positive correlation of the CCL2-CCR2 axis with the disease activity may indicate the fundamental role in the pathogenesis of oral lichen planus

BACKGROUND The important roles of CCL2 and its receptor CCR2 had been reported in a series of inflammatory disorders. However, few studies investigated the potential role of CCL2/CCR2 axis in oral lichen planus (OLP). Therefore, this study aimed to detect the expression of CCL2 and CCR2 in OLP lesions and compare their changes before and after treatment. METHODS CCL2 and CCR2 expression was investigated using immunohistochemical staining and real-time RT-PCR in 32 patients with OLP and eight controls. Moreover, changes in their expression after treatment with triamcinolone acetonide were assessed in lesions from three patients. RESULTS CCL2+ and CCR2+ cells were few in the controls and remarkably increased in the epithelial and subepithelial layers of lesions (n = 32, all P < 0.001). However, the densities of CCL2+ and CCR2+ cells were not significantly different between reticular (n = 12) and erythematous/erosive lesions (n = 20), although they significantly decreased after treatment (627.7 ± 108.2 vs. 258.3 ± 148.3, P = 0.017; 1034.7 ± 74.6 vs. 648 ± 77.6, P = 0.003, respectively). CCL2+/CCR2+ cell numbers were positively correlated with disease activity (correlation coefficient, 0.588; P < 0.001; correlation coefficient, 0.409; P = 0.02, respectively). CONCLUSIONS The results of this study indicated that the CCL2-CCR2 axis was involved in the pathogenesis of OLP and was positively correlated with disease activity.

The dynamic changes of circulating OCN cells versus insulinlike growth factor-I during primary healing of orthognathic surgeries

OBJECTIVE The objective of this study was to determine the dynamic changes of circulating osteocalcin(+) (OCN(+)) cells and insulinlike growth factor-I (IGF-I) in peripheral blood during early primary repair of jaw bones in patients with orthognathic surgery. STUDY DESIGN The expression of bone-related genes was detected by RT-PCR in circulating OCN(+) cells. The numbers of OCN(+) cells and serum level of IGF-I were determined by flow cytometry, immunocytochemical staining, and ELISA. RESULTS OCN(+) cells significantly increased in peripheral blood, and reached the peak at 1 to 2 weeks after surgery (P < .05). IGF-I in patients significantly decreased 1 week after surgery (P < .05), and then returned gradually to the normal level. There was no significant correlation between the number of circulating OCN(+) cells and the level of IGF-I (P > .05). CONCLUSIONS These findings suggested that circulating OCN(+) cells, at least in part, could be mobilized in response to bone injury, and contribute to bone repair in patients with orthognathic surgery.