Novel cell-surface peptides specific to human oral squamous cell carcinoma using an E. coli peptide display library.

We attempted to find a specific antigen of oral squamous cell carcinoma (SCC) cells that could be safely applied to gene therapy in the conservative clinical treatment of oral cancer. We performed subtraction using normal human keratinocyte cells, followed by selection using four oral SCC cell lines. We isolated three clones from poorly differentiated SCC cells and four from well-differentiated SCC cells. These seven clones adsorbed to the oral SCC cells at rates 10-100 times those of normal human keratinocyte cells. The three clones from the poorly differentiated SCC cells showed the same peptide sequence (LAPRTHP). Of the four clones from the well-differentiated SCC cells, three showed the same peptide sequence (FGTLPGT) and the fourth showed a different one (VTPNSTP). Each peptide sequence may recognize the material that exists specifically on the oral SCC cell cortex. We can expect applications not only for tumor-targeting treatment using a gene therapy virus vector but also for diagnosis using, as a tumor marker, the peculiar SCC surface material that these peptides recognize.

Diagnostic value of dynamic contrast-enhanced MRI in the salivary gland tumors.

To evaluate the diagnostic value of dynamic contrast-enhanced MRI (DCE-MRI) in salivary gland tumors, thirty-five patients (47 lesions) who underwent MR examinations and were histopathologically diagnosed with salivary gland tumors in Okayama University Hospital, between April 1998 and March 2005, were entered in the present study. The parameters included CI(max300) or CI(max600), which was the contrast index (CI) at maximal contrast enhancement upon 300 s or 600 s, and Tmax, which was the time that corresponded to the CI(max300). Washout ratio (WR(300) or WR(600)) was defined as follows: CI(max300)-CI(300s)/CI(max300) or CI(max600)-CI(600s)/CI(max600)x100 (%), where CI(300) or CI(600) was the CI at 300s or 600s after contrast medium administration. We obtained the following results from the analysis of DCE-MRI parameters; (a) The salivary gland tumors were categorized into three CI curve types according to Tmax and WR300; Pleomorphic adenoma; Tmax > 210 s and WR300 < 10%, Warthin tumor; Tmax < 60 s and WR300 > 40%, and malignant tumor; 60s < Tmax < 210 s and 10% < WR300 < 30%; (b) On the basis of the relationship between Tmax and CImax or WR, all pleomorphic adenomas were successfully differentiated from Warthin tumor lesions. Of the 20 pleomorphic adenomas, 18 (90.0%) were successfully differentiated from malignant tumors. All Warthin tumor lesions were successfully differentiated from pleomorphic adenomas and malignant tumors. Of 12 the malignant tumors, 11 (91.7%) were successfully differentiated from pleomorphic adenomas. All malignant tumors were successfully differentiated from Warthin tumors. Thus, DCE-MRI parameters are useful in diagnosing salivary gland tumors on the basis of the combined assessment of Tmax and CImax or WR.

Enhancement effects of test injection with a small amount of MR contrast medium in the oral and maxillofacial region.

PURPOSE: To examine whether the signal intensity of dynamic contrast-enhanced MRI (DCE-MRI) is altered by test injection of 1 ml of contrast medium, and if so, whether this change affects the DCE-MRI analysis. MATERIALS AND METHODS: Six healthy volunteers were examined by DCE-MRI using a Magnevist syringe and/or an Omniscan syringe for the injection of contrast medium. Each scan was performed 10 times using steady-state free precession (3D-FISP), a sequence for DCE-MRI, before and after intravenous injection of 1 ml of the contrast medium. The internal pterygoid muscle, masseter muscle, tongue, parotid gland, submandibular gland, bone marrow of the mandible, subcutaneous adipose tissue, and common carotid artery were determined to be regions of interest (ROI), and the ROI internal average signal intensity was measured. The 10 data sets obtained before or after contrast medium administration for each ROI were evaluated using the paired t-test. RESULTS: The test injection increased the signal intensities of six of eight ROIs, with all 20 experiments in the submandibular gland showing significant differences. There was no significant difference in the two ROIs corresponding to the carotid artery and subcutaneous adipose tissue of the cheek. CONCLUSIONS: The enhanced signal intensity in the tissue might have been caused by the small amount of contrast medium in the test injection. To eliminate this discrepancy caused by the test injection, a pre-contrast scan should be performed when the average signal intensity of an ROI is measured. We therefore believe that the data obtained before a test injection may be important in the analysis of DCE-MRI.

Influence of CpG island methylation status in O6-methylguanine-DNA methyltransferase expression of oral cancer cell lines.

It is known that the O6-methylguanine-DNA methyltransferase (MGMT) gene is susceptible to epigenetic regulation associated with an altered frequency of CpG methylation. To investigate whether epigenetic regulation of the MGMT gene might lead to significant reductions in the expression levels of cancer cells, we sought evidence of a link between the methylation status of the MGMT promoter and the expression levels of seven human oral cancer cell lines. We found two frequently methylated regions: the 5' region extending from nt 690 to nt 893 in the promoter, and the more 3' region extending from nt 1060 to nt 1151 in the untranslated first exon. The 3' region was hypermethylated independently of MGMT expression levels in all cell lines. By contrast, in the three MGMT-downregulated cell lines (SAS, Hep2, HO-1-u-1), the levels of MGMT expression were inversely related to the density of 5' region of the methylated CpGs in the MGMT promoter. Our results implied that the transcriptional inactivation of MGMT might require methylation of the 5' region, but not that of the 3' region in oral cancer cell lines. We further explored the role of methylation in MGMT expression by treating cells with 5-Aza-2'-deoxycytidine (5Aza-dC). 5Aza-dC treatment led to the partial or complete cytosine demethylation of two frequently methylated MGMT regions in all cell lines. 5Aza-dC succeeded in upregulating of the MGMT mRNA levels in only 2 of 7 cell lines (HSC3 and HO-1-u-1), and in fact reduced MGMT mRNA in the other 5 cell lines. Furthermore, 5Aza-dC had an inhibitory effect on MGMT protein levels in all cell lines. Our results suggest that MGMT levels may not revert after 5Aza-dC treatment. Based on our findings, the regulation of MGMT expression appears to be more complex than previously thought, although it is at least partially influenced by CpG methylation. Accordingly, care should be taken interpreting the link between MGMT methylation and expression.

Effects of demethylating agent 5-aza-2(')-deoxycytidine and histone deacetylase inhibitor FR901228 on maspin gene expression in oral cancer cell lines.

Maspin, which belongs to the serine protease inhibitor (serpin) superfamily, has been proposed as a potent tumor suppressor that inhibits cell motility, invasion, angiogenesis, and metastasis. In the present study, we examined the effects of 5-aza-2(')-deoxycytidine (5-aza-dC), a demethylating agent, and FR901228, a histone deacetylase (HDAC) inhibitor, on maspin expression in oral cancer cell lines. The expression levels of maspin mRNA were divided into two groups, which was the maspin low-expressed and high-expressed cell lines in the 12 oral cancer cell lines. The maspin promoter contained only a few methylated CpG sites in the maspin low-expressed cell lines. Moreover, the methylation status was not altered after 5-aza-dC treatment. However, the transcription of the maspin gene was clearly increased following 5-aza-dC treatment in a number of oral cancer cell lines. These results imply that an action of 5-aza-dC is separate from induction of promoter demethylation. Treatment with FR901228 resulted in a time-dependent stimulation of the re-expression of maspin mRNA as early as 4 h after treatment in the maspin downregulated cells. The re-expression of the maspin gene may contribute to the recuperation of biological functions linked to FR901228 such as an inhibitory effect on tumor angiogenesis and cell invasion. These results indicate that maspin and its target genes may be excellent leads for future studies on the potential benefits of FR901228, a HDAC inhibitor, in cancer therapy.

Incidentally found and unexpected tumors discovered by MRI examination for temporomandibular joint arthrosis.

We examined the frequency of incidentally found or unexpected tumors discovered at the time of magnetic resonance imaging (MRI) examinations in the temporomandibular joint (TMJ) region for patients with suspicion of TMJ arthrosis. Five MR images (T1-weighted transverse scout image and proton density and T2-weighted oblique sagittal images at the open and closed mouth) were acquired. In 2776 MRI examinations of TMJ arthrosis, two tumors were discovered. They consisted of an adenoid cystic carcinoma in the deep portion of the parotid gland, and a malignant tumor extending from the infratemporal fossa to the parapharyngeal space. The rate of incidentally founded or unexpected tumors in TMJ examinations was low (0.072%), but the two tumors found were malignant tumors, and therefore, scout image should be carefully examined, not only used for positing the slice.