Usefulness of contrast enhanced-MRI in the diagnosis of unicystic ameloblastoma.

The radiographic features of unicystic ameloblastoma (UA) are typically unilocular and round area of radiolucency. Therefore, this type of lesion is often misdiagnosed as an odontogenic keratocyst or a dentigerous cyst. UA should be differentiated from odontogenic cysts because the former have a higher rate of recurrence than the latter. In the present study, we performed contrast enhanced-MRI (CE-MRI) to diagnose 13 cases of unilocular, round radiolucent lesions visualized on panoramic radiograph and/or CT. In the cases of UA, low signal intensity was observed on T1-weighted images (WIs), and a markedly high signal intensity was observed on T2-WIs; moreover, relatively thick rim-enhancement with/without small intraluminal nodules was observed upon CE-T1WIs. CE-MRI was considered useful in the diagnosis of UA, as characteristic features of this type of lesion i.e., thick enhancement of the tumor wall and small intraluminal nodules were detected only by CE-MRI in the present study.

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.

Effects of histone deacetylase inhibitor FR901228 on the expression level of telomerase reverse transcriptase in oral cancer.

We speculated whether or not the expression level of telomerase reverse transcriptase (hTERT) would be modulated by agents targeting epigenetics in oral cancer cell lines. Although hTERT is known to be targeted by epigenetic changes, it remains unclear how chemoagents targeting epigenetics work on hTERT transcription. In the present study, the epigenetic effects of the histone deacetylase (HDAC) inhibitor FR901228 on hTERT transcription in oral cancer cell lines were analyzed by RT-PCR. The mRNA expression of hTERT was upregulated after exposure to FR901228 in hTERT-negative Hep2 cells, and even SAS and KB cells expressed high levels of hTERT. Moreover, cotreatment of protein synthesis inhibitor cycloheximide (CHX) resulted in the induction of hTERT transcription by FR901228. This suggests that the induction of hTERT by FR901228 requires de novo protein synthesis to some extent and is more likely a direct than an indirect effect on epigenetic changes such as histone acetylation/deacetylation. We further examined the effect of FR901228 on c-myc protein, which is one of the main hTERT transcription activators. FR901228 repressed c-myc protein only in the absence of CHX, and depended on the enhancement of de novo protein synthesis. Our results indicate that c-myc protein is repressed indirectly by FR901228 but may not contribute to FR901228-induced hTERT transcription. The present study showed that the HDAC inhibitor FR901228 induced the hTERT gene by a complex mechanism that involved transcription factors other than c-myc, in addition to inhibition of histone deacetylation.

Role of O6-methylguanine-DNA methyltransferase and effect of O6-benzylguanine on the anti-tumor activity of cis-diaminedichloroplatinum(II) in oral cancer cell lines.

The DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) modulates the effectiveness of alkylating agents. However, the relationship between MGMT and the sensitivities to other agents has not been explored. In the present study, the association between MGMT expression and the cellular sensitivity to the platinum agent, CDDP was examined in four human oral cancer cell lines. CDDP depleted MGMT protein and mRNA levels in all four cell lines. Two cell lines with low MGMT expression were sensitive to an alkylating agent, N-methyl-N'-nitro-N-nitrosoguanidine and CDDP, whereas two other cell lines with high MGMT expression were resistant to both agents. Furthermore, the addition of the MGMT inhibitor, O6-benzylguanine (O6-BG), invariably enhanced CDDP sensitivity. CDDP depleted MGMT expression, and CDDP sensitivity was enhanced by O6-BG. These results provide valuable information about the relationship between MGMT expression and CDDP sensitivity in oral cancer chemotherapy.

Assessment of ameloblastomas using MRI and dynamic contrast-enhanced MRI.

We retrospectively evaluated magnetic resonance images (MRI) and dynamic contrast-enhanced MRI (DCE-MRI) of ameloblastomas. MRI and DCE-MRI were performed for 10 ameloblastomas. We obtained the following results from the MRI and DCE-MRI. (a) Ameloblastomas can be divided into solid and cystic portions on the basis of MR signal intensities. (b) Ameloblastomas show a predilection for intermediate signal intensity on T1WI, high signal intensity on T2WI, and well enhancement in the solid portion; they also show a homogeneous intermediate signal intensity on T1WI and homogeneous high signal intensity on T2WI, and no enhancement in the cystic portion. (c) The mural nodule or thick wall can be detected in ameloblastomas lesions. (d) CI curves of ameloblastomas show two patterns: the first pattern increases, reaches a plateau at 100-300 s, then sustains the plateau or decreases gradually to 600-900 s, while the other increases relatively rapidly, reaches a plateau at 90-120 s, then decreases relatively rapidly to 300 s, and decreases gradually thereafter. There was no difference in the CI curve patterns among primary and recurrent cases, a case with glandular odontogenic tumor in ameloblastoma or among histopathological types such as plexiform, follicular, mixed, desmoplastic, and unicystic type.

Assessment of carcinoma in the sublingual region based on magnetic resonance imaging.

In the present study, we attempted to diagnose and detect the extent of tumors in the sublingual region using magnetic resonance imaging (MRI) and dynamic MRI. MRI with or without gadopentetate dimeglumine (Gd-DTPA)-enhancement in seven lesions of the sublingual regions was performed. The seven lesions included four cases of mucoepidermoid carcinoma (mucoepidermoid Ca), two cases of adenoid cystic carcinoma (ACC), and one case of squamous cell carcinoma (SCC). Whether the tumor was malignant or benign, as well as the differential diagnosis, could not be determined on the basis of the MR signals, even when enhancement was performed. Dynamic MRI was performed in five cases, two cases of ACC, two cases of mucoepidermoid Ca, and one case of SCC. The dynamic MRI showed a rapid enhancement at 30-45 sec in all five cases before the normal sublingual gland began to be enhanced. The early phases at 30-45 sec of the dynamic MRI in five cases showed marked enhancement before the normal sublingual glands were enhanced, and therefore could clearly show the extent of the lesions. In conclusion, the dynamic MRI may be useful in differentiating malignant from benign tumors, and in detecting the extent of the tumors in the sublingual carcinomas.

Adenoviral p53 gene therapy in head and neck squamous cell carcinoma cell lines.

We reconstructed the recombinant p53-expressing adenovirus and examined its infections and effects in head and neck squamous cell carcinoma cell lines. Eight human head and neck squamous cell carcinoma cell lines were infected by the recombinant adenovirus harboring the lacZ gene (AxCAiLacZ) or the wild-type p53 gene (AxCAip53), and the effects were investigated. The eight cell lines were successfully infected by AxCAiLacZ at a level of more than 50%. The survival of all 8 squamous cell lines were inhibited in the range from 8 to 26.7% by only one treatment of the AxCAip53 infection. This result suggested that p53 gene therapy might become a useful tool in head and neck squamous cell carcinoma treatment.

Thermoradiotherapy combined with adenoviral p53 gene therapy in head and neck squamous cell carcinoma cell lines.

We examined effects of recombinant p53-expressing adenovirus combined with thermoradiotherapy in 8 head and neck squamous cell carcinoma (SCC) cell lines to improve the outcomes of the treatment of advanced head and neck cancer. The p53 gene therapy did not improve the discrepancy between thermoradiosensitivities among the 8 SCC cell lines. However, p53 gene therapy improved the effects of thermoradiotherapy in all 8 cell lines, and there were significant differences in four situations of the HSC4 44 degrees C (p=0.032), SAS at 44 degrees C (p=0.029), the KB at 43 degrees C (p=0.025), and the Ca9-22 43 degrees C (p=0.020). In comparing the survival rates of thermoradiotherapy with those of thermotherapy and radiotherapy, thermoradiotherapy demonstrated actual survival rates less than theoretical survival rates based on the survival rates of thermotherapy multiplied by the survival rates of radiotherapy in almost all treatments of thermoradio-gene therapy of the 8 SCC cell lines. These results demonstrate that thermoradiotherapy combined with p53 gene therapy may be a useful tool in treating SCC cells.

Effects of p53 gene therapy in radiotherapy or thermotherapy of human head and neck squamous cell carcinoma cell lines.

We report herein the effects of p53 gene therapy in the radiotherapy or thermotherapy of eight human head and neck squamous cell carcinoma (SCC) cell lines. The discrepancy between radiosensitivity combined with p53 gene therapy than that without p53 gene therapy increased among the eight SCC cell lines. The discrepancy increased in the thermosensitivity at 43 degrees C and decreased in that at 44 degrees C among the eight SCC cell lines. Thus, the p53 gene therapy did not always improve the discrepancy between radiosensitivity and thermosensitivity in the eight SCC cell lines. In the radiotherapy combined with adenoviral p53 gene therapy, the survival rates of three of eight SCC cell lines decreased, and that of only one cell line increased compared with radiotherapy alone. In thermotherapy combined with p53 gene therapy, the survival rates of three at 44 degrees C and five at 43 degrees C of the eight SCC cell lines decreased, although only one cell line at 43 degrees C increased its survival rate compared with thermotherapy alone. The p53 gene therapy decreased the survival rates of both radiotherapy and thermotherapy in three of eight SCC cell lines. Further, the distribution of plots on the basis of the time for 10% survival of radiotherapy and the dose for 10% survival of thermotherapy with p53 gene therapy shifted to the lower left side of the plots compared with those without p53 gene therapy. These findings indicated that p53 gene therapy improves the effects of both radiotherapy and thermotherapy.

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.

Thermochemotherapy of a human salivary adenocarcinoma cell line.

We report on thermochemotherapy in a human salivary gland adenocarcinoma cell line. Hyperthermia reduced the survival rate to 50 and 20% by heating at 43 degrees C for 40 and 60 min, respectively, and is by itself useful in human salivary gland carcinoma treatment. Adriamycin, cisplatin, and mitomycin C can possible be used clinically, while bleomycin and 5-fluorouracil cannot, to treat this carcinoma. The optimal temperature was considered to be 41 degrees C in adriamycin, 42 degrees C in cisplatin, 37 degrees C in mitomycin C, and 42 degrees C in bleomycin in the thermochemotherapy. Thermochemotherapy is a useful tool in the treatment of human salivary gland carcinoma cells, but it is necessary to select the best anticancer drugs and the optimal temperature for optimal success using this treatment.

Thermoradiotherapy combined with p53 gene therapy of human salivary gland adenocarcinoma cell line.

We report on thermoradiotherapy combined with p53 gene therapy in the human salivary gland adenocarcinoma cell line HSG. HSG cells were successfully infected at a rate of 62.3% with MOIs of 20 of either AxCAip53 or AxCAiLacZ. The AxCAiLacZ did not inhibit the survival of the HSG cells. The survival fractions of AxCAip53-infected HSG cells were lower than those of the AxCAiLacZ-infected HSG cells, but there was no significant difference (p=0.30). AxCAip53 decreased the survival rates of thermotherapy (43 degrees C; p=0.084 and 44 degrees C; p=0.18), radiotherapy (6 Gy; p=0.20) and thermoradiotherapy (6 Gy plus 43 degrees C; p=0.24 and 6 Gy plus 44 degrees C; p=0.96), but there were no significant differences. In comparing the survival rates of thermoradiotherapy with those of thermotherapy and radiotherapy, thermoradiotherapy, regardless of the combination with p53 gene therapy, demonstrated actual survival rates lower than theoretical survival rates based on survival rates of thermotherapy multiplied by survival rates of radiotherapy. This result indicates that thermoradiotherapy is effective in the treatment of HSG cells. Thermoradiotherapy combined with p53 gene therapy was the most effective therapy among the combinations of therapies demonstrating that thermoradiotherapy combined with p53 gene therapy may be a useful tool in the treatment of HSG cells.

Late course accelerated hyperfractionation radiotherapy concomitant with cisplatin in patients with esophageal carcinoma.

The purpose of this study was to examine whether combined cisplatin and late course accelerated fractionation radiotherapy (LCAH) (the combined group) can confer a better outcome over LCAH alone in treating esophageal carcinoma. Eighty-one eligible patients were randomly entered into two groups: the LCAH alone group, and the combined group. There was a better outcome in the combined group compared to the LCAH group, but the difference was not significant. The combined therapy could significantly improve the survival rate and local control of the disease in the early stage, but not in the advanced stage, because of the possibility of invasion. More severe complications occurred in the combined group than in the LCAH group, but they were within tolerance. In conclusion, LCAH concomitant with cisplatin could improve the survival of patients with esophageal cancer, especially in the early stage, as long as the side effects of the treatment are within tolerable limits.

Evaluation of tumor proliferation using dynamic contrast enhanced-MRI of oral cavity and oropharyngeal squamous cell carcinoma.

We investigated the relationship between the enhanced patterns acquired by dynamic MRI and the tumor cell proliferation estimated by immunostaining proliferating cell nuclear antigen (PCNA) in oral squamous cell carcinoma (SCC). Thirty patients with primary oral SCC underwent dynamic contrast enhanced (DCE)-MRI using a three-dimensional fast imaging with steady-state precession sequence. Tumor cell proliferation of all surgical specimens was evaluated using immunohistochemical staining with the anti-PCNA antibody. The relationship between the dynamic MRI parameters (maximum CI and maximum CI gain) and the PCNA labeling index was statistically analyzed using regression analysis. The time contrast index curves of all cases showed a rapid and high uptake pattern. The PCNA labeling index showed a significant correlation with maximum CI and maximum CI gain (P<0.0001, r=0.866 and P=0.0019, r=0.544, respectively). The assessment of DCE-MRI parameters may provide valuable information for tumor cell proliferation of the patients with oral cancer.

Assessment of pleomorphic adenomas using MRI and dynamic contrast enhanced MRI.

We evaluated magnetic resonance images (MRI) and the value of dynamic contrast enhanced MRI (DCE-MRI) of pleomorphic adenomas retrospectively. MRI was performed for 18 pleomorphic adenomas, including 11 cases with DCE-MRI. We obtained the following results on the MRI and DCE-MRI. (a). Pleomorphic adenomas showed a predilection for homogeneous intermediate signal intensity on T1-weighted images (T1WI), heterogeneous high signal intensity on T2-weighted images, and heterogeneous enhancement on Gd-T1WI. (b). Of 11 contrast index (CI) curves of pleomorphic adenomas, nine CI curves (81.8%) increased gradually to 600 s or increased gradually, reached a plateau, and sustained the plateau to 600 s. The remaining two (18.2%) increased gradually and decreased gradually thereafter. (c). CI curves reached the maximum CI index at 135-300 s.

Application of dynamic contrast-enhanced MRI to differentiate malignant lymphoma from squamous cell carcinoma in the head and neck.

Because malignant lymphoma, the second most common malignant tumor of the head and neck, and squamous cell carcinoma (SCC), the most common malignant tumor of the head and neck, require different treatments, it is important to be able to differentiate them. In the present study, we attempted to differentiate malignant lymphomas from SCCs using dynamic contrast-enhanced MRI (DCE-MRI). Seventeen lesions (in 8 cases) of malignant lymphoma and 30 cases of SCC were compared by DCE-MRI. Thirteen of 17 malignant lymphomas (76.5%) showed the maximum contrast index (CI) at 90-180 s, while 26 of 30 SCCs (86.7%) showed the maximum CI at a much faster 60-105 s. There was a statistically significant difference between SCC and malignant lymphoma in the time needed reach the maximum CI (p = 0.0177). There was also significant difference between SCC and malignant lymphoma in their maximum CIs (p < 0.001), with the maximum CIs of 29/30 SCCs (96.7%) above 2.0, while 12/17 malignant lymphomas (70.6%) showed CIs of less than 2.0. We consider these findings to be useful for distinguishing lymphomas from SCCs.

Assessment of dynamic MRI of Warthin's tumors arising as multiple lesions in the parotid glands.

We examined three patients with multiple synchronous Warthin's tumors in the bilateral parotid and described the value of using dynamic MRI. The time course of the contrast index (CI curves) was calculated from a dynamic series. Warthin's tumors showed intermediate signal intensity on T1WI, heterogeneous high and intermediate signal intensity on T2WI and a slight enhancement on Gd-T1WI. Warthin's tumors of CI curves showed specific findings. CI curves in each lesion showed the same pattern. It was difficult to diagnose masses as Warthin's tumors using only MR images on T1, T2 and Gd-T1WI. Dynamic MRI can distinguish Warthin's tumors from other possible tumors except for oncocytoma. Therefore, the use of dynamic MRI is recommended as a diagnostic method for Warthin's tumors in multiple synchronous lesions of the parotid gland.

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.

Thermoradiotherapy in human head and neck squamous cell carcinoma cell lines.

Thermoradiosensitivity of 8 cell lines of head and neck squamous cell carcinoma (HO-1-u-1, HSC2, HSC3, HSC4, SAS, KB, Hep2, and Ca9-22) was investigated. The differences of radiosensitivity between the cell line with the highest radiosensitivity and the cell line with the lowest radiosensitivity were 1.7-, 7.7-, and 41-fold at 2, 6 and 8 Gy, respectively. The differences between the cell line with the highest thermosensitivity and the cell line with the lowest thermosensitivity were 2.4-, 6.2- and 34.4-fold at 43 degrees C for 40, 60 and 100 min, and 2.6-, 4.9- and 127-fold at 44 degrees C for 20, 30 and 50 min, respectively. These findings indicated that there were large differences in both radiosensitivity and thermosensitivity among the 8 cell lines. There was a negative relationship between radiosensitivity and thermosensitivity (43 degrees C: r=-0.600, 44 degrees C: r=-0.848) in 7 of 8 cell lines, the exception being the HSC4 cell line, which was resistant to both therapies. Four of the 8 cell lines at 43 degrees C and 5 at 44 degrees C in the radiotherapy combined with thermotherapy showed actual survival rates smaller than the theoretical survival rates. Thus, thermoradiotherapy was deemed effective in the head and neck carcinoma cell lines, although 1 of 8 cell lines was resistant to both radiotherapy and thermotherapy.

Adenomatoid odontogenic tumor: correlation of MRI with histopathological findings.

Adenomatoid odontogenic tumor is a rare benign and odontogenic tumor that is frequently misdiagnosed as other odontogenic cysts and tumors on radiographic examination. To acquire additional information of adenomatoid odontogenic tumor, we performed magnetic resonance imagings (MRI) at a case of adenomatoid odontogenic tumor. The lesion was divided between the peripheral portion with a thick circular shape and the central portion with a round shape on the basis of the signal intensity (SI) of MRI. The peripheral portion showed intermediate SI contained multifocal no SI on T1WI, high SI contained multifocal no SI on T2WI, and heterogeneous enhancement on CE-T1WI. These multifocal areas corresponded to the numerous punctate radiopaque foci shown on computed tomography. The central portion showed homogeneous low SI on T1WI, homogeneous very high SI on T2WI, and no enhancement on CE-T1WI. Macroscopic examination revealed the round shaped lesion included one large cystic space correspondent to the central portion with a clear cystic wall correspondent to the peripheral portion on MRI. The MRI features corresponded to the macroscopic findings of the histopathological examination.