Expression of transforming growth factor-alpha in adenoid cystic carcinoma of the salivary gland.

BACKGROUND: Transforming growth factor-alpha (TGF-alpha) is expressed in both normal and malignant epithelial cells. Although the expression of TGF-alpha has been extensively studied in some human epithelial neoplasms, its expression in adenoid cystic carcinomas (ACCs) of the salivary gland has not been reported. METHODS: This study used an immunohistochemical technique to assess the expression of TGF-alpha in 40 ACC specimens from the salivary glands, seven specimens from labial minor salivary glands adjacent to mucoceles, and five specimens from normal submandibular glands. RESULTS: In the normal submandibular gland specimens, positive TGF-alpha staining was found in all ductal segments as well as in serous and some mucous acinar cells. In labial minor salivary glands adjacent to mucoceles, positive TGF-alpha staining was found in ductal, serous acinar, serous demilune, and a few mucous acinar cells. Positive TGF-alpha immunostaining was observed in all five salivary gland ACC specimens with a solid histologic pattern, and 31 of the 35 salivary gland ACC specimens with a tubulo-cribriform histologic pattern. Overall, 36 of 40 ACCs of the salivary gland expressed TGF-alpha. Positive TGF-alpha staining was found in the salivary gland ACC specimens with solid, tubular, duct-like, and pseudocystic structures. No significant correlation was found between TGF-alpha expression in ACCs of the salivary gland and age, sex, primary cancer location, TNM status, clinical stage, prognosis, histologic type, perivascular or perineural invasion, focal necrosis of the tumor, or cellular atypia. CONCLUSIONS: This study demonstrated that ductal and acinar cells of the normal submandibular gland and of labial minor salivary glands adjacent to mucoceles are the major sites of TGF-alpha synthesis and secretion. Furthermore, TGF-alpha is biosynthesized in salivary gland ACC tumor cells, forming solid, tubular, duct-like, and pseudocystic structures. Our data suggest that TGF-alpha may play an important biologic role as a mitogen in the growth of salivary gland ACC.

Expression of epidermal growth factor in salivary adenoid cystic carcinoma.

This study used an immunohistochemical technique to assess the expression of epidermal growth factor (EGF) in 40 specimens of salivary adenoid cystic carcinoma (ACC), 7 specimens of labial glands adjacent to mucocele, and 5 specimens of normal submandibular glands. In normal submandibular glands, immunohistochemically detectable EGF was demonstrated in all ductal segments, including intercalated, striated, and excretory duct cells. No EGF positive staining was found in acinar compartments. including serous and mucous acinar cells. In degenerated labial glands adjacent to mucocele, no EGF staining was detected in the remaining acinar and ductal cells. In salivary ACCs, positive EGF immunostaining was observed in one of the 5 (20%) ACCs with a solid pattern and in 13 of the 35 (37.1%) ACCs with a tubular-cribriform pattern. The overall EGF expression rate in 40 salivary ACCs was 35%. Positive EGF staining was predominantly found in tubular structures in the tubular ACCs and in duct-like structures in large cribriform patterns or in the stroma of the cribriform ACCs. There was no significant correlation between EGF expression in salivary ACCs and any of the clinicopathological parameters including patient age and sex, cancer location, TNM status, clinical stage, histologic type, perivascular or perineural invasion, focal necrosis of tumor, and cellular atypia. We conclude that the duct segments of the normal submandibular gland are the sites of EGF synthesis and secretion. In degenerated labial glands adjacent to mucocele, EGF synthesis is completely inhibited. Furthermore, EGF is mainly biosynthesized in cells forming tubular or duct-like structures in tubular or cribriform salivary ACCs, and EGF may play a biologic role, particularly as a mitogen in salivary ACC growth.

Expression of interleukin-2 receptor by activated peripheral blood lymphocytes upregulated by the plasma level of interleukin-2 in patients with recurrent aphthous ulcers.

This study used flow cytometry to determine the peripheral blood lymphocyte subsets and a sandwich enzyme immunoassay to measure the plasma levels of interleukin-2 (IL-2) and soluble IL-2 receptor (sIL-2R) in 34 patients in different stages of recurrent aphthous ulcers (RAU) and in 32 age/sex-matched normal control subjects. In the exacerbation stage of RAU, a significant increase in the percentages of CD3+ (p < 0.001), CD4+ (p < 0.001), CD4+ IL-2R+ (p < 0.001), CD8+ IL-2R+ (p < 0.01) and IL-2R+ cells (p < 0.001), in the CD4+/CD8+ (p < 0.01) and CD4+/CD3+ CD8+ ratios (p < 0.01), and in the plasma level of IL-2 (p < 0.001) was found in the patients as compared with the levels in the normal control subjects. However, in the post-exacerbation stage of RAU, there was a significant decrease in the percentage of CD4+ cells (p < 0.05) and in the CD4+/CD8+ (p < 0.01) and CD4+/CD3+ CD8+ ratios (p < 0.001), as well as a significant increase in CD8+ cells (p < 0.001) in the patients, as compared with the levels in the normal control subjects. Because the CD4+, CD4+ IL-2R+ and CD8+ IL-2R+ cell counts and the plasma level of IL-2 increased simultaneously in the patients in the exacerbation stage of RAU, we suggest that the markedly increased plasma level of IL-2 may have been secreted by the increased number of activated CD4+ cells, and that the expression of IL-2R by activated peripheral blood lymphocytes was upregulated by the plasma level of IL-2 in patients with RAU. In addition, the increase and then decrease of the CD4+/CD8+ ratio in the RAU patients and the increased number of CD4+ IL-2R+ and CD8+ IL-2R+ activated T cells in the RAU patients support the role of cell-mediated cytotoxicity in the immunopathogenesis of RAU.

Expression of proliferating cell nuclear antigen (PCNA) in oral submucous fibrosis, oral epithelial hyperkeratosis and oral epithelial dysplasia in Taiwan.

To test whether the oral epithelia of oral submucous fibrosis (OSF), epithelial hyperkeratosis (EH) and epithelial dysplasia (ED) may have increased proliferative activity under the long-term exposure to areca quid ingredients and whether there is an increased expression of proliferating cell nuclear antigen (PCNA) in oral premalignant lesions with disease progression, we used an immunohistochemical technique with the mouse monoclonal antibody PC10 to investigate PCNA expression in histologic sections of OSF, EH, ED and normal oral mucosa (NOM). Positive PCNA staining was found mainly in basal and parabasal epithelial cells in all specimens of OSF, EH, ED and NOM. The mean PCNA labeling indices (LI) in NOM, OSF, EH and ED were 8.8+/-2.7%, 22.1+/-12.5%, 25.5+/-5. 2% and 44.9+/-15.4%, respectively. Significant differences in the PCNA LI were noted between NOM and OSF (P<0.01), EH (P<0.001) or ED (P<0.001), as well as between ED and OSF (P<0.001) or EH (P<0.01). The gradual increase of PCNA expression with the morphologic transformation of normal epithelial cells into dysplastic epithelial cells suggests that there is increased proliferative activity in oral premalignant lesions with disease progression. However, no significant correlation was found between PCNA LI in OSF epithelium and the clinicohistologic parameters of OSF. In addition, the mean PCNA LI of p53-positive OSF cases (23.7+/-12.0%) was very close to that of p53-negative OSF cases (23.9+/-13.1%), suggesting that there was no association between PCNA and p53 expression in OSF.

Strong association of antiepithelial cell antibodies with HLA-DR3 or DR7 phenotype in patients with recurrent oral ulcers.

BACKGROUND: Previous studies showed that antiepithelial cell antibodies (anti-ECA) were present in 71% (15/21) of patients with recurrent oral ulcers (ROU) and that there was a strong association of human leukocyte antigen (HLA)-DRw9 with ROU in Chinese patients. In this study, we assessed anti-ECA in a larger group of Chinese patients with ROU (n = 88) in order to further investigate the association of anti-ECA with HLA-DR and -DQ antigens. METHODS: The anti-ECA in the sera of ROU patients were detected by an indirect immunofluorescence technique with rat esophagus as the substrate, and the HLA-DR and -DQ antigens in ROU patients were typed by a standard microcytotoxicity assay using Terasaki's oriental tray. RESULTS: The rate of anti-ECA positivity was significantly higher (p < 0.0001) in ROU patients (68%) than in healthy control subjects (0%). Furthermore, the rate of anti-ECA positivity in patients with major or minor oral ulcers (72%) was significantly higher (29%) than that in patients with herpetiform ulcers (p < 0.05). There was a significant increase in the frequency of DR3 or DR7 antigen expression (p < 0.0001, pc [p corrected] < 0.001, relative risk [RR] = 4.3, etiologic fraction = 0.41) in anti-ECA-positive ROU patients compared with the corresponding frequencies in healthy control subjects. There was also a significant increase in the frequency of DR7 or DRw9 antigen expression (p < 0.005, pc < 0.05, RR = 4.7, etiologic fraction = 0.45) compared to healthy controls. CONCLUSIONS: Because only DR3 or DR7 antigen occurred more frequently in anti-ECA-positive than in anti-ECA-negative ROU patients (p < 0.0007, pc < 0.05, RR = 19.6, etiologic fraction = 0.51), we concluded that the gene coding for DR3 or DR7 antigen may contribute to the presence of anti-ECA in Chinese patients with ROU.

Expression of p53 protein in oral submucous fibrosis, oral epithelial hyperkeratosis, and oral epithelial dysplasia.

BACKGROUND AND PURPOSE: In our previous study, positive p53 staining was observed in 47 of 81 (58%) cases of oral squamous cell carcinoma associated with areca quid (AQ) chewing and cigarette smoking. This study looked for expression of p53 protein in premalignant oral lesions in patients who chewed AQ or smoked cigarettes, or both. METHODS: Expression of p53 protein was examined in histologic sections of oral submucous fibrosis (OSF, n = 50), epithelial hyperkeratosis (EH, n = 10), epithelial dysplasia (ED, n = 10), and normal oral mucosa (NOM, n = 10) with antibodies against p53 protein using an immunoperoxidase technique. RESULTS: Positive p53 staining was observed in 30 (60%) OSF specimens, four (40%) EH specimens, seven (70%) ED specimens, and none of the NOM specimens. Only four (8%) of the OSF specimens and none of the EH specimens had more than 25% p53-positive keratinocytes. However, in four (40%) of the ED specimens, more than 50% of the keratinocytes were p53-positive. The degree of p53 staining increased with the morphologic transformation of normal-appearing epithelial cells into dysplastic epithelial cells. There was no significant correlation between expression of p53 in OSF epithelium and the clinicohistologic parameters of patients with OSF. CONCLUSIONS: These results demonstrate that p53 is often present in precancerous lesions of patients who chew AQ and smoke cigarettes. We suggest that p53 may play a role in dysplastic cell transformation in premalignant oral lesions.

Effects of continuous calcitonin treatment on osteoclasts derived from cocultures of mouse marrow stromal and spleen cells.

BACKGROUND: Continuous calcitonin (CT) treatment for bone diseases associated with increased bone resorption may be followed by prolonged depression of osteoclast response to CT. The mechanisms of this "escape" phenomenon remain unclear. METHODS: We examined the effects of continuous CT treatment on cell formation, calcitonin receptor (CTR) expression, response to CT, and bone resorption of osteoclasts in a coculture of mouse marrow stromal and spleen cells in the presence of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. Cells were cocultured and treated with salmon CT (sCT) for 7, 14, or 21 days. The effects of continuous CT treatment on osteoclast formation was determined by quantitation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs). CTR expression in osteoclasts was determined by binding of [125I]sCT in autoradiography. Bone resorption and CT responsiveness were assessed by examining the formation of resorption pits and by enumerating osteoclast reattachment on dentine slices after sCT rechallenge. RESULTS: TRAP-positive MNCs appeared in cocultures treated with sCT and were similar in number and morphology to those in control cultures, regardless of the concentration and duration of sCT treatment. A decrease in CTR expression was identified as a loss of silver grains from the TRAP-positive cells in cocultures receiving sCT treatment for 14 or 21 days. Partial recovery of CTR expression in TRAP-positive cells was evident in cocultures treated with sCT for only the first 7 days of coculture. TRAP-positive MNCs in cocultures treated with sCT for 14 or 21 days were resistant to the rechallenge with sCT. They attached to dentine slices and caused numerous resorption pits compared with control cells and cells treated with sCT for the first 7 days of coculture (p < 0.01). CONCLUSION: These findings suggest that the escape phenomenon that develops after continuous CT treatment may be due, at least in part to: 1) loss of responsiveness to CT in existing osteoclasts; and 2) development of new osteoclasts that are CTR-deficient and, therefore, refractory to CT rechallenge.