A clinicopathologic study of 196 intraoral minor salivary gland tumours

We present a retrospective study of 196 patients with intraoral minor salivary gland tumours, 128 malignant and 68 benign, diagnosed from 1954 to 1993 in the A.C. Camargo Hospital, São Paulo, Brazil. Sixty-five percent of the cases occurred in the palate, followed by tongue (9.7%) and retromolar area (6.1%). Pleomorphic adenoma was the most common benign tumour, and mucoepidermoid carcinoma was predominant among the malignant tumours. Surgery was the main treatment method and postoperative radiotherapy and radiotherapy alone were used in 40 and 15 patients, respectively. Local recurrence was observed in two patients with pleomorphic adenoma and in eight patients with malignant tumours. Regional lymph node metastases occurred in four cases and distant metastases in five. Forty-six of 47 patients with benigh tumours who were followed up from 1 to 7 years were alive without disease. Twenty-four of 79 patients with malignant tumours who were followed up for at least 5 years died due to the tumour and 47 were alive without disease.     

Salivary gland tumours treated in the stomatological clinics in Bratislava.

During the period of 1951-1996 a group of 1021 patients (484 men and 537 women, mean age 53 years, range 2-87 years) with salivary gland tumours were analysed retrospectively. The mean follow-up period was 8 years (range 10 months-25 years). The frequency of benign tumours was 74% (n = 755) and malignant tumours 26% (n = 266). Lesions were sited in the parotid gland 83% (n = 847), in the submandibular gland 10.8% (n = 110), in the sublingual gland 3.2% (n = 33) and in the minor salivary glands 3% (n = 31). The most common benign tumours were pleomorphic adenoma in 53.9% (n = 550) and Warthin's tumour 9.7% (n = 99). Of the malignancies, the adenoid cystic carcinoma was most common (6.4% of cases, n = 65) and mucoepidermoid carcinoma occurred in 5.2% (n = 53). The predominant therapy was surgery alone or in combination with postoperative radiotherapy in 93.7% (957 cases), radiotherapy alone after fine needle aspiration biopsy for 4.7%, and 19 patients remained untreated.     

Expression and mutations of p53 in salivary gland tumours

A series of 219 salivary gland tumours (103 carcinomas and 116 benign tumours) were analysed for p53 protein expression using immunohistochemistry, and for mutations in p53 gene using non-radioactive single strand conformation polymorphism (SSCP). p53 expression was present in 36% (42/116) of the benign tumours and in 54% (56/103) of the carcinomas. The highest prevalence of p53 expression was found in adenoid cystic carcinomas (69%). followed by mucoepidermoid carcinomas (67%). Of the benign tumours, pleomorphic adenomas showed the highest prevalence of p53 positivity (41%). In malignant tumours, expression of p53 bore no correlation to local recurrence, metastatic disease or survival of the patients. Exons 5 through 9 were analysed and four mutations were found in 20 cases of p53-immunopositive tumours and two in 20 p53-negative tumours. Each of the exons 5.6 and 8/9 had two mutations, whereas no mutations were detected in exon 7.     

Accessory parotid gland tumours: 24 years of clinical experience

The accessory parotid gland (APG) is salivary tissue anterior to and anatomically separate from the parotid gland. APG is a common anatomical variation, but APG tumours are extremely rare. The authors report 6 patients with APG tumours emphasizing the diagnosis, clinical features, indications and rationales for different treatment approaches. Patients with primary tumours of the parotid gland or APG tumours who underwent surgical treatment were included. APG tumours comprised 1.23% of overall parotid tumours (6/488) and had a malignancy rate of 33.3% (2/6). There were three male and three female patients with a mean age of 39 years (range 14–70 years). 5 of 6 parotidectomies entailed superficial lobectomy, while one was a total parotidectomy with composite resection of masseter muscle. Concomitant selective lymphadenectomy was carried out in 3 of 6 patients. At 5 years disease-free survival was 83.3%. Mean follow-up was 161 months (range 14–253 months). Although nonsalivary diagnoses frequently occur in the buccal area, APG tumours should be considered in every differential diagnosis in patients presenting with a mid-cheek mass. From oncosurgical, cosmetic and functional standpoints, treatment by facelift parotidectomy or ‘S-incision’ with concomitant superficial lobectomy is the recommended surgical approach; high-grade malignancies require total parotidectomy with regional lymphadenectomy.     

抑癌基因PTEN、p27及Ki-67在涎腺粘液表皮样癌中的表达

目的　检测抑癌基因PTEN、p2 7及Ki 6 7在涎腺粘液表皮样癌中的表达 ,并探讨其意义。方法　采用免疫组织化学SP法对 31例涎腺粘液表皮样癌的PTEN、p2 7和Ki 6 7表达进行检测 ,并作统计学分析。结果　PTEN、p2 7表达在高分化组粘液表皮样癌的阳性率分别为 88 89%、94 4 4 % ,低分化组分别为 5 3 85 %、4 6 15 % ;Ki 6 7增殖指数在高、低分化组间分别为 (5 18± 2 97) %、(10 38± 3 6 5 ) % ,三者在高低分化组间均有统计学差异 (P <0 0 5 ) ;PTEN和p2 7的表达呈正相关 (rs=0 4 4 5 ,P <0 0 5 ) ,p2 7和Ki 6 7的表达呈负相关 (rs=- 0 4 2 3,P <0 0 5 )。结论　涎腺粘液表皮样癌中 ,PTEN、p2 7和Ki 6 7的表达水平与其组织分化程度有关     

Predicting recurrence after oral precancer treatment: use of cell cycle analysis

Prediction of the behaviour of oral precancerous lesions (OPLs) is unreliable in clinical practice. The aim of this study was to analyse the efficacy of cell cyclin markers A and B1, and the proliferative marker Ki67, in predicting clinical outcome for patients with OPLs. A cohort of previously-treated patients with single OPLs were retrieved from the MaxilloFacial Dysplasia database and reviewed. All had dysplastic lesions excised by laser and were followed up for 5 years post-treatment. Outcome was determined as no recurrence or further disease. Excision specimens were re-examined immunohistochemically and labelling indices (LIs) for cyclin A, B1 and Ki67 determined. Forty patients, aged between 31 and 91 years, were recruited. There were no differences in age or sex. OPLs were predominantly leukoplakias on the floor of mouth or ventro-lateral tongue (65%), most of which exhibited moderate or severe dysplasia. Cyclin A LIs ranged from 3.9% to 31.3%, B1 0 to 28.3% and Ki67 3.5% to 54.5%. Using median LIs as 'cut off points' (12% cyclins; 22% Ki67) Kaplan-Meier survival analysis showed a significant risk of further progression of disease in patients with OPL LIs exceeding median values (Cyclin A p=0.02, Cyclin B1 p=0.01, Ki67 p=0.025). By combining analysis of both Cyclin A and B LI, the significance of the difference was increased (p<0.01). Cell cycle analysis is effective in identifying patients at risk of further progression of disease following treatment of OPLs. Multi-centre, longitudinal trials are needed to assess the precise role of cell cycle markers in their management.     

Flow cytometric S-phase fraction contributes to diagnosis of diploid malignant salivary gland tumours

DNA ploidy studies on salivary gland tumours have shown that the proportion of aneuploid cases, although confined to the malignant entities, is considerably lower than for other solid malignancies. To analyse whether the S-phase fraction (SPF) may contribute to discrimination of diploid malignant from benign tumours, DNA flow cytometric data from 45 different malignant salivary gland tumours was compared with that of 121 pleomorphic adenomas. All benign tumours were diploid. Twelve malignant tumours contained aneuploid cell populations. The SPF values for diploid malignancies ranged between 0.9% and 11.0% (mean 3.9%), and between 0.5% and 7.9% (mean 2.7%) for pleomorphic adenomas. A 4% cut-off value gained statistical significance for discriminating diploid malignant tumours from pleomorphic adenomas (P<0.01). The sensitivity for SPF>4% was 46% and the positive predictive value was 40%. A sensitivity of 60% and a positive predictive value of 54% was achieved by combining aneuploidy and SPF>4%. These results show that DNA flow cytometry may contribute to diagnostic assessment in salivary gland tumours.     

The relationship of the histologic grade at the deep invasive front and the expression of Ki-67 antigen and p53 protein in oral squamous cell carcinoma

BACKGROUND: Although many histopathologic characteristics of oral squamous cell carcinoma (O-SCC) have been identified as prognostic factors, accurate, and unequivocal factors have not been clearly identified. The purpose of this study was to evaluate a potential association between the histologic grade of malignancy at the deep invasive front and the expression of Ki-67 antigen and p53 protein in O-SCC. METHODS: The expression of Ki-67 antigen and p53 at the invasive tumor front area of O-SCC was examined by immunohistochemistry of archived tissue from 62 cases. The mean age of patients was 60.7 years (range: 37-89) and the male-female ratio was 1.6:1 (38 men, 24 women). There were 20, 17, 14, and 11 cases classified as stage I to stage IV, respectively. The correlation between the intensity of immunostaining for Ki-67 antigen and p53 and the histologic grade of malignancy at the deep invasive front (invasive front grade, IFG) was analyzed. The expression of Ki-67 antigen and p53 in normal oral epithelia (10 cases) was also investigated. RESULTS: The mean Ki-67 labeling index (LI) in the O-SCC samples was 32.8 +/- 12.0% (n = 62). The mean total score of IFG (IFG score) was 9.1 +/- 2.7 points (n = 62). There was a significant linear correlation between the IFG score and the Ki-67 antigen (gamma = 0.651, R2 = 0.596, P < 0.0001). Of 50 tumors examined, 27 (54.0%) exhibited p53-positive nuclear immunostaining. The staining patterns for Ki-67 antigen and p53 were similar. Both Ki-67-LI and p53-positive status were significantly correlated with the IFG scores. CONCLUSION: The findings of this study demonstrate that overexpression of Ki-67 antigen and p53 at the deep tumor invasive front of O-SCC is associated with histologic grade of malignancy.     

Proliferative,apoptotic and angiogenic potentials in jaws and long bones osteosarcomas: a comparative immunohistochemical study

J Oral Pathol Med (2010) 39 : 681–686 Background: Osteosarcomas (OS) of the jaws are uncommon lesions that represent less than 10% of all skeletal OS. It has a behavioral pattern which is less aggressive than their long bones counterparts. This study performed an immunohistochemical comparison between jaws and long bones OS. Methods: The study involved 15 jaws and 15 long bones OS tissue samples for the period from 1986 to 2005. Age, sex, histologic subtypes and grades were recognized. The samples were immunohistochemically stained with monoclonal antibodies to Ki‐67, P53 and vascular endothelial growth factor (VEGF). Results: The mean age of the patients with jaw OS was a decade higher than that of long bones OS. A slight male predominance in jaw OS was found (1.14:1), which was more pronounced in long bones OS (2:1). The chondroblastic subtype was the predominant in jaws (66.66%), whereas (60%) of long bones OS were of osteoblastic subtype. The Ki‐67 labeling index and the VEGF expression were significantly higher in long bones as compared with jaws OS, whereas there was no significant difference regarding the P53 expression between jaws and long bones OS. Conclusions: Jaws and long bones OS bear a comparable cell cycle dysregulation when quantified with P53 immunostaining, whereas the long bones OS have a higher proliferative and angiogenic capacity than their jaw counterparts when evaluated with Ki‐67 and VEGF immunoexpressions respectively.     

Ki-67 expression predicts radiosensitivity in oral squamous cell carcinoma

The prognostic relevance of Ki-67 expression in oral squamous cell carcinoma (OSCC) is still controversial. As proliferating cells are more susceptible to ionizing radiation, the authors investigated if a high proliferation rate reflected by Ki-67 expression, predicts radiosensitivity in OSCC patients. In 52 patients with OSCC who received primary surgery followed by radiation therapy, the proliferation rate was assessed by Ki-67 immunhistochemistry and correlated to recurrent free survival and overall survival. Low proliferative carcinomas showed a significantly shorter mean time to recurrence of 27.5 months compared to 49.5 months of high proliferative tumours (p=0.048). The 5-year survival rate of low proliferative tumours was 49% compared to 80% for high proliferative tumours (p=0.042). This study indicates that tumours with high proliferative activity are more susceptible to radiation therapy. Ki-67 might be used as a marker to predict the response to radiation therapy in patients with OSCC.     

The biological behavior of adenocystic carcinomas of the mouth cavity]

During the years 1965-1976, 19 patients with adenocystocarcinoma have been treated by the author on an inpatient basis. There were 15 cases with primary tumours and 4 cases of recurrences (2 of them with distant metastases). Up to now, 8 patients have died of the tumour. The mean length of survival of these patients was 51 months; that of the patients still alive, 84 months. These results lead to the conclusion that adenocystocarcinomata develop very slowly, affect mainly the accessory salivary glands and show lymphogenous and haematogenous metastasization even without previous recurrence. Radical surgical therapy of the primary tumour is advocated also in patients presenting with distant metastases.     

p^53,C—erbB—2癌基因蛋白在涎腺粘液表皮样癌中的表达及其临床 …

目的 探讨癌基因蛋白ｐ５３、Ｃ－ｅｒｂＢ－２（ｐ１８５）过度表达与粘液表皮样癌生物学行为的关系。方法 利用微波免疫组织化学方法对３２例人粘液表皮样癌的癌基因蛋白ｐ５３、Ｃ－ｅｒｂＢ－２进行检测。结果 正常涎腺组织ｐ５３、Ｃ－ｅｒｂＢ－２均为阴性反应。癌旁导管上皮细胞两者阳性率分别为１０．０％及１５．０％,但腺泡细胞阴性。粘液表皮样癌组织中ｐ５３、ＣｅｒｂＢ－２阳性率分别为４０．６％及４６．９％。ｐ     

Diagnosis and treatment of epithelial salivary gland tumours in children and adolescents

In a series of 2,871 epithelial salivary gland neoplasms managed in the Peking University School of Stomatology between 1974 and 1999, 86 arose in children <16 years of age (52 parotid, 12 submandibular gland, 2 sublingual gland, and 20 minor salivary gland). Considerable delay was encountered in diagnosis (benign 24 months and malignant 16 months). In this group of children, 46 tumours (53%) proved to be malignant, with an incidence in the parotid, submandibular, sublingual, and minor salivary glands of 31/52 (60%), 2/12, 0/2, and 13/20 (65%), respectively. Sixty-six of 86 neoplasms (77%) occurred in children between 10 and 16 years of age. Only six neoplasms were encountered in children of 5 years or younger, four of which were high-grade malignant tumours. Benign tumours were successfully treated by local excision with only one recurrence. Of 46 malignant neoplasms, 8 were treated palliatively; of the remainder 8 were lost to follow-up and 2 patients died of their disease.     

p53、C-erbB-2癌基因蛋白在涎腺粘液表皮样癌中的表达及其临床病理学意义

目的探讨癌基因蛋白p53、C-erbB-2（p185）过度表达与粘液表皮样癌生物学行为的关系。方法利用微波免疫组织化学方法对32例人粘液表皮样癌的癌基因蛋白p53、C-erbB-2进行检测。结果正常诞腺组织p53、C-erbB-2均为阴性反应。癌旁导管上皮细胞两者阳性率分别为10.0％及15.0％,但腺泡细胞阴性。粘波表皮样癌组织中p53、C-erbB-2阳性率分别为40.6％及46.9％。p53、C-erbB-2在粘液细胞、表皮样细胞及中间细胞内均有表达。p53、C-erbB-2的表达与粘液表皮样癌肿瘤组织学类型、分化程度及肿瘤复发等肿瘤生物学行为密切相关（P＜0.01）。结论p53、C-erbB-2可作为粘液表皮样癌的分化性标志物,用于监测病情、判断预后等。     

Intraoral salivary duct carcinoma: case report with immunohistochemical observations

Salivary duct carcinoma is an uncommon malignant salivary gland tumor that occurs predominantly in the parotid gland. Oral involvement is extremely rare, with few cases having been reported in the literature. The tumor is characterized by an aggressive behavior and has a poor prognosis. We describe a case of salivary duct carcinoma arising in the hard palate of a 63-year-old man. Immunohistochemical analysis revealed that tumor cells tested positive for cytokeratin, epithelial membrane antigen, proliferating cell nuclear antigen, Ki67, p53, laminin, and collagen IV. Despite radical surgical resection, bilateral neck dissection, and postoperative radiotherapy, liver metastases developed, and the patient subsequently died of his disease.     

PCNA, Ki-67 and p53 expressions in submandibular salivary gland tumours

Salivary gland tumours are uncommon with a broad heterogeneity. The most common benign tumour is the pleomorphic adenoma, whereas mucoepidermoid carcinoma and adenoid cystic carcinoma predominate among the malignancies. Most salivary gland tumours occur in the parotid, and consequently clinical and biological data are normally derived from this site. This work describes the expressions of PCNA, Ki-67 and p53 in 15 pleomorphic adenomas, 15 mucoepidermoid carcinomas and 15 adenoid cystic carcinomas of the submandibular gland. Our results showed that all pleomorphic adenomas were negative for p53 and Ki-67 with 66.6% being positive for PCNA. Conversely, p53 was positive in 53% of the mucoepidermoid carcinomas and in 20% of the adenoid cystic carcinomas. Ki-67 was expressed in 47.7% of the mucoepidermoid carcinomas and 40% of the adenoid cystic carcinomas. All malignant tumours were positive for PCNA. These results indicate that the proliferative rate analysed with PCNA and Ki-67 and the expression of p53 in pleomorphic adenoma and adenoid cystic carcinoma of the submandibular gland were similar to those described in the parotid and minor salivary glands. However, mucoepidermoid carcinomas showed higher expression of these markers than those of other salivary glands. This work is the first describing the expression of these immunohistochemical markers exclusively in submandibular salivary gland tumours.     

Nuclear DNA content, an adjunct to p53 and Ki-67 as a marker of resistance to radiation therapy in oral cavity and pharyngeal squamous cell carcinoma

Factors of prognosis and radioresistance in oral cavity and pharyngeal squamous cell carcinoma (OCPSCC) are limited. In the present study, the usefulness of tumor DNA content in predicting radioresistance in patients with OCPSCC has been investigated. Radioresistance has been defined as local recurrence or tumor persistence after radiation therapy. DNA-ploidy analysis was performed by static cytometry on smears of cell suspensions obtained from formalin-fixed paraffin-embedded material and stained with Feulgen. DNA-ploidy was correlated with the proliferation rate (Ki-67) and p53 protein accumulation obtained by immunohistochemistry. The follow-up of patients ranged from 8 to 62 months. Radioresistance was more common in non-diploid tumors; 14/28 (50%) non-diploid tumors recurred, whereas only 3 (10.7%) out of 28 diploid tumors had local failure (P=0.0019). Proliferation rate and p53 accumulation, evaluated by immunohistochemistry, also added prognostic information. Twelve out of 14 failures were from non-diploid tumors with a low proliferation rate (Ki-67<20%), whereas none of 20 p53-negative diploid tumors developed recurrences. This study showed that non-diploid tumors responded poorly to radiotherapy. DNA content appeared, therefore, as a significant prognostic marker for the evaluation of OCPSCC in patients receiving radiation therapy. This study also showed that DNA content adds information to p53 accumulation and the proliferation rate (Ki-67) for the purposes of determining patient management.     

Pulmonary metastases of malignant tumors of the oral and maxillofacial region. Analysis of 70 cases

Records of 685 cases of malignant tumors in the oral and maxillofacial regions were collected. Pulmonary metastases occurred in 70 of these cases. The metastatic rate was 10.2%. Roentgenographic appearances of pulmonary metastases were divided into 3 types: (1) Multiple ball-like lesion (78%); (2) Single ball-like lesion (14%); (3) Irregular lesion (8%). When metastasis was single, diagnosis was made according to the location of the metastasis and comparison between and old recent roentgenograms. Pulmonary metastasis was more frequent in tumors of the submandibular gland (34.5%), the base of the tongue (33.3%), and the sublingual gland (27.3%). Adenoid cystic carcinoma (47.8%) and adenocarcinoma (26.5%) might readily develop pulmonary metastasis. Pulmonary metastasis was rare in squamous cell carcinoma (2.5%). The metastatic rates were 3.9%, 5.4%, 6.5%, and 7.1% in stage I, II, III and IV, respectively. The period between the occurrence of pulmonary metastasis and initial treatment was from 1 month to 8 years and 5 months, with a mean of 33.8 months. The period between the occurrence of pulmonary metastasis and death of the patient was from 1 month to 8 years and 8 months, with a mean of 21.4 months.     

Connexin 43, Bcl-2, Bax,Ki67, and E-cadherin patterns in oral squamous cell carcinoma and its relationship with GJA1 rs12197797 C/G

Background To our knowledge, there is no useful and accurate prognostic biomarker or biomarkers for patients with oral squamous cell carcinoma (OSCC), a tumor with uncertain biological behavior, and unpredicTable clinical progress. The purposes of this study were: a) to determine the expresión profile of Connexin 43, Bcl-2, Bax, E-cadherin, and Ki67 in patients with OSCC; b) identify the GJCA1 rs12197797 genotypic composition.Material and Methods A cross-sectional study using genomic DNA and biopsy samples extracted from the oral mucosa with/without OSCC, older than 18 years, both genders, attended at Facultad de Odontología, Universidad Nacional Córdoba. Immunostaining for Cx43, Bcl-2, Bax, E-cadherin, and Ki67 and genotyping GJA1 rs12197797 by RFLP were performed. Odds Ratio (95% CI), Spearman Coefficient were estimated. Mann-Whitney test was applied to analyze immunostaining between controls/cases (p <0.05 was set for statistical significance).Results GG (mutant) was the most frequent genotype in patients with OSCC diagnosis (53.2%) in relation to CC “healthy” genotype (p=0.00487; OR=7.33; CI95% [1.1-54.7]). And, the allele G (mutant) had a presence in 75.5% of OSCC patients. However, no significant association was observed between alleles C/G and diagnosis (p=0.0565). The heterozygous genotype was the most frequent in the patients of both groups Cx43 and E-cadherin markers were lower in OSCCs in relation to controls. Ki67 and Bcl-2 immunolabeling were high on OSCC, and Bax immunomarker was diminished in OSCC.Conclusions We hypothesized that the oral epithelium losses Connexin 43 and E-cadherin in the membrane, which modifies cell differentiation. The Ki67 and Bcl2 overexpression would increase the cell density in the tissue, by promoting proliferation and decreasing apoptosis. And, this study shows evidence that patients who carry on allele G of GJA1rs12197797 could be at risk of developing OSCC. Key words:Cx43, E-cadherin, Ki67, Bax, Bcl-2, immunostaining expression profile, GJA1 rs12197797 genotyping.     

EGFR and Ki‐67 expression in oral squamous cell carcinoma using tissue microarray technology

J Oral Pathol Med (2010) 39 : 571–578 Objective: Our aim was to validate the use of tissue microarrays (TMA) in oral squamous cell carcinomas (OSCC) to analyse epidermal growth factor receptor (EGFR) and Ki‐67 expression. We also analysed the relationship that the expression of these markers may have with clinical, pathological and survival variables. Patients and methods: The study sample comprised 39 unselected patients diagnosed and treated for OSCC. We analysed Ki‐67 and EGFR expression by immunohistochemistry on formalin‐fixed, paraffin‐embedded surgical specimens. Whole sections (WS) were compared with double 1.5 mm core‐tissue microarrays. Results: High EGFR expression was observed both on TMA (in 98% of the cases) and WS (in 100% of the cases) with substantial agreement kappa value (0.720). EGFR expression was not significantly associated with clinical, pathological and survival variables on TMA and WS. Ki‐67 analysis showed a Spearman correlation of 0.741 with a Ki‐67 mean labelling index of 45% in TMA and 56.8% in WS. We found a significant relationship between gender and Ki‐67 labelling index on WS (P = 0.022) and TMA (P = 0.002). Clinical stage was the only parameter in multivariate analysis that had a significant predictive value. Conclusion: We demonstrate that dual 1.5 mm core TMA is a valid, rapid, economical and tissue‐saving way to study OSCC biopsies and that it presents strong correlation with the WS. EGFR overexpression in OSCC suggests that these tumours may be a candidate for therapy investigation directed to EGFR.