Transformation Of Oral Epithelial Dysplasia Research Articles

Similar Effect of P16 Hydroxymethylation and True Methylation on Prediction of Malignant Transformation of Oral Epithelial Dysplasia: A Prospective Study

Background: Total P16 methylation (P16M), including P16 hydroxymethylation (P16H) and true-P16M, correlates with malignant transformation of oral epithelial dysplasia (OED). Both true-P16M and P16H are early events in carcinogenesis. Aim: The aim of this study is to prospectively determine if discrimination of true-P16M from P16H similarly is necessary for prediction of cancer development from OEDs. Methods: Patients (n = 265) with mild or moderate OED were recruited into the double-blind 2-center cohort. Total-P16M and P16H were analyzed using the 115-bp MethyLight, TET-assistant bisulfite (TAB) methylation-specific PCR (MSP), and TAB-sequencing. Total-P16M-positive and P16H-negative samples were defined as true-P16M-positive. Progression of OEDs was monitored for a minimum 24 months follow-up period. Results: P16H was detected in 23 of 73 (31.5%) total-P16M-positive OEDs. Follow-up information was obtained from 247 patients with an ultimate compliance of 93.2%. OED-derived squamous cell carcinomas were observed in 13.0% (32/247) patients during the follow-up (median, 41.0 months). The cancer progression rate for total-P16M-positive patients was significantly increased when compared with total-P16M-negative patients (23.3% vs 8.6%; adjusted odds ratio = 2.67 [95% CI: 1.19-5.99]). However, the cancer progression rate was similar between P16H- and true-P16M-positive OEDs (26.1% [6/23] vs 22.0% [11/50]; odds ratio = 0.80 [95% CI: 0.22-2.92]). The progression-free survival was also similar for these patients. Conclusion: P16H and true-P16M are similar biomarkers for determining malignant potential of OEDs. Discrimination of P16H from true-P16M, at least in OED, may be not necessary in clinical applications.

A similar effect of P16 hydroxymethylation and true-methylation on the prediction of malignant transformation of oral epithelial dysplasia: observation from a prospective study

BackgroundTotal P16 methylation (P16M), including P16 hydroxymethylation (P16H) and true-P16M, correlates with malignant transformation of oral epithelial dysplasia (OED). Both true-P16M and P16H are early events in carcinogenesis. The aim of this study is to prospectively determine if discrimination of true-P16M from P16H is necessary for prediction of cancer development from OEDs.MethodsPatients (n = 265) with mild or moderate OED were recruited into the double blind two-center cohort. Total-P16M and P16H were analyzed using the 115-bp MethyLight, TET-assisted bisulfite (TAB) methylation-specific PCR (MSP), and TAB-sequencing. Total-P16M-positive and P16H-negative samples were defined as true-P16M-positive. Progression of OEDs was monitored for a minimum 24 months follow-up period.ResultsP16H was detected in 23 of 73 (31.5%) total-P16M-positive OEDs. Follow-up information was obtained from 247 patients with an ultimate compliance rate of 93.2%. OED-derived squamous cell carcinomas were observed in 13.0% (32/247) patients during follow-up (median, 41.0 months). The cancer progression rate for total-P16M-positive patients was significantly increased when compared to total-P16M-negative patients [23.3% vs 8.6%; adjusted odds ratio = 2.67 (95% CI: 1.19–5.99)]. However, the cancer progression rates were similar between P16H- and true-P16M-positive OEDs [26.1% (6/23) vs 22.0% (11/50); odds ratio = 0.80 (95% CI: 0.22–2.92)]. The cancer-free survival was also similar for these patients.ConclusionP16H and true-P16M are similar biomarkers for determining malignant potential of OEDs. Discrimination of P16H from true-P16M, at least in OED, may be not necessary in clinical applications.Trial registrationThis study is registered prospectively in the U.S. National Institutes of Health Clinical Trials Protocol Registration System (trial number NCT02967120, available at https://ClinicalTrials.gov/ct2/show/NCT02967120).

Coalition of E-cadherin and vascular endothelial growth factor expression in predicting malignant transformation in common oral potentially malignant disorders.

Background:Reduced E-cadherin expression and increased VEGF expression is known to be involved in tissue growth and transformation of Oral Potentially Malignant Disorders (OPMDs) and has been correlated with their differing histological grades in numerous studies.Aim:To evaluate Immunohistochemical (IHC) expression of both E-cadherin and VEGF in predicting the malignant transformation potential of common OPMDs.Materials and Methods:Ten cases each of Normal Oral mucosa (NOM), Mild Oral Epithelial Dysplasia (OED), Moderate OED, Severe OED, Oral Submucous Fibrosis, (OSMF) and Oral Squamous Cell Carcinoma (OSCC) were stained and evaluated for the expression of Ecadherin and VEGF. Quick score (QS) for expression intensity in all epithelial layers was calculated for both markers and results statistically analysed using Kruskal –Wallis ANOVA and Mann-Whitney “U” test.Results:E-cadherin expression was continuous and membranous in all the layers of NOM and reduced with progressing grades of OED to OSCC. In OSMF, expression was intermediate between moderate and severe OED. VEGF expression increased as the disease progressed from normal to increasing grades of OED to malignancy. In OSMF, expression was similar to that in mild OED. VEGF, E-cadherin expression for basal and parabasilar cells showed a strong statistically significant negative correlation in NOM. A very strong statistically significant positive correlation with perfect monotonic relation was noted in superficial cells in severe OED group and OSCC group.Conclusion:E-Cadherin and VEGF could be used as combination markers to predict the potential risk for malignant transformation in OEDs.

PITX1 is a reliable biomarker for predicting prognosis in patients with oral epithelial dysplasia

Paired-like homeodomain 1 (PITX1) genes are essential in human development. In the present study, PITX1 protein expression was evaluated in human normal oral mucosa, oral epithelial dysplasia and oral squamous cell carcinoma (OSCC), with the aim of examining the expression patterns of these critical genes during the multi-stage transformation of oral epithelial dysplasia to OSCC. PITX1 and Ki-67 expression were assessed by immunohistochemistry in 26 individuals with normal oral mucosa, 106 patients with oral epithelial dysplasia and 97 OSCC patients. The labeling indices (LIs) of PITX1 and Ki-67 were calculated and their correlation with the incidence of malignancy was evaluated. The PITX1 LI of the dysplasia specimens was significantly lower than that of the normal oral mucosa samples, but significantly higher than that of the OSCC samples. The oral epithelial dysplasia patients that exhibited low PITX1 expression showed a significantly higher incidence of malignant transformation than those exhibiting high PITX1 expression, regardless of the histological grades of their oral epithelial dysplasias. On the other hand, no correlation was observed between the Ki-67 LI and the incidence of malignancy. These results suggested that PITX1 suppression is associated with malignant transformation in the oral epithelium and that PITX1 expression may serve as a novel biomarker for predicting prognosis in oral epithelial dysplasia.

Malignant transformation of oral epithelial dysplasia: a real-world evaluation of histopathologic grading

This study describes the predictive value of oral epithelial dysplasia (OED) grading as an indicator for malignant transformation and progression. The records of an Australian-based pathology laboratory were searched for oral mucosal biopsies with a dysplastic or malignant diagnosis. Examination for an association with progression and malignant transformation without reinterpretation was performed. Analysis was undertaken using hazard ratios and the Fisher exact test. A total of 368 patients with a diagnosis of OED were included. Twenty-six patients (7.1%) underwent progression or malignant transformation; the annual malignant transformation rate was 1%. No other characteristics were associated with a heightened risk of progression or transformation. The severity of OED was not associated with risk of malignant transformation, suggesting that the current OED grading system is not useful for predicting patient outcomes or for determining management strategies. Definitive treatment of all OED is recommended, until a more reliable progression/transformation system is developed.

Evaluation of Microvascular Density as a Predictive Indicator for Malignant Transformation of Oral Epithelial Dysplasia (OED)

While not all OED lesions will transform, approximately one third of OED lesions progress to oral squamous cell carcinoma (OSCC). Provided the poor overall prognosis for OSCC patients, intervention at the premalignant stage of OED is an attractive clinical strategy. We do not, however, currently have biomarkers to identify the most aggressive premalignant lesions. Identification of suchbiological indicators would allow for targeting of OED lesions with high transformation risk and direct lesional-specific management e.g.

Overexpression of Smad proteins, especially Smad7, in oral epithelial dysplasias

Transforming growth factor β, via membrane-bound receptors and downstream Smad2-4, 7, can modulate tumorigenesis. Smad2 and Smad3 heterodimerize with Smad4, and the complex migrates to the nucleus to regulate the expression of target genes. Smad7 is a key negative regulator of this signaling pathway. This study aimed to examine Smad2-4, 7 expression and phosphorylated Smad2-3 (p-Smad2-3) in oral epithelial dysplasia and compared it with normal oral mucosa, hyperkeratosis/epithelial hyperplasia and squamous cell carcinoma (SCC). Immunohistochemical staining of Smad2-4, 7 and p-Smad2-3, was performed for 75 samples of human oral mucosa, including hyperkeratosis/epithelial hyperplasia (n = 20), mild epithelial dysplasia (n = 11), moderate to severe epithelial dysplasia (n = 11), and SCC (n = 43). Normal buccal mucosa samples (n = 9) were also included. A significant increase in Smad7 expression was observed in the ascending order of samples of normal oral mucosa, hyperkeratosis/epithelial hyperplasia/mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, and well-differentiated oral SCC/moderately to poorly differentiated oral SCC. Additionally, significant increases in Smad7 expression were noted as compared with expression of Smad2-4 and p-Smad2-3 in lesions of hyperkeratosis/epithelial hyperplasia, mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, well-differentiated oral SCC, and moderately to poorly differentiated oral SCC. Our results indicate that Smad proteins, particularly Smad7, in oral epithelial dysplasia and SCC could contribute to the attenuation of Smads anti-proliferative signaling in cancer development. Smad7 could be a marker for risk of malignant transformation of oral epithelial dysplasia.

The clinical determinants of malignant transformation in oral epithelial dysplasia

While the size and clinical appearance are known risk factors for malignant transformation of potentially malignant oral the importance of site, grade of dysplasia and exposure to environmental carcinogens remains controversial. We aim to report the clinical determinants of malignant progression in a series of patients with histopathologically graded oral epithelial dysplasia (OED). We recruited patients with a histopathological diagnosis of OED to a longitudinal observational study in a tertiary oral dysplasia clinic. Clinical, histopathological and risk factor data were recorded at baseline. One of three clinical endpoints were determined: malignant transformation, progression of dysplasia grade, remission/stable dysplasia grade. Ninety-one patients meeting the criteria gave consent for inclusion to the cohort, with outcomes reported after a median follow up of 48 months. An estimated 22% (SE 6%) of patients underwent malignant transformation within 5 years, with significant predictors being: non-smoking status (χ(2)=15.1, p=0.001), site (χ(2)=15.3, p=0.002), non-homogeneous appearance (χ(2)=8.2, p=0.004), size of lesion >200 mm(2) (χ(2)=4.7, p=0.03) and, of borderline significance, high grade (χ(2)=5.8, p=0.06). Gender, age, number of lesions and alcohol history did not predict for malignant transformation. Although a number of these clinical determinants have previously been associated with higher malignant transformation in OED, the high-risk nature of lesions in non-smokers is of particular note and requires a greater emphasis and recognition amongst clinicians dealing with OED. It suggests that those non-smokers with OED, have an inherited or acquired predisposition and should be treated more aggressively; these should form the focus for further investigation.

Malignant transformation of oral epithelial dysplasia: clinicopathological risk factors and outcome analysis in a retrospective cohort of 138 cases

To explore the usefulness of a new binary system of grading dysplasia proposed by the World Health Organization and to identify significant risk factors for malignant transformation in a long-term follow-up cohort of patients with oral epithelial dysplasia. A total of 138 patients with histologically confirmed oral dysplasia between 1978 and 2008 were reviewed retrospectively in our department. The mean follow-up period was 5.1 years. Of these dysplasias, 37 (26.8%) developed into cancer, with a mean duration of 4.6 years. Cox regression analysis revealed that high-grade dysplasia was an independent risk factor for transition, but age, gender, lesion site, diet habit, smoking and alcohol intake were not risk factors. High-grade dysplasia was associated with a 2.78-fold (95% confidence interval 1.44-5.38; P = 0.002) increased risk of transition, as compared with low-grade dysplasia. Consistently, high-grade dysplasia had a significantly higher incidence of malignancy than low-grade dysplasia by Kaplan-Meier analysis (log-rank test, P = 0.001). The utilization of high-grade dysplasia as a significant indicator for evaluating malignant transformation risk in patients with potentially malignant lesions is suggested; this may be helpful to guide treatment selection in clinical practice.

Increased ΔNp63 expression is predictive of malignant transformation in oral epithelial dysplasia and poor prognosis in oral squamous cell carcinoma

This study examined immunohistochemical expression of ΔNp63, a keratinocyte stem cell marker, in oral leukoplakia (OL) and oral squamous cell carcinoma (OSCC) and then to elucidate usefulness of ΔNp63 as a marker for diagnosis and prognosis. One-hundred and twelve cases of OL and 81cases of OSCC were analyzed by immunohistochemical staining for ΔNp63, Ki-67, and cytokeratin 14. These labeling indices (LIs) were calculated, and the association of these LIs with clinicopathologic characteristics in the OL and OSCC was evaluated. In the OL, these LIs increased significantly according to the severity of epithelial dysplasia (p<0.0001). ΔNp63-LI in the OL with malignant transformation was significantly higher than that in the OL without (49.3 vs. 34.2%; p<0.01). In the OSCC, the LIs increased significantly in association with the histologic grade (p<0.0001). A significant difference between the high and low ΔNp63-LI groups was found in the incidence of cervical lymph node and distant metastasis (p<0.05). The prognosis of the high ΔNp63-LI (mean value >73.8%) group is poorer than that of the low ΔNp63-LI (mean value ≤73.8%) group (p<0.05). These results suggested that increased ΔNp63 expression is involved in malignant transformation in epithelial dysplasia and poor prognosis in OSCC.

Methylation of p16 CpG Island Associated with Malignant Progression of Oral Epithelial Dysplasia: A Prospective Cohort Study

Inactivation of p16 gene by CpG methylation is a frequent event in oral epithelial dysplasia. To investigate the predictive value of p16 methylation on malignant potential in oral epithelial dysplasia, we carried out the prospective cohort study. One hundred one patients with histologically confirmed mild or moderate oral epithelial dysplasia were included in the present cohort study. p16 Methylation status of the oral epithelial dysplasia lesions from 93 cases was obtained by methylation-specific PCR. Progression of the oral epithelial dysplasia lesions was examined in 78 cases histologically during a 45.8 months follow-up period. The association between p16 methylation and progression of oral epithelial dysplasia was analyzed. Of the 93 enrolled cases, 15 cases were lost during the follow-up because of changes of contact information, with a compliance of 83.9%. p16 Methylation was detectable in oral epithelial dysplasia lesions from 32 (41.0%) of 78 enrolled patients. Oral epithelial dysplasia-related squamous cell carcinomas were observed in 22 patients (28.2%) during the follow-up. Rate of progression to oral cancer in patients with the p16-methylated oral epithelial dysplasia was significantly higher than that with the p16-unmethylated oral epithelial dysplasia (43.8% versus 17.4%; adjusted odds ratio, 3.7; P = 0.013), especially for patients at the baseline age of > or = 60 years (adjusted odds ratio, 12.0; P = 0.003) and patients with moderate oral epithelial dysplasia (adjusted odds ratio, 15.6; P = 0.022). The overall sensitivity and specificity of prediction of malignant transformation of oral epithelial dysplasia by p16 methylation were 63.6% and 67.9%, respectively. p16 Methylation was correlated with malignant transformation of oral epithelial dysplasia and is a potential biomarker for prediction of prognosis of mild or moderate oral epithelial dysplasia.

P16 promoter methylation predicts malignant transformation in oral epithelial dysplasia

p16 promoter methylation predicts malignant transformation in oral epithelial dysplasia

Evaluation of a new binary system of grading oral epithelial dysplasia for prediction of malignant transformation

The aim of this paper is to assess the reproducibility of a novel binary grading system (high/low risk) of oral epithelial dysplasia and to compare it with the WHO classification 2005. The accuracy of the new system for predicting malignant transformation was also assessed. Ninety-six consecutive oral epithelial dysplasia biopsies with known clinical outcomes were retrieved from the Oral Pathology archives. A pilot study was conducted on 28 cases to determine the process of classification. Four observers then reviewed the same set of H&E stained slides of 68 oral dysplastic lesions using the two grading systems blinded to the clinical outcomes. The overall inter-observer unweighted and weighted kappa agreements for the WHO grading system were Ks = 0.22 (95% CI: 0.11-0.35), Kw = 0.63 (95% CI: 0.42-0.78), respectively, versus K = 0.50 (95% CI: 0.35-0.67) for the new binary system. Interestingly, all pathologists showed satisfactory agreement on the distinction of mild dysplasia from severe dysplasia and from carcinoma in situ using the new WHO classification. However, assessment of moderate dysplasia remains problematic. The sensitivity and specificity of the new binary grading system for predicting malignant transformation in oral epithelial dysplasia were 85% and 80%, respectively and the accuracy was 82%. The new binary grading system complemented the WHO Classification 2005 and may have merit in helping clinicians to make critical clinical decisions particularly for the cases of moderate dysplasia. Histological grading of dysplasia using established criteria is a reproducible prognosticator in oral epithelial dysplasia. Furthermore, the present study showed that more consensus scoring on either the degree of dysplasia, assessment of risk or the presence of each morphological characteristic by a panel should be encouraged.

Expression of type 2 nitric oxide synthase and vascular endothelial growth factor in oral dysplasia

Purpose: The small molecule nitric oxide (NO), produced by a family of enzymes called NO synthase (NOS), has a diverse array of functions in both physiologic and pathologic states. Prolonged production of NO by the isoform NOS2 has been implicated in human cancer progression. NO has an important role in angiogenesis, being both an upstream signal and a downstream effector molecule to vascular endothelial growth factor (VEGF). The latter group of proteins are crucial for vascular endothelial cell proliferation and permeability. The expression of VEGF increases with cancer progression. Because angiogenesis is a prerequisite for the development of invasive cancer, this immunohistochemical study investigated the expression of NOS2 and VEGF in oral epithelial dysplasia. Materials and Methods: An immunohistochemical study was performed using monoclonal antibodies to NOS2 and VEGF on archival formalin-fixed, paraffin-embedded tissue of 33 cases of oral dysplasia. Results: A significant correlation was found between NOS2 and VEGF expression in oral dysplasia (P <.001). Expression of both NOS2 and VEGF also correlated with the severity of dysplasia (P <.001, P <.002). Conclusions: These findings may provide further understanding to the complex transformation of oral epithelial dysplasia to invasive carcinoma and the role of angiogenesis in this process. © 2002 American Association of Oral and Maxillofacial Surgeons J Oral Maxillofac Surg 60:1455-1460, 2002

Differential expression of heat shock protein 27 in normal oral mucosa, oral epithelial dysplasia and squamous cell carcinoma

The aim of this study was to evaluate HSP27 expression in fetal, normal and inflamed oral mucosal epithelium and in oral premalignant epithelial lesions and in their ensuing invasive cancers. In developing human oral epithelia, immunoreactions for HSP27 were moderately observed in suprabasal keratinocytes of palate and tongue. Normal oral epithelium had an intense suprabasal positivity. In inflamed oral mucosa, HSP27 staining was stronger in basal and suprabasal keratinocytes than in normal epithelium. Most oral premalignant lesions showed no (5 cases, 29%) or low (8 cases, 46.4%) staining. In OSCC both low and high HSP27 levels of expression were observed. HSP27 immunolabelling was down-regulated in poorly differentiated areas and up-regulated in highly differentiated ones. These findings indicated that HSP27 expression seems to protect cells from apoptosis during inflammation, while the down-regulation in dysplasia could impair the protective mechanism against mutagenesis induced by environmental factors and thus enhancing the transformation of oral epithelial dysplasia into OSCCs.