DNA ploidy in proliferative verrucous leukoplakia

Proliferative verrucous leukoplakia (PVL) is a clinicopathologically distinctive form of oral leukoplakia presenting with multifocal flat, nodular and verrucous lesions that progress inexorably to squamous carcinoma. The aims of this investigation were to describe the clinical and histopathological features of six cases of PVL and to determine whether lesional epithelium demonstrates DNA ploidy anomalies prior to malignant transformation. The clinical and pathological features of six patients were reviewed and all biopsy specimens were subjected to image-based DNA ploidy analysis. The female:male ratio was 5:1 and the average age on first biopsy was 66 years. Only one patient reported both tobacco smoking and alcohol intake. The most frequently affected sites were alveolar ridge and/or gingiva (6/6), buccal mucosa (3/6), palate (3/6), tongue (2/6), buccal sulcus (2/6), and lip (1/6). Three patients developed multiple primary carcinomas, either invasive or verrucous. A ploidy anomaly at any oral site would have predicted malignant transformation in four cases and probably in a fifth for whom DNA ploidy failed to meet diagnostic criteria but was suspicious of aneuploidy. The site of transformation was predicted by ploidy and histopathology for three carcinomas and a further carcinoma showed severe dysplasia and a suspicious ploidy result in adjacent tissue. Both conventional histopathology and DNA ploidy proved effective in predicting the site of transformation in this limited series.     

Increased expression of c-erbB-3 protein and proliferating cell nuclear antigen during development of verrucous carcinoma of the oral mucosa

BACKGROUND: The roles of c-erbB-3 gene and protein in the pathogenesis, progression, and outcome of oral carcinoma remain unknown. To determine whether c-erbB-3 expression could serve as an indicator of progression from premalignant to malignant transformation and of prognoses in patients with oral carcinoma, the authors examined the relation between the expression of c-erbB-3 protein and cell proliferation activity during the development of oral verrucous carcinoma (VC). METHODS: Immunohistochemical techniques were used to evaluate c-erbB-3 protein and proliferative cell nuclear antigen (PCNA). Sixty-one samples (36 patients) of verrucous hyperplasia (VH), VC, and well differentiated squamous cell carcinoma arising from verrucous carcinoma (V-WSCC) in the oral mucosa were examined. RESULTS: Normal human oral mucosa showed weak c-erbB-3 immunostaining, predominantly on the epithelial surface. In contrast, 7 (39%) of 18 VHs, 25 (84%) of 31 VCs, and all (100%) of 12 V-WSCCs demonstrated overexpression of c-erbB-3 protein with increased expression of PCNA in some premalignant epithelial cells and many tumor cells. This finding suggested the involvement of c-erbB-3 gene in the progression from VH to VC and V-WSCC. CONCLUSIONS: Overexpression of c-erbB-3 protein correlated with increased PCNA labeling index, indicating that c-erbB-3 may contribute to malignant transformation and tumor growth. Further, patients with high expression of both c-erbB-3 and PCNA had a poor outcome. Study results suggested that c-erbB-3 expression was an index of malignancy during progression from VH to VC and V-WSCC and might have been involved in the outcome of oral carcinoma patients.     

Verruciform xanthoma of the penis in a young male masquerading as squamous cell carcinoma: Case Report

Verruciform xanthoma (VX) is a relatively rare benign lesion of unknown etiology with a predilection for oral mucosa. Penis is an uncommon location and to date only 17 cases of VX of penis have been reported. The lesion assumes importance as it can be easily mistaken for verrucous carcinoma or squamous cell carcinoma. We present herein a case of penile VX in a 25-year-old man who was referred to our institute with a histopathologic diagnosis of squamous cell carcinoma reported elsewhere. On microscopy the lesion demonstrated florid verruciform hyperplasia of the epidermis with hyperkeratosis, parakeratosis and acanthosis. There was a dense infiltrate of foam cells in the papillary dermis (a hallmark of VX), which were positive for CD68 and negative for S-100 protein. We present this case to highlight the significance of recognizing VX in extra oral location. An awareness of this entity is crucial to prevent misdiagnosis and halt inappropriate therapeutic intervention. Keywords: penis, squamous cell carcinoma, verruciform xanthoma, verrucous carcinoma.     

Malignant transformation of proliferative verrucous leukoplakia to oral squamous cell carcinoma: a series of 55 cases

In this study, we compared clinical differences between patients with proliferative verrucous leukoplakia (PVL) who did and did not develop oral cancer. This was a retrospective study of 55 PVL cases. All patients were followed for at least 1year, with a mean follow-up of 7.53 (SD=4.18) years. The mean age of the patients was 61.69 (SD=11.76) years. There were 36 (65.5%) females and 19 (34.5%) males. Group 1 included the 28 PVL patients who did not develop oral squamous cell carcinoma (OSCC) during the period. Group 2 included the 27 PVL patients who developed at least one OSCC during the period. The latter group was subdivided into patients with PVL who developed one (Subgroup 2a; n=16) or more than one (Subgroup 2b; n=11) OSCC during follow-up. There was no statistical difference in age or oral location of the PVL between Groups 1 and 2. Females had a greater tendency to develop cancer than males (58.3% vs. 31.6%). In Group 2, only six (22.2%) were smokers. There was no difference between Subgroups 2a and b in terms of age, gender, and oral location of the PVL. Subgroup 2b included fewer smokers, had higher gingival involvement rate, and lower frequency of PVL on the tongue. The PVL patients who develop oral cancers were more commonly female and non-smokers. Those patients who developed more than one OSCC were most likely to develop lesions of the gingiva.     

Proliferative verrucous leukoplakia: unusual locations of oral squamous cell carcinomas, and field cancerization as shown by the appearance of multiple OSCCs

Proliferative verrucous leukoplakia (PVL) is an uncommon entity with a high tendency to develop oral squamous cell carcinomas (OSCCs). The objective of this study was to analyse the presence of various OSCCs in the same patient with PVL. We studied 19 patients with PVL who had developed at least one OSCC. We analysed how many of these developed more than one OSCC over a period between 24 and a maximum of 130 months, indicating the location of their OSCC, clinical type and the time lapse between the appearance of each of the different OSCCs in the same patient. Of the 19 patients, 10 presented more than one of these cancers, one of whom even went on to develop five different cancers. The most frequent location of OSCC was the gingiva and the palate; the least common was the tongue/floor of mouth. Ninety percent were women and 20% were smokers. The average time elapsed between the detection of the first tumour and the appearance of the second was 19.20 months (SD 13.41). Our patients with PVL developed a high frequency of OSCCs, on many occasions manifesting several cancers at different oral locations, thus demonstrating the field cancerization of this entity. The OSCC in PVL patients were at sites quite uncommonly affected in patients who develop OSCC in the absence of PVL.     

口腔癌、癌前病变及癌前状态中抑癌基因PTEN的表达及其临床意义的研究

目的研究PTEN蛋白在口腔鳞状细胞癌、癌前病变及癌前状态组织中的表达及其与口腔鳞状细胞癌的临床病理及预后的关系。了解PTEN蛋白表达在口腔癌发生、发展中的作用。方法免疫组织化学法检测10例正常口腔粘膜、14例口腔扁平苔癣、20例口腔白斑、30例癌旁组织及46例口腔鳞状细胞癌组织中PTEN蛋白的表达水平,并进行半定量分析。结果正常口腔粘膜、口腔扁平苔癣、口腔白斑、癌旁组织及鳞状细胞癌组织中PTEN蛋白表达总阳性率分别为70．00％（7／10）、57．14％（8／14）、45．00％（9／20）、73．3％（22／30）及19．57％（9／46）。口腔白斑、扁平苔癣及鳞状细胞癌组织中VrEN蛋白表达水平显著低于正常口腔粘膜（P〈0．05）。口腔鳞状细胞癌组织中PTEN蛋白表达水平显著低于患者自身癌旁组织（P〈0．01）。结论PTEN蛋白表达水平降低或缺失在口腔鳞状细胞癌的发生发展过程中可能起重要作用。免疫组织化学法检测PTEN蛋白的表达对口腔癌早期诊断和预后判断的价值有待进一步研究。     

Gene expression profiles of oral leukoplakia and carcinoma: genome-wide comparison analysis using oligonucleotide microarray technology

Although leukoplakia is the most common precancerous lesion of the oral cavity, its molecular biological properties are largely unknown. The aim of this study was to identify the genes responsible for its pathogenesis and malignant transformation using oligonucleotide microarray technology. The expression profiles of 8,800 genes in human oral leukoplakia (n=4) and oral squamous cell carcinoma (OSCC) (n=2) were analyzed using the Affymetrix GeneChip system and the results were confirmed with RT-PCR. Eight genes were up-regulated (>2.0-fold) and ten were down-regulated (<0.5-fold) in all leukoplakias analyzed with the GeneChip. In particular, loricrin and keratins displayed greater differences between normal tissue and leukoplakia. Some of the 18 alternatively expressed genes were markedly down-regulated in squamous cell carcinoma compared with leukoplakia. Our data suggested that gene abnormalities in cytoskeleton network components might be responsible for the development and progression of oral leukoplakia.     

Proliferative verrucous vs conventional leukoplakia: no significantly increased risk of HPV infection

Proliferative verrucous leukoplakia (PVL) is a very aggressive form of oral leukoplakia (OL) with high morbidity and mortality rates, hypothesised to be linked to HPV infection. This study aimed to determine the presence of HPV DNA in PVL in comparison with OL, and in relation to social-demographical variables (age, gender, smoking and drinking habits) in an Italian multi-centric hospital-based study. The study group consisted of 58 cases of PVL and 90 cases of OL as controls (47 homogeneous (H) and 43 non-homogeneous (non-H) form), both recruited from four Italian cohorts. HPV DNA was identified in exfoliated mucosal cells by nested PCR (nPCR) with MY09/MY11 and GP5+/GP6+ primer pairs and the HPV genotype determined by direct DNA sequencing. HPV DNA was found in 24.1% (14/58)of PVL and in 25.5% (23/90) of OL; there was thus no significant difference found between PVL and OL (both forms) for risk of HPV infection (OR=0.93; 95% IC:0.432-1.985). Similarly, in both groups of PVL and OL lesions, no statistic association was found between any demographical variable considered and HPV infection. HPV-18 was the most frequently detected genotype in all tissues, being found in 78.5% and 60.8% of HPV+ve PVL and OL, respectively. Other more rarely detected genotypes were HPV-16 (28.6% in PVL and 13% in OL), HPV-6 (17.4% in OL) and HPV-53 (8.8% in OL). PVL does not appear more likely to be associated to HPV infection than conventional OL lesions.     

Potency of Pfeiffer’s Crystallization to Analyze Oral Leukoplakia and Squamous Cell Carcinoma

Objective: Oral cancer usually has an early precancerous stage before its actual malignant transformation. Although there are various approaches to diagnose early stages of cancer, yet there is one less explored, cost effective and simple technique known as crystallization test. The aim of the study was to reaffirm the effectiveness of Pfeiffer’s crystallization test in screening oral leukoplakia and squamous cell carcinoma. Methods: Fifty oral leukoplakia, sixty five oral squamous cell carcinoma and sixty healthy individuals participated in crystallization test. Single blood drop was used to perform the test and obtained crystal patterns were analysed. Cross tabulation and Chi-Square test was performed to find the frequency and association between the groups. Kruskal-Wallis H test and Mann Whitney U test was applied comparing mean transverse form. Results: Sensitivity of crystallization test was 80% and 93.84% in oral leukoplakia and squamous cell carcinoma. Chi-Square analysis revealed highly significant transverse form between the study groups (p < 0.000). Conclusion: Crystallization test proves to be simple, reliable and minimal invasive diagnostic approach under strictly maintained physical conditions.     

Diagnostic Biomarkers in Oral Verrucous Carcinoma: A Systematic Review

Oral verrucous carcinoma (OVC), a low-grade variant of oral squamous cell carcinoma (OSCC), is most frequently seen in the oral cavity. No clear etiology has been found for this lesion, but human papilloma virus, chewing betel nuts, and ultraviolet radiation are suggested as probable causes. Differential diagnosis of OVC is challenging for oral pathologists. The aim of this study was to review the molecular-based approaches for differential diagnosis of OVC. An electronic search was conducted in Medline and Scopus from January 2004 to July 2015 limited to English language publications. Published papers on verrucous carcinoma (VC) were found according to the inclusion and exclusion criteria and analyzed qualitatively. Data extraction were performed according to PRISMA statement. A total of 423 articles were reviewed; out of which, 26 articles completely fulfilled the inclusion criteria. Most of the included studies investigated proliferative and apoptotic biomarkers such as p53 and Ki67. No definite conclusion was drawn for cytoskeletal biomarkers due to variability of factors and lack of significant expression. However, it seems that cytokeratin10 (CK 10) can be useful for differentiation of OVC and benign squamous lesions. Among cell surface and extracellular matrix biomarkers tissue biomarkers, matrix metalloproteinase (MMP)-2, ?9, CD31 and CD68 seem to be useful for differentiation of OVC and OSCC and glucose transporter-1 (GLUT-1) can help in differentiation of OVC from oral epithelial dysplasia. Differences among OVC, OSCC and normal epithelium in expression profiles of the investigated biomarkers help in their differential diagnosis; although, clinicohistopathological similarities among verrucous hyperplasia, noninvasive OVC and invasive well-differentiated OSCC make the diagnosis difficult. Further studies are required to better differentiate these oral lesions.     

Rapid progression from oral leukoplakia to carcinoma in an immunosuppressed liver transplant recipient

Immunosuppression used to avoid graft rejection in solid organ transplantation recipients leads to a variety of side-effects, and an increased rate of infections and de novo malignancies. Oral conditions usually associated with immunosuppressive drugs include fungal and viral infection, and lip lesions, but intra-oral carcinoma has not been reported as having a high incidence. This report deals with a male liver transplant recipient receiving FK506 (5 mg/day) and prednisone (20 mg/day) who was diagnosed with a homogeneous leukoplakia on the floor of the mouth 4 months after transplantation, and 4 months later with a squamous cell carcinoma growth at the site of this lesion. The rapid transformation of the lesion suggests that in patients who display oral premalignant conditions, immunosuppression must be considered as an important risk factor for oral cancer.     

Pathological and clinical analysis of 102 cases of lip carcinoma

102 cases (103 tumors) of lip carcinoma treated by surgery are analyzed. There were 37 (35.9%) verrucous carcinoma and 66 (64.1%) squamous cell carcinoma which included grade I 29 cases, grade II 21, grade III 14 and grade IV 2. Of the 66 squamous cell carcinomas, six lesions co-existing with verrucous carcinoma were derived from the latter. This fact demonstrated that anaplastic transformation of verrucous carcinoma could occur in patients without radiotherapy. This study suggests that the enlarged neck lymph nodes need not be dissected prophylactically for patients with squamous cell carcinoma grade I and grade II, and with verrucous carcinoma. The management of the cervical nodes still suffices if they enlarge after the primary lesion has been removed. But radical dissection of neck lymph nodes should be performed promptly for patients with squamous cell carcinoma grade III and grade IV.     

The results of CO2 laser surgery in patients with oral leukoplakia: a 25 year follow up

Oral leukoplakia is an important premalignant lesion of the oral mucosa. We treat this lesion prophylactically with CO2 laser evaporation. In the period from 1976 to 2001, a group of 200 patients with 282 oral leukoplakias were treated by CO2 laser evaporation. In a follow up period of 1-219 months (mean 52), 251 treated leukoplakias (89.0%) did not show a recurrence. Twenty eight (9.9%) local recurrences were observed in 5 to 168 months after treatment. Three (1.1%) squamous cell carcinoma, occurred in the treated area respectively 7, 17 and 19 month after CO2 laser evaporation. This large study with a long follow up shows that laser treatment is a good prophylactic treatment for oral leukoplakia.     

Comparison of Serum Fucose Levels in Leukoplakia and Oral Cancer Patients

Background: Tumor markers, designated as a broad group of substances produced by malignancies, could be in the form of biochemical substances, immunological substances, cell surface changes and genetic alterations. Cancer, a disorder of cellular behavior is characterized by alteration of serum glycoproteins. L-fucose, a hexose, which is the terminal sugar in most of the plasma glycoproteins, may be useful as a tumor marker for the detection, monitoring and prognostic assessment of malignancies. The aim of the study was to ascertain the role of serum fucose as a biomarker for early detection of oral cancer and to compare serum fucose levels in healthy controls, leukoplakia and oral cancer patients. Materials and Methods: The study included 60 (100.0%) subjects, who were grouped as 20 (33.3%) control subjects, 20 (33.3%) squamous cell carcinoma patients and 20 (33.3%) leukoplakia patients. Fucose estimation was done using UV-visible spectrophotometry based on the method as adopted by Winzler using cysteine reagent. The results were analyzed statistically using ANOVA with Bonferroni post hoc tests. Results: Results showed a high significance in serum fucose in oral squamous cell carcinoma (OSCC) and leukoplakia subjects compared to normal controls. There was a gradual increase in the values noted from control to leukoplakia and to squamous cell carcinoma. Conclusions: Estimation of serum fucose may be a reliable marker and can be used as an effective diagnostic biomarker in oral squamous cell carcinoma patients.     

High-resolution DNA flow cytometry in oral verrucous carcinoma

The classification of verrucous carcinoma as an entity unto itself or as a variant of well differentiated squamous cell carcinoma is controversial. To contribute new insights into the biological behavior of this rare tumor, we applied DNA flow cytometry to three node-negative verrucous carcinomas of the oral cavity. All tumors expressed a single aneuploid cell population. One of the patients experienced three courses with local recurrence. All secondary tumors retained the initially established aneuploid clone. The development of aneuploidy is thus a cytogenetic event common to both verrucous and squamous carcinoma of the oral cavity.     

Expression of alpha(v)beta6 integrin in oral leukoplakia

The distribution of alpha(v)beta6 integrin was examined in oral leukoplakia, lichen planus and squamous cell carcinomas using immunohistochemistry. Controls included oral mucosal wounds, chronically inflamed and normal oral mucosa. Integrins beta1, beta3, beta4, beta5, fibronectin and tenascin were also studied. The integrin alpha(v)beta6 was highly expressed throughout the whole lesion of 90% of the squamous cell carcinomas but was not present in any of the normal specimens. alpha(v)beta6 integrin was also expressed in 41% of the leukoplakia specimens, and 85% of the lichen planus samples, but in none of the tissues with inflammatory hyperplasia or chronic inflammation. The expression of beta1 integrins was localized in the basal layer, and that of the beta4 at the cell surface facing the basement membrane of all specimens. The integrins beta3 and beta5 were absent from all normal and leukoplakia specimens. Fibronectin and tenascin were present in the connective tissue underneath the epithelium of all the sections, and their expression was similar in both alpha(v)beta6-positive and alpha(v)beta6-negative tissues. A group of 28 leukoplakia patients were followed 1-4 years after first diagnosis. In this group, initially alpha(v)beta6 integrin-positive leukoplakia specimens had high tendency for disease progression while alpha(v)beta6-negative specimens did not progress. These results suggest that the expression of alpha(v)beta6 integrin could be associated in the malignant transformation of oral leukoplakias.     

Verrucous carcinoma in penile lichen sclerosus et atrophicus

A patient presented with a verrucous carcinoma within a lesion of penile lichen sclerosus et atrophicus. We report a case of lichen sclerosus and the specific squamous cell carcinoma variant, verrucous carcinoma, treated by Mohs surgery.     

Lectin binding to oral squamous carcinoma

Ten benign epithelial hyperplasias, ten invasive squamous carcinomas, and one each of verrucous carcinoma and carcinoma in situ of the oral mucosa were examined immunoenzymatically using biotinylated lectins and avidin-biotin-peroxidase complex to localize glycoconjugates in the epithelial cells. All of the lectins intercellularly bind to benign and malignant epithelial cells. Also, lectins bind to the cytoplasm of the basal cells and all layers of carcinoma in situ. Verrucous carcinoma shows a similar binding as does the benign epithelial hyperplasia. However, in invasive squamous carcinoma some nests and individual invasive cells show intense cytoplasmic binding and a loss of cell surface binding to lectins, especially with Con A, and may be a marker for invasive potential of squamous carcinoma.     

Verrucous carcinoma arising in ulcerative lichen planus of the soles

Verrucous carcinoma is a low-grade squamous cell carcinoma. Chronic inflammation is an important factor in its pathogenesis. This is the first case of plantar verrucous carcinoma arising in a lesion of ulcerative lichen planus. This paper also lends further support to the successful use of Mohs micrographic surgical technique as the treatment of choice for verrucous carcinoma.