Clinicopathologic characteristics of young patients with lip squamous cell carcinoma: a retrospective study.

BACKGROUND: This retrospective study investigates the clinicopathological features and outcomes of young and elderly patients diagnosed with lip squamous cell carcinoma (LSCC). MATERIAL AND METHODS: Data from LSCC patients from Dr. Luiz Antonio Hospital in Natal, Brazil (2000-2015) were analyzed, grouping individuals below 40 and above 60 years old. Demographics, lifestyle habits, clinicopathologic characteristics, and treatment outcomes were examined using descriptive statistics, Chi-square and Fisher's tests, and Kaplan-Meier survival analysis. RESULTS: A total of 47 patients was analyzed, being 20 younger and 27 older, finding significant age-related differences (p = < 0.0001). Although in both groups the tumor was more common in males, older patients had a higher rate of females (29.6%) (p=0.0358) and smoking (70.4%) (p = 0.0043) and underwent more modalities of treatments (p = 0.0027). There were no significant differences in the other analyzed clinicopathologic factors, and survival rates did not differ significantly, though younger patients showed slightly better survival metrics in univariate analysis. CONCLUSIONS: LSCC exhibits some distinct clinicopathological features across different age groups, with significant differences in treatment modalities and progression rates. Age-specific approaches may be required to optimize treatment outcomes.

Decreased Nuclear Immunoexpression of ING3 is a Frequent Event in Lip Carcinogenesis.

PURPOSE: Evaluate the immunohistochemical expression of the ING3 in actinic cheilitis and squamous cell carcinoma of the lower lip. METHODS: Forty-five specimens of actinic cheilitis and 48 specimens of squamous cell carcinoma of the lower lip were submitted to immunohistochemical detection of ING3. The protein expression in different cellular sublocations was compared between the two groups, and associations with the clinicopathological variables were analyzed. A significance level of 5% was adopted for all tests. RESULTS: Deaths were significantly more frequent in tumors with a high histopathological risk score (p < 0.05). In actinic cheilitis, significant differences were found in the nucleus-cytoplasmic expression of ING3 and expression restricted to the cytoplasm with binary histopathological grading (p < 0.05). In squamous cell carcinoma of the lower lip, there was no statistically significant difference when comparing ING3 expressions with clinical and morphological parameters (p > 0.05). Nucleo-cytoplasmic ING3 expression was significantly lower in squamous cell carcinoma of the lower lip when compared to actinic cheilitis (p < 0.05) and the expression restricted to the cytoplasm was significantly higher in squamous cell carcinoma of the lower lip (p < 0.05). CONCLUSION: The results of this study suggest that there is a marked decrease in the nuclear expression of ING3 as malignant progression occurs, indicating an impaired tumor suppressor function of this protein in actinic cheilitis and squamous cell carcinoma of the lower lip.

Assessment of the association of myofibroblasts and structural components of the extracellular matrix with histopathological parameters of actinic cheilitis and lower lip squamous cell carcinoma.

BACKGROUND: To evaluate the presence of myofibroblasts (MFs) in the development of lip carcinogenesis, through the correlation of clinical, histomorphometric and immunohistochemical parameters, in actinic cheilitis (ACs) and lower lip squamous cell carcinomas (LLSCCs). METHODS: Samples of ACs, LLSCCs, and control group (CG) were prepared by tissue microarray (TMA) for immunohistochemical TGF-ß, α-SMA, and Ki-67 and histochemical hematoxylin and eosin, picrosirius red, and verhoeff van gieson reactions. Clinical and microscopic data were associated using the Mann-Whitney, Kruskal-Wallis/Dunn, and Spearman correlation tests (SPSS, p < 0.05). RESULTS: ACs showed higher number of α-SMA+ MFs when compared to CG (p = 0.034), and these cells were associated with the vertical expansion of solar elastosis (SE) itself (p = 0.027). Areas of SE had lower deposits of collagen (p < 0.001), immunostaining for TGF-ß (p < 0.001), and higher density of elastic fibers (p < 0.05) when compared to areas without SE. A positive correlation was observed between high-risk epithelial dysplasia (ED) and the proximity of SE to the dysplastic epithelium (p = 0.027). LLSCCs showed a higher number of α-SMA+ MFs about CG (p = 0.034), as well as a reduction in the deposition of total collagen (p = 0.009) in relation to ACs and CG. There was also a negative correlation between the amount of α-SMA+ cells and the accumulation of total collagen (p = 0.041). Collagen and elastic density loss was higher in larger tumors (p = 0.045) with nodal invasion (p = 0.047). CONCLUSIONS: Our findings show the possible role of MFs, collagen fibers, and elastosis areas in the lip carcinogenesis process.

Relationship between tumor thickness and GATA3 immunoexpression in lip and tongue squamous cell carcinomas.

PURPOSE: Lower lip squamous cell carcinomas (LLSCCs) exhibit lower levels of aggressiveness, low relations with metastases and better prognosis when compared with intraoral squamous cell carcinomas. Differently from the oral tongue squamous cell carcinomas (OTSCCs) have a high tendency towards local invasion and lymph nodal dissemination. Our aim was to evaluate tumor thickness in cases of oral squamous cell carcinoma and correlate it with histological grade of malignancy and GATA3 immunoreactivity. METHODS: Sixty specimens (30 LLSCCs and 30 OTSCCs) were scanned and digitized for the subsequent measurement of tumor thickness, histopathological examination, and quantitative analysis of GATA3 in the parenchyma and stroma of the tumors. RESULTS: Tumor thickness was lower in LLSCC compared to OTSCCs. Immunohistochemical analysis of GATA3 in parenchyma, stroma and both compartments showed higher immunoreactivity in LLSCCs compared to OTSCCs. We observed a negative correlation between tumor thickness and GATA3 expression in parenchyma, stroma, and both compartments. Our results revealed the presence of GATA3 in all cases both in the parenchyma and in the stroma. Higher expression was more related to LLSCCs, which are known to be less aggressive tumors than OTSCCs. CONCLUSIONS: A greater tumor thickness was found in OTSCCs, which was correlated with lower expression of GATA3, suggesting that this protein is involved in the inhibition of proliferative, migratory, and invasive capacity.

Possible role of ALDH1 and CD44 in lip carcinogenesis.

BACKGROUND: Lip squamous cell carcinoma (LSCC) accounts for 12% of all head and neck cancers. It is caused by chronic exposure to ultraviolet light solar radiation and related to previous actinic cheilitis (AC). This study aimed to investigate the immunostaining of the putative cancer stem cells (CSC) markers ALDH1 and CD44 in AC (n=30) and LSCC (n=20). ALDH1 positivity was found to be statistically higher in LSCC than in AC lesions (p=0.0045), whilst CD44 expression was statistically higher in AC than in LSCC lesions (p=0.0155). ALDH1+ cells in AC lesions were associated with specific clinical features: a younger age (<60 years old), the female gender, white skin, not smoking or consuming alcohol, and a fast evolution, and not associated with the chronic exposure to UV radiation (p<0.0001). CD44 positivity was associated with patients who were male, feoderm, smoked, consumed alcohol, underwent occupational exposure to UV-radiation, and demonstrated lesions with log-time evolution (p<0.0001). ALDH1 + cells were associated with mild dysplasia using a system from the World Health Organization (WHO), and with a low risk of malignant transformation, according to the binary system (p<0.0001). CD44+ cells were also associated with moderated dysplasia, according to the WHO system. In LSCC, ALDH1 + cells were positively associated with patients who were older (≥ 60 years old), smokers, and with those who consumed alcohol (p<0.0001). CD44 + cells in LSCC were associated with older (≥ 60 years old) patients as well, but also with female patients, white skin, non-smokers, and individuals who did not consume alcohol (p<0.0001), all of whom showed distinct patterns in pre- and malignant lesions of both markers. Additionally, in LSCC, both ALDH1 and CD44 staining were associated with smaller tumor sizes (T1/T2; p<0.0001). In summary, although both ALDH1 and CD44 were associated with the presence of dysplasia in AC lesions, the present findings suggest that ALDH1 and CD44 may be activated by different etiopathogenic pathways, predominantly in distinct steps of oral carcinogenesis. CD44 would thus be more significantly related to the potentially malignant lesion, while ALDH1 would be closely linked to malignancy.

Clinicopathological Analysis of Actinic Cheilitis: A Systematic Review with Meta-analyses.

BACKGROUND: This systematic review aimed to conduct a complete investigation of the demographic aspects, clinicopathological features, degrees of epithelial dysplasia, and malignant transformation rate of actinic cheilitis. METHODS: The study was performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines and registered in the International Prospective Register of Systematic Reviews (CRD42020201254). A search without year and language restrictions was performed using PubMed/MEDLINE, Embase, Virtual Health Library, Scopus, Web of Science, and gray literature. Studies that provided information on patients with actinic cheilitis were included, excluding those with general information on other diseases or other types of cheilitis. Risk of bias was explored using the Joanna Briggs Institute tool. Narrative and quantitative data syntheses were performed using meta-analyses and subgroup analyses. Association tests were also performed. RESULTS: Thirteen studies (728 patients) were included. The most prevalent clinical signs were dryness (99%), blurred demarcation between the lip vermilion and skin (82%), scaling (69%), and atrophy (69%). Regarding epithelial dysplasia, a prevalence of mild dysplasia (34.2%), followed by moderate (27.5%), and severe (14.9%). The malignant transformation rate was 14%. Crusts, ulcerations, and erythematous areas were associated with lip carcinoma (p < 0.001), and scaling was associated with actinic cheilitis (p < 0.001). CONCLUSIONS: This study revealed several features of actinic cheilitis, providing an overview of the disease. It is suggested that new studies help develop policy guides for the standardization of clinical criteria, enabling more rigorous and homogeneous analysis of actinic cheilitis.

Epithelial-mesenchymal transition modulates lower lip carcinogenesis and promotes cancer progression.

OBJECTIVE: To investigate and compare the immunoexpression of E-cadherin, α-SMA, TGF-ß and Snail proteins between cases of actinic cheilitis (AC) and squamous cell carcinoma of the lower lip (LLSCC). STUDY DESIGN: E-cadherin, α-SMA, TGF-ß and Snail antibody immunostaining was analyzed semiquantitatively in 54 AC cases and in 49 LLSCCs. The cases were classified as low and high expression for analysis of the association with clinicopathological variables and overall survival (OS) and disease-free survival (DFS) rates. RESULTS: High expression of E-cadherin (cytoplasmic) (p = 0.001) and α-SMA (p < 0.001) was identified in LLSCCs, as well as low expression of TGF-ß in LLSCCs (p < 0.001) and high expression of Snail in AC cases (p = 0.006). Survival analysis revealed that high expression of α-SMA at the tumor invasion front, a network immunostaining pattern of this protein, and high expression of TGF-ß in tumor buds were significantly associated with poor OS (p < 0.05). There was a higher risk of death among LLSCC cases with high expression of α-SMA (HR = 5.90, p = 0.03). High expression of TGF-ß in tumor buds was significantly associated with poor DFS (p = 0.007) and with a higher risk of negative outcomes for DFS (HR = 4.44, p = 0.014). CONCLUSIONS: The present results suggest the potential involvement of dysregulation of proteins associated with epithelial-mesenchymal transition in the modulation of lip carcinogenesis and greater aggressiveness of LLSCC.

Assessment of the presence of interleukin 17+ macrophages and Th17 cells in situ in lip and oral tongue cancer.

Increasing clinical evidence indicates that Th17 cells may promote or inhibit tumor progression, however the exact role of these cells in Oral Squamous Cell Carcinoma (OSCCs) pathogenesis and progression remains unclear. Tumor associated macrophages are highly plastic phenotype cells which can differentiate as M1 or M2. The mechanism and cellular phenotype of IL-17 expressing macrophages are unknown. 40 cases of lip and 28 of tongue SCCs were submitted to immunohistochemical analysis, and histologically graded. In tongue cases TNM was analyzed. The number of IL-17+ T cells was higher in lip SCC (p = 0.028). IL-17+ macrophages was greater in tongue SCC (p = 0.014). There were more IL-17+ macrophages in the high-grade malignancy oral tongue SCCs (p = 0.016), yet there was no significant difference in the numbers of RORγt+ lymphocytes by histopathological or TNM analysis. This study provides evidence concerning IL-17's pleiotropic roles, being possibly dependent on its cellular sources in the tumor microenvironment.

Imunoexpressão das proteínas E-caderina, α-SMA, TGF-ß e Snail em queilites actínicas e carcinoma epidermoide de lábio inferior / Immunoexpression of E-cadherin, α-SMA, TGF-ß and Snail proteins in actinic cheilitis and squamous cell carcinoma of the lower lip

A transição epitélio-mesenquimal (TEM) é um processo biológico que vem sendo amplamente estudado em carcinoma epidermoide oral (CEO), porém, ainda raramente avaliado na carcinogênese labial. O objetivo deste estudo foi de investigar a imunoexpressão das proteínas E-caderina, α-SMA, TGF-ß e Snail em queilites actínicas (QA) diagnosticadas histopatologicamente como displasias epiteliais, e em carcinomas epidermoides de lábio inferior (CELI). A imunoexpressão de E-caderina, α-SMA, TGF-ß e Snail foi analisada de forma semiquantitativa em 54 casos de QAs e em 49 CELIs. Visando a associação dos achados imunoistoquímicos com as variáveis clinicopatológicas e taxas de sobrevida global (SG) e livre de doença (SLD), os casos foram classificados nas categorias baixa expressão e alta expressão. Não foram observadas associações estatisticamente significativas com nenhuma das proteínas analisadas com o grau de severidade das displasias epiteliais em QAs (p > 0,05). A análise imunoistoquímica em CELIs revelou que uma baixa expressão membranar da E-caderina no front tumoral estava significativamente associada a CELIs com ≥5 buds (p = 0,005) e alto escore tanto para o modelo BD (p = 0,009), quando para o proposto por Dourado et al. (2020) (p = 0,038), entretanto, não foram observadas associações significativas entre a imunoexpressão desta proteína com parâmetros clínicos (p > 0,05). Constatou-se ainda associações significativas entre baixa expressão de α-SMA com CELIs em estágios clínicos TNM I/II (p = 0,05), baixa profundidade de invasão (p = 0,006), < 5 buds (p = 0,027) e escore de risco baixo/intermediário, ao passo que a alta expressão desta proteína foi associada com o desfecho óbito (p = 0,009). Também foram encontradas associações significativas entre o padrão de imunomarcação em rede/em fuso de α-SMA com CELIs em estágios TNM III/IV (p = 0,031), com alta profundidade de invasão (p = 0,002), ≥5 buds (p = 0,027), alto escore de risco BD (p = 0,001) e desfecho óbito (p = 0,015). Em relação à proteína TGF-ß, percebeu-se associações estatisticamente significativas entre sua baixa expressão em buds tumorais com ausências de metástase linfonodal (p = 0,047) e de recidiva locorregional (p = 0,042), contudo, não foram observadas significâncias com nenhum dos parâmetros morfológicos (p > 0,05). A análise imunoistoquímica da proteína Snail não revelou nenhuma associação significativa com os parâmetros clinicopatológicos (p > 0,05). A análise de associação das imunoexpressões das proteínas entre as lesões estudadas revelou resultados significativos para uma alta expressão citoplasmática da E-caderina e CELIs (p = 0,001), alta expressão de α-SMA e CELIs (p < 0,001), baixa expressão de TGF-ß e CELIs (p < 0,001) e alta expressão de Snail e QAs (p = 0,006). A análise de sobrevida revelou que uma alta expressão de α-SMA no estroma do front tumoral (p = 0,013), padrão de imunomarcação em rede desta proteína (p = 0,046) e alta expressão de TGF-ß em buds tumorais (p = 0,043) estavam significativamente associadas à pior SG, além de que CELIs com alta expressão de α-SMA também apresentavam maior risco de óbito (HR = 5,90, p = 0,030). Uma alta expressão citoplasmática de TGF-ß em buds tumorais estava significativamente associada tanto à pior SLD (p = 0,007), quanto a maiores riscos de desfechos negativos para a SLD (HR = 4,44; p = 0,014). Os resultados do presente estudo sugerem que apesar da imunoexpressão das proteínas avaliadas não indicarem diferenças no grau de severidade histopatológica em QAs, desregulações dessas proteínas foram identificadas entre as lesões estudadas. Ademais, foi constatado que CELI com comportamento mais agressivo estava associado a baixa expressão da E-caderina membranar, alta expressão de αSMA e seu padrão em rede e baixa expressão de TGF-ß em buds tumorais (AU).

Epithelial-mesenchymal transition (EMT) is a biological process that has been widely studied in oral squamous cell carcinoma (SCC), however, it is still rarely evaluated in lip carcinogenesis. The aim of this study was to investigate the immunoexpression of E-cadherin, α-SMA, TGF-ß and Snail proteins in actinic cheilitis (AC) histopathologically diagnosed as epithelial dysplasia, and in lower lip squamous cell carcinoma (LLSCC). The immunoexpression of E-cadherin, α-SMA, TGF-ß and Snail was semiquantitatively analyzed in 54 cases of ACs and 49 LLSCCs. Aiming at association of immunohistochemical findings with clinicopathological variables and overall (OS) and disease-free (DFS) survival rates, cases were classified into low expression and high expression categories. There were no statistically significant associations with any of the proteins analyzed with the degree of severity of epithelial dysplasia in ACs (p > 0.05). Immunohistochemical analysis in LLSCCs revealed that low membrane expression of E-cadherin in tumor front was significantly associated with LLSCCs with ≥5 buds (p = 0.005) and high score for both BD model (p = 0.009) and the proposed model by Dourado et al. (2020) (p = 0.038), however, no significant associations were observed between immunoexpression of this protein and clinical parameters (p > 0.05). Significant associations were also found between low expression of α-SMA with LLSCCs in clinical stages TNM I/II (p = 0.05), low depth of invasion (p = 0.006), < 5 buds (p = 0.027) and score of low/intermediate risk, while high expression of this protein was associated with the outcome of death (p = 0.009). Significant associations were also found between α-SMA network/spindle immunostaining pattern with LLSCCs in TNM stages III/IV (p = 0.031), with high depth of invasion (p = 0.002), ≥5 buds (p = 0.027), high BD risk score (p = 0.001) and death outcome (p = 0.015). Regarding the TGF-ß protein, statistically significant associations were noticed between its low expression in tumor buds with absence of lymph node metastasis (p = 0.047) and locoregional recurrence (p = 0.042), however, no significance was observed with any of morphological parameters (p > 0.05). Immunohistochemical analysis of Snail protein did not reveal any significant association with clinicopathological parameters (p > 0.05). The association analysis of protein immunoexpression between the lesions studied revealed significant results for a high cytoplasmic expression of E-cadherin and LLSCCs (p = 0.001), high expression of α-SMA and LLSCCs (p < 0.001), low expression of TGF -ß and LLSCCs (p < 0.001) and high expression of Snail and ACs (p = 0.006). Survival analysis revealed a high expression of α-SMA in tumor front stroma (p = 0.013), a network immunostaining pattern of this protein (p = 0.046) and high expression of TGF-ß in tumor buds (p = 0.043) were significantly associated with worse OS, and LLSCCs with high expression of α-SMA also had a higher risk of death (HR = 5.90, p = 0.030). High cytoplasmic expression of TGF-ß in tumor buds was significantly associated with both worse DFS (p = 0.007) and higher risks of negative outcomes for DFS (HR = 4.44; p = 0.014). The results of present study suggest that although the immunoexpression of evaluated proteins does not indicate differences in degree of histopathological severity in ACs, dysregulations of these proteins were identified among the lesions studied. Furthermore, it was found that LLSCC with more aggressive behavior was associated with low expression of membrane E-cadherin, high expression of α-SMA and its network pattern, and low expression of TGF-ß in tumor buds (AU).

Implication of the New AJCC pT Classification of SCC of the Lip Comparing With Other Oral Subsites.

OBJECTIVE: To determine the implication of the new AJCC staging system for pT classification in a cohort of patients with SCC of the lip mucosa and compare it to other oral cavity sites. METHODS: Retrospective cohort of 744 patients treated between 2002 and 2017, by the Head and Neck Surgery Department of the University of Sao Paulo. RESULTS: Of 95 lip patients, 42 had pT upstage (58.1% of pT1 to pT2-3 and 50% of pT2 to pT3). Similar DFS/OS observed for those pT1 maintained or upstaged to pT2-3, pT2 patients upstaged to pT3 presented worse OS (49.4% versus 92.3%, P = .032). The comparison between lip and other mouth topographies, denoted better prognosis for pT1-2, but not for pT3-4a. Lip tumors had lower DOI, rates of perineural/angiolymphatic invasion, nodal metastasis, recurrence, and death. CONCLUSION: The inclusion of DOI to the new pT classification better stratifies patients with SCC of the lip mucosa upstaged to pT3 by assessing inferior OS. LEVEL OF EVIDENCE: 3 Laryngoscope, 131:E2770-E2776, 2021.

Demographic and Clinicopathologic Features of Actinic Cheilitis and Lip Squamous Cell Carcinoma: a Brazilian Multicentre Study.

Data on the occurrence and clinicopathological characteristics of actinic cheilitis (AC) and lip squamous cell carcinoma (LSCC) are well studied; however, they are based on studies limited to a single centre. Herein, we described the frequency of AC and LSCC submitted to microscopic examination from representative geographic regions of Brazil. A retrospective multicentre study was performed on biopsies obtained from 1953 to 2018 at 10 Brazilian oral and maxillofacial pathology centres. A total of 198,709 biopsy specimens were surveyed. Sociodemographic data and clinicopathologic characteristics were analysed. A total of 2017 cases of ACs (1.0%) and 850 cases of LSCCs (0.4%) were recorded. A strong fair-skinned (> 87%) male (> 70%) predilection was observed in both conditions. The mean age was 54.8 ± 18.7 for individuals with AC and 57.8 ± 19.0 for individuals with LSCC. The most commonly affected site was the lower lip (> 90%). This is a large multicentre study of AC and LSCC from Brazil. The frequency and clinicopathological features of AC and LSCC were similar to those described worldwide. This study provides robust and representative epidemiological data of these conditions for the scientific community.

Immunoexpression of human leukocyte antigen-DR in actinic cheilitis and lower lip squamous cell carcinoma.

The aim of this study was to evaluate the immunoexpression of human leukocyte antigen-DR (HLA-DR) in actinic cheilitis (AC) and lower lip squamous cell carcinoma (LLSCC), and to correlate the findings with clinical (tumor size/extent, regional lymph node metastasis, and clinical stage) and histopathological (grade of epithelial dysplasia and inflammatory infiltrate for AC and histopathological grade of malignancy for LLSCC) parameters. Twenty-four AC and 48 LLSCC cases (24 with regional nodal metastasis and 24 without regional nodal metastasis) were selected. The scores of immunopositive cells for HLA-DR in the epithelial component of the lesions were assessed and the results were analyzed statistically using the nonparametric Mann-Whitney test. Epithelial expression of HLA-DR was observed in only five (20.8%) cases of AC (two low-grade and three high-grade lesions), with a very low median score of immunopositivity. By contrast, expression of HLA-DR was found in most LLSCC (97.9%), with a relatively high median score of positive cells. The score of HLA-DR-positive cells tended to be higher in tumors with regional lymph node metastasis, tumors in advanced clinical stages, and low-grade tumors, but the difference was not statistically significant (p > 0.05). In addition, there was a tendency towards higher expression of HLA-DR in highly/moderately keratinized tumors, and tumors with little/moderate nuclear pleomorphism (p > 0.05). The results suggest a potential role of HLA-DR in lip carcinogenesis, particularly in the development and progression of LLSCC. The expression of this protein can be related to the degree of cell differentiation in these tumors.

Predictive factors for late cervical metastasis in stage I and II squamous cell carcinoma of the lip.

PURPOSE: Many authors have described clinicopathologic parameters as factors related to cervical lymph node metastasis development in CN0 stage lip cancer. However, predictive factors for occult lymph node metastasis and criteria for elective neck dissection, especially for early tumour, remain undefined. METHODS: A multi-institutional study with 193 consecutive patients with early lip SCC treated from January 1990 to March 2006 was carried out retrospectively to determine factors predicting occult metastasis. RESULTS: The overall late LNM rate was 13% (25/193). In the multivariate logistic regression study, tumour size and pattern of tumour invasion were factors related to the occurrence of late LNM with rates of sensitivity, specifity and accuracy for occult LNM prediction of 50%, 89.5% and 87%, respectively. CONCLUSION: Our results indicate that patients with stage I and II SCC of the lip with tumour size greater than 18 mm and more aggressive pattern of invasion must be considered a high-risk group for LNM and an END should be performed.

Immunohistochemical evaluation of HLA-G and FoxP3+ T regulatory cells in oral cavity and lower lip squamous cell carcinomas.

Human Leukocyte Antigen G (HLA-G) is a molecule involved in the tumor immunosuppression and also in the generation of regulatory T (Treg) cells, thus leading to evasion to the immune system host, and consequently, contributing to tumor progression in several cancers. The aim of this study was to evaluate the immunoexpression of HLA-G by tumor cells and FoxP3+ Treg cells in 25 oral tongue squamous cell carcinomas (SCCs) and 25 lower lip SCCs and analyze their relationship with clinical parameters. HLA-G expression was higher in oral tongue SCCs than in lower lip SCCs. In oral tongue SCCs and lower lip SCCs, no association between HLA-G expression and clinical parameters (tumor size, lymph node status, distant metastasis, and clinical stage) was verified (P>0.05). FoxP3+ Treg cells were detected along the tumor invasive front in all cases of oral tongue and lower lip SCCs. In oral tongue SCC cases, the number of Treg cells tended to be higher in smaller tumors, tumors without regional lymph node metastasis, and tumors in early clinical stages, but the difference was not statistically significant (P>0.05). A significant positive correlation was found between the expression of HLA-G by neoplastic cells and Treg cells in lower lip SCCs (p = 0.008). Our findings suggest the involvement of HLA-G and Treg cells in the modulation of immune responses in oral tongue and lower lip SCCs. This interaction between HLA-G and Treg cells may represent an evasion mechanism in these malignancies.

The stem cell markers expression CD44v6 and podoplanin in lip cancer: clinical significance.

This study aimed to analyze the immunoexpression of cancer stem cell markers, CD44v6, and podoplanin in 91 patients with lip squamous cell carcinomas (LSCC). The immunostaining of podoplanin and CD44v6 was evaluated in ten high-power fields (× 400 magnification) at the invasive front of LSCC, using a semi-quantitative score method. Chi-square test or Fisher's exact test was used to verify the association of podoplanin and CD44v6 expressions with clinicopathologic variables. Spearman's correlation test was used to analyze the correlation between the two antibodies in lip cancer. Disease-free survival probabilities in 5 and 10 years were estimated according to the Kaplan-Meier method and compared using the log-rank test. The independent effects of the significant variables were analyzed by Cox proportional hazards regression model. A strong podoplanin expression was observed in the membrane and cytoplasm of most lip tumor cells, and this was inversely associated with locoregional recurrence (p = 0.028) and with histopathological grade of malignancy (p = 0.026). Additionally, CD44v6 immunostaining was strongly expressed in the membrane of tumor cells in 95.4% of the LSCC. Patients with strong membranous (p = 0.016) or strong cytoplasmic (p = 0.030) podoplanin-positive tumors resulted in significantly better disease-free survival than those who had podoplanin weak/negative tumors, confirming podoplanin expression as a favorable independent prognostic factor. Podoplanin and CD44v6 were strongly expressed by tumor cells and podoplanin immunoexpression can help to determine lip cancer patients with lower risk for disease recurrence.

Twist and E-cadherin deregulation might predict poor prognosis in lower lip squamous cell carcinoma.

OBJECTIVE: The aim of this study was to evaluate the expression of Twist and E-cadherin in lower lip squamous cell carcinoma (LLSCC) and their association with clinicopathologic parameters. STUDY DESIGN: Fifty-nine cases of LLSCC were analyzed by applying immunohistochemistry techniques in a semiquantitative manner. The systems proposed by Bryne etal., Brandwein-Gensler etal., and Almangush etal. were applied for analysis of the histopathologic malignancy grading system. RESULTS: Higher E-cadherin expression (general and membrane) was observed in cases presenting with disease-free survival after 5years of follow-up (P < .05). Higher Twist expression was observed in lesions classified as being in advanced stages, displaying recurrence, and having a high degree of malignancy. A significant negative correlation was detected between cytoplasmic Twist expression and membrane E-cadherin expression (P = .028). A statistically significant relationship was detected between high total Twist expression in tumors classified as high risk by Brandwein-Gensler etal., and no significant difference was observed among total, membrane, and cytoplasmic E-cadherin expressions in LLSCC cases and the 3 applied grading systems (P > .05). CONCLUSIONS: The results of the present study suggest the potential involvement of Twist and E-cadherin in the modulation of events related to worse prognoses in LLSCC cases.

Immune response and evasion mechanisms in lip carcinogenesis: An immunohistochemical study.

OBJECTIVES: Programmed death ligand-1 (PD-L1) and human leukocyte antigen-G (HLA-G) are considered immune checkpoint molecules that inhibit T-cell effectiveness, contributing to tumor immune escape. This study investigated PD-L1, HLA-G, CD8, and granzyme B (GrB) expression at different stages of lip carcinogenesis. DESIGN AND RESULTS: Forty cases of lip squamous cell carcinoma (LSCC), 55 actinic cheilitis (AC), and 10 healthy lip mucosa (HLM) were submitted to immunohistochemistry. Semiquantitative (PD-L1, HLA-G), and quantitative (CD8, GrB) analysis were performed. PD-L1 and HLA-G expression in neoplastic cells/keratinocytes and stroma/connective tissue was significantly higher in LSCC and AC, compared to HLM (p<0.05). PD-L1 was not associated with clinicopathological features of the lesions. HLA-G expression by malignant cells was significantly higher in LSCCs with distant metastasis (p = 0.041).CD8+ and GrB+ cell numbers progressively increased from HLMs to LSCC, with AC exhibiting intermediate numbers (p<0.01). Most LSCCs showed coexistence of PD-L1+ and CD8+ cells (72.5%). PD-L1 was directly correlated to CD8+ and GrB+ lymphocytic infiltration in LSCCs (p<0.05). Low cytotoxic immune response was associated with lymph node metastasis in LSCC (p<0.05). CONCLUSIONS: PD-L1 and HLA-G-mediated immune evasion mechanisms are likely to occur from early pre-malignant to advanced malignant stages of lip carcinogenesis, which might provide a rationale for therapeutic blockade of these pathways. PD-L1 expression in LSCCs was correlated with the cytotoxic markers, suggesting that PD-L1 may appear as an escape mechanism in response to an active antitumor response.

Estudo da imunoexpressão do reg-gamma e da beta catenina em queilites actínicas e carcinomas de células escamosas de lábio inferior / Study of immunoexpression of reg-gamma and beta-catenin in actinic cheilitis and lower lip squamous cell carcinoma

A queilite actínica (QA) é uma lesão potencialmente maligna que ocorre principalmente em homens leucodermas com histórico de exposição crônica ao sol. Atualmente, não é possível predizer quais os casos de QA progredirão para o Carcinoma de células escamosas (CCE), portanto alguns marcadores biomoleculares têm sido alvo de pesquisas. A ß-catenina é uma proteína multifuncional que está envolvida nos processos de adesão célula-célula. A alteração do complexo caderina-catenina tem sido demonstrada no CCE e correlacionada com a invasão tumoral, metástase e com pior prognóstico dos pacientes. O REGγ é um ativador de proteassoma que pode promover a degradação de múltiplas proteínas incluindo p53 e MDM2. Estudos mostram que o REGγ está superexpresso em numerosos tipos de câncer, sugerindo que a superexpressão do REGγ está envolvida na progressão do câncer. O objetivo deste estudo foi analisar a expressão imuno- histoquímica da ß-catenina e do REGγ em casos de QA e Carcinoma de células escamosas de lábio inferior (CCELI), comparando os achados imunohistoquímicos com os dados clínicopatológicos, afim de averiguar se há uma correlação com a progressão tumoral e se as mesmas atuam de forma sinérgica nesse processo. A imunoexpressão de ß-catenina e REGγ foi analisada semi-quantativamente em 30 casos de QA e 30 casos de CCELI de acordo com os escores: 0 (sem marcação); 1 (1-25% de células positivas); 2 (26-50% de células positivas); 3 (51-75% de células positivas); 4 (> 75% células positivas). Para a análise estatística, foram realizados os testes de Mann-Whitney e de Spearman (p < 0,05). Tantos as QAs quanto os CCELIs expressaram a proteína ß-catenina, sendo evidenciado um aumento da expressão citoplasmática e nuclear nos casos de Displasias moderadas e severas. Nos CCELIs a imunoexpressão de ß-catenina membranar foi maior nos casos de baixo grau de malignidade. Tantos as QAs quanto os CCELIs expressaram a proteína REG-γ porém não verificamos significância estatística entre a sua expressão e o grau displasia epitelial, bem como, entre a imunoexpressão do REG-γ e os parâmetros clinicopatológicos analisados nos CCELIs. Os resultados do presente estudo sugerem que a superexpressão de REG-γ e a redução na expressão membranar de ß-catenina podem ser eventos importantes na carcinogênese labial. No entanto, acreditamos que esta proteína esteja envolvida no processo da carcinogênese oral. Nesse processo, correlacionando a expressão imuno-histoquímica da ß-catenina com a expressão do REG-γ, não resultados estatisticamente significativos, sugerimos então que a expressão de ßcatenina pode não ser influenciada diretamente pelos níveis de expressão de REGγ (AU).

Actinic cheilitis (QA) is a potentially malignant lesion that occurs mainly in men with light skin and a history of chronic sun exposure. Currently, it is not possible to predict which cases of QA will progress to Squamous Cell Carcinoma (SCC), so some biomolecular markers have been researched. Β-catenin is a multifunctional protein that is involved in cell-cell adhesion processes. The alteration of the cadherin-catenin complex has been demonstrated in SCC and correlated with tumor invasion, metastasis and worse prognosis of patients. REGγ is a proteasome activator that can promote the degradation of multiple proteins including p53 and MDM2. Studies show that REGγ is overexpressed in numerous cancers, suggesting that REGγ overexpression is involved in cancer progression. The aim of this study was to analyze the immunohistochemical expression of ß-catenin and REGγ in cases of QA and lower lip squamous cell carcinoma (CCELI), comparing the immunohistochemical findings with the clinical and pathological data, in order to verify if there is any a correlation with tumor progression and whether they act synergistically in this process. Β-catenin and REGγ immunoexpression was analyzed semi-quantitatively in 30 cases of QA and 30 cases of CCELI according to the scores: 0 (no labeling); 1 (1-25% positive cells); 2 (26-50% positive cells); 3 (51-75% positive cells); 4 (> 75% positive cells). For statistical analysis, Mann-Whitney and Spearman tests were performed (p <0.05). Both QAs and CCELIs expressed the ß-catenin protein, showing an increase in cytoplasmic and nuclear expression in cases of moderate and severe dysplasias. In CCELIs, membrane ß-catenin immunoexpression was higher in cases of low grade malignancy. Both QAs and CCELIs expressed the REG-γ protein but no statistical significance between its expression and the degree of epithelial dysplasia was found, as well as between the immunoexpression of REG-γ and the clinicopathological parameters analyzed in the CCELIs. The results of the present study suggest that REG-γ overexpression and reduction in membrane expression of ß-catenin may be important events in lip carcinogenesis. However, we believe that this protein is involved in the process of oral carcinogenesis. In this process, correlating the immunohistochemical expression of ß-catenin with the expression of REG-γ, as we did not obtain statistically significant results, we suggest that ß-catenin expression may not be directly influenced by the levels of REGγ expression (AU).