Influence of p16 Protein Immunostaining on Histopathological Features of Pleomorphic Adenoma and Carcinoma ex- Pleomorphic Adenoma.

The objective of this study was to evaluate the role of p16 in histologic characteristics and transition of Pleomorphic Adenoma (PA) to Carcinoma ex-PA (CxPA). So, 60 PA and 4 CxPA were histologic reviewed based on microscopic characteristics proposed by Hellquist, Triantafyllou and Dulguerov (PA) and Morais, Antony and Toluie (CxPA). Immunostaining for p16 was associated in different parenchyma and stroma of both tumors and Fisher's/chi-square tests and Mann-Whitney test were performed (SPSS v20.0, p<0.05). In PA the periductal cells were predominantly p16- and that ductal and myoepithelial cells showed a significant increase in p16+ cells (p<0.001). In CxPA, none of the cases showed p16+ in periductal cells, most parotid cases showed p16+ in ductal cells, and one case of parotid and the submandibular case showed mild immunostaining for myoepithelial cells. There was a small reduction in p16+ in CxPA compared to PA (p=0.537), but in both tumors there was less p16+ cells in solid stroma than other (p<0.001). The p16+ cases of PA had a higher capsular thickness (p=0.047). So, the loss of p16 immunostaining does not seem to be associated with the transition from PA to CxPA, but in both tumors the loss of p16+ cells are related to microscopic aggressiveness.

P16 gene promoter methylation is associated with oncogenesis and progression of gastric carcinomas: A systematic review and meta-analysis.

INTRODUCTION: Gastric cancer is the fourth leading cause of neoplastic morbidity worldwide, and its pathogenesis has been related to genetic and epigenetic alterations in cell cycle regulatory genes, such as p16. OBJECTIVES: This systematic review was developed to investigate the association of the p16 gene promoter methylation with oncogenesis and the progression of gastric carcinomas. METHODOLOGY: MEDLINE and Scopus databases were searched for relevant subsidiary studies, with the publication until July 2021 and without language restriction. The complete protocol was registered in the PROSPERO platform under the identification 308218. The Newcastle-Ottawa scale (NOS) was used to assess the methodological quality of the included manuscripts. The meta-analysis was conducted using RevMan 5.4 ® software. The random effects model was used, Odds Ratio (OR) was calculated with 95% confidence intervals (95% CI). Heterogeneity and inferential significance were measured. RESULTS: 48 articles were aggregated in the qualitative synthesis and 47 in the meta-analysis, totaling 6599 gastric specimens evaluated. Associations of p16 methylation with the following outcomes were observed: gastric oncogenesis (p < 0.00001); intestinal metaplasia (p = 0.002); poor histological differentiation (p = 0.03); local invasion (p = 0.001); lymph node dissemination (p = 0.03); more advanced TNM staging (p = 0.01); and Epstein Barr virus infection (p < 0.00001). In contrast, no association of p16 methylation was found with Lauren's histological classification (p = 0.62); distant metastasis (p = 0.71); or Helicobacter pylori infection (p = 0.79). CONCLUSIONS: the findings described provide empirical evidence for the categorization of p16 methylation as a substantial biomolecular step in gastric carcinogenesis, and reveal a crucial role of Epstein Barr virus in triggering this epigenetic alteration.

Expression and clinical significance of estrogen receptor, progesterone receptor, p53 and p16 protein in endometrial carcinoma / 中国基层医药

Objective:To investigate the expression and clinical significance of estrogen receptor (ER), progesterone receptor (PR), p53 and p16 protein in endometrial carcinoma.Methods:The endometrial tissue of 57 patients with endometrial carcinoma who received surgery in The First People's Hospital of Chuzhou between January 2017 and May 2021 was harvested as the study group. The normal endometrial tissue of 30 patients with endometrial hyperplasia was selected as the control group. Envision immunohistochemical staining was performed to determine the expression of ER, PR, p53 and p16 protein in endometrial tissue and analyze their expression with clinical pathological characteristics.Results:ER, PR, p16 protein expression rates in the endometrial tissue in the study group were 70.2%, 61.4%, 38.6%, respectively, which were significantly lower than 90.0%, 86.7%, 93.3% in the control group ( χ2 = 4.36, 5.98, 24.09, all P < 0.05). p53 expression rate in the endometrial tissue was significantly higher in the study group than that in the control group (52.6% vs. 13.3%, χ2 = 12.75, P < 0.001). ER and PR expression were significantly different between endometrial carcinoma patients with lymph node metastasis and those without and among those with different histological grades and those at different pathological stages (all P < 0.05). There was no significant difference in p53 protein expression among patients with different pathological stages of endometrial carcinoma, between patients who suffered endometrial carcinoma at different ages, and between patients with different degrees of myometrial invasion (all P > 0.05). p16 protein expression rate differed among patients with different pathological stages of endometrial carcinoma, among those with different histological grades and between patients with different degrees of myometrial invasion (all P < 0.05). There was no significant difference in p16 protein expression rate between endometrial carcinoma patients with lymph node metastasis and those without ( P > 0.05). Conclusion:Abnormal expressions of ER, PR, p53 and p16 protein in endometrial tissue may be related to the occurrence, development and transformation of the disease. Combined detection of ER, PR, p53 and p16 protein is helpful for the clinical diagnosis, treatment and prognosis assessment of endometrial carcinoma.

Presença do papilomavirus humano tipo 16 e expressão gênica da proteína p16ink4a e oncoproteína e7 no carcinoma colorretal / Hpv16 and expression of protein p16ink4a and e7 oncoprotein in colorectal carcinoma

RESUMO - INTRODUÇÃO: O papilomavírus humano (HPV) é agente das doenças sexualmente transmissíveis de maior prevalência no mundo que estão associadas ao câncer do colo do útero e canal anal. A ação do HPV na carcinogênese colorretal não está ainda estabelecida. OBJETIVO: Estudar a eventual correlação entre a presença do HPV tipo 16 e a expressão gênica da proteína p16INK4a e da oncoproteína E7 de HPV e de seus níveis no tecido do carcinoma colorretal. METODOS: Estudo retrospectivo caso-controle de 79 doentes com carcinoma colorretal divididos em dois grupos: HPV presente e HPV ausente. Foi realizada reação em cadeia da polimerase (PCR), além da hibridização do tipo dot blot para o HPV 16 e o HPV 18 Amostras do tecido colorretal também foram submetidas ao estudo imuno-histoquimico para avaliar o nível tecidual das proteínas E7 e p16INK4a. RESULTADOS: O HPV foi identificado em 36 (45,6%) casos. Não houve diferença significante entre os grupos quanto ao sexo (p=0,056), idade (p=0,1), localização cólica e/ou retal (0,098) e presença do HPV. A expressão gênica da oncoproteína E7 de HPV estava presente em 3,12% dos casos (p=0,9) e a expressão da proteína p16INK4a foi observada em 46,3% (p=0,27) dos indivíduos com detecção do HPV. CONCLUSÃO: A expressão gênica e os níveis teciduais da oncoproteína E7 e da proteína p16INK4a encontrados nos pacientes positivos para o HPV sugerem a ausência de atividade oncogênica do HPV tipo 16 no carcinoma colorretal.

ABSTRACT - BACKGROUND: Human papillomavirus (HPV) is the agent of the most prevalent sexually transmitted diseases in the world associated with cervix and anal canal cancer. The action of HPV on colorectal carcinogenesis is not yet established. OBJECTIVE: This research aimed to study the possible correlation between the presence of HPV16 and the gene expression of p16INK4a protein and HPV E7 oncoprotein and their levels in colorectal carcinoma tissue. METHODS: A retrospective case-control study of 79 patients with colorectal carcinoma was divided into two groups: HPV-positive and HPV-negative. The polymerase chain reaction was performed, in addition to dot-blot hybridization for HPV16 and HPV18. Colorectal tissue samples were also subjected to immunohistochemical study to assess the tissue level of E7 and p16INK4a proteins. RESULTS: HPV was identified in 36 (45.6%) cases. There was no significant difference between groups regarding gender (p=0.056), age (p=0.1), colic and/or rectal location (0.098), and presence of HPV. Gene expression of HPV E7 oncoprotein was present in 3.12% of cases (p=0.9), and p16INK4a protein expression was observed in 46.3% (p=0.27) of those selected with HPV detection. CONCLUSION: Gene expression and tissue levels of E7 oncoprotein and p16INK4a protein found in HPV-positive patients suggest the absence of HPV16 oncogenic activity in colorectal carcinoma.

Is there correlation between human papillomavirus (hpv) and esophageal epidermoid carcinoma? / Há correlação entre o papilomavírus humano (hpv) e o carcinoma epidermoide de esôfago?

ABSTRACT Background: Currently, persistent human papillomavirus (HPV) infection has been related in some geographic regions as a risk factor for esophageal squamous cell carcinoma. It results in the immunoexpression of the p16 protein, which has been used as marker of the oncogenic lineage by this etiological agent. Aim: To correlate epidemiological aspects of esophageal squamous cell carcinoma with the prevalence of HPV infection. Methods: Fifty-eight cases were analyzed and submitted to histopathological and immunohistochemical analysis by p16. Results: Of the 58 cases evaluated, 40 were men and 18 women, with a mean age of 63.2 years. p16 immunoexpression was positive in 46.55%. Conclusion: The prevalence of HPV infection is high in esophageal squamous cell carcinoma presenting in almost half of the cases (46.55%), without gender differentiation.

RESUMO Racional: Atualmente a infecção persistente pelo papilomavírus humano (HPV) tem sido relacionada em algumas regiões geográficas como fator de risco para o carcinoma epidermoide do esôfago. Ela resulta na imunoexpressão da proteína p16, que tem sido utilizada como marcadora da linhagem oncogênica por este agente etiológico. Objetivo: Correlacionar aspectos epidemiológicos do carcinoma epidermoide do esôfago com a prevalência de infecção pelo HPV. Métodos: Foram analisados 58 casos buscando-se perfil epidemiológico dos pacientes, com suas peças submetidas à análise histopatológica e imunoistoquímica pelo p16. Resultado: Dos 58 casos avaliados, 40 eram homens e 18 mulheres, com idade média de 63,2 anos. A imunoexpressão pelo p16 foi de 46,55%. Conclusão: A prevalência de infecção pelo HPV é alta no carcinoma epidermoide de esôfago apresentando-se em quase a metade dos casos (46,55%), sem diferenciação de idade quanto aos gêneros.

Prognostic impact of p16 and p53 gene expressions in stage 1a epithelial ovarian cancer.

OBJECTIVE: Epithelial ovarian cancer (EOC) is rarely detected at stage 1a. Most of the patients have a good prognosis and there are limited factors that affect their survival. In the present study, we evaluated the p16 and p53 gene expressions of stage 1a EOC patients. Prognostic effects of these gene expressions, as well as those of other factors on short term survival were analyzed. METHODS: Our study included 29 patients. The specimens of the ovary with cancer were stained for p16 and p53. Gene expressions and other prognostic factors were evaluated. RESULTS: The median age of the patients was 51 years (27-84). The mean numbers of dissected pelvic and paraaortic lymph nodes were 27 and 12, respectively. The mean follow-up time was 33.7±18.9 months. During this period, recurrence occurred in two patients. One of the patients had grade 2 mucinous carcinoma and died of the disease at month 12 after the recurrence occurred at month 7. The second patient had clear cell carcinoma and recurrence occurred at month 34. p16 and p53 gene expressions or other factors were not associated with overall survival (OS) or disease-free survival in the short term. The lower p16 positivity rate in the non-clear cell group was found to be statistically significant (P=0.003). Both p53 and p16 positivity rates were higher in the high-grade carcinoma. CONCLUSION: The levels of none of the common prognostic factors, including those of p16 and p53 gene expression, were associated with the progression-free survival or OS of stage 1a in the short term. Appropriate surgical staging and non-omission of subclinical metastases seem to be of central importance.

Clinical value of P16 and Ki-67 expressions in cervical biopsy for predicting early cervical cancer in patients with high-risk HPV infection / 中国基层医药

Objective@#To explore the positive expressions of P16 and Ki-67 proteins in cervical biopsy lesions of patients with high-risk HPV infection and abnormal detection of liquid-based thin-layer cytology test(TCT), and the clinical value in predicting occurrence of early cervical cancer.@*Methods@#A total of 120 patients with high-risk HPV infection(16 and 18 subtypes positive) and abnormal TCT in the Traditional Chinese Medicine Hospital of Linhai from January 2016 to July 2017 were enrolled in the study.The colposcopy was used to get cervical biopsy lesions for pathological diagnosis, immunohistochemical staining was used to detect P16 and Ki-67 proteins.@*Results@#There were up to 66 patients with HPV-16 positive, 34 cases with HPV-18 positive, other 20 cases with 16 and 18 positive; 6 cases of atypical squamous epithelial cells(ASC), 46 cases of low-grade squamous intraepithelial lesions (LSIL), 60 cases of high-grade SIL(HSIL), other 8 cases of cervical squamous cell carcinoma(SCC) were diagnosed by TCT; 5 cases of inflammation, 105 cases of CIN and 10 cases of SCC by pathologically diagnosis.The positive rates of P16 and Ki-67 proteins were gradually increased in patients with inflammation, CIN and SCC[0(0/5), 36.2%(38/105), 70.0%(7/10), χ2=4.382, P=0.036; 0(0/5), 40.0%(42/105), 80.0%(8/10), χ2=5.945, P=0.015]. The patients with CIN were followed up for 21~36 months, median time 29.5 months.Twenty-six patients progressed to SCC, at the end of follow-up, the positive rates of P16 and Ki-67 proteins in patients with SCC were significantly higher than those in the CIN patients without progress[61.5%(16/26) vs.39.2%(31/79), χ2=3.934, P=0.047; 69.2%(18/26) vs.41.8%(33/79), χ2=5.905, P=0.015].@*Conclusion@#The positive expression rates of P16 and Ki-67 proteins in cervical biopsy tissues are significantly higher in patients with high-risk HPV infection and abnormal TCT, which in patients with pathologically diagnosis of SCC are higher than CIN patients, the same results in CIN patients with SCC progression than non-progressing patients, suggesting that P16 and Ki-67 proteins are of great value in predicting occurrence of early cervical cancer.

Clinical value of P16 and Ki-67 expressions in cervical biopsy for predicting early cervical cancer in patients with high-risk HPV infection / 中国基层医药

Objective To explore the positive expressions of P 16 and Ki -67 proteins in cervical biopsy lesions of patients with high-risk HPV infection and abnormal detection of liquid -based thin-layer cytology test (TCT),and the clinical value in predicting occurrence of early cervical cancer .Methods A total of 120 patients with high-risk HPV infection(16 and 18 subtypes positive) and abnormal TCT in the Traditional Chinese Medicine Hospital of Linhai from January 2016 to July 2017 were enrolled in the study.The colposcopy was used to get cervical biopsy lesions for pathological diagnosis ,immunohistochemical staining was used to detect P 16 and Ki-67 proteins. Results There were up to 66 patients with HPV-16 positive,34 cases with HPV-18 positive,other 20 cases with 16 and 18 positive;6 cases of atypical squamous epithelial cells (ASC),46 cases of low-grade squamous intraepithelial lesions (LSIL),60 cases of high-grade SIL(HSIL),other 8 cases of cervical squamous cell carcinoma (SCC) were diagnosed by TCT;5 cases of inflammation,105 cases of CIN and 10 cases of SCC by pathologically diagnosis.The positive rates of P16 and Ki-67 proteins were gradually increased in patients with inflammation ,CIN and SCC[0(0／5),36.2%(38／105),70.0%(7／10),χ2 ＝4.382,P＝0.036;0 (0／5),40.0%(42／105),80.0%(8／10),χ2 ＝5.945,P＝0.015].The patients with CIN were followed up for 21～36 months,median time 29.5 months.Twenty-six patients progressed to SCC ,at the end of follow-up,the positive rates of P16 and Ki-67 proteins in patients with SCC were significantly higher than those in the CIN patients without progress [61.5%(16／26) vs.39.2%(31／79), χ2 ＝3.934,P＝0.047;69.2%(18／26) vs.41.8%(33／79),χ2 ＝5.905,P＝0.015].Conclusion The positive expression rates of P16 and Ki-67 proteins in cervical biopsy tissues are significantly higher in patients with high -risk HPV infection and abnormal TCT ,which in patients with pathologically diagnosis of SCC are higher than CIN patients,the same results in CIN patients with SCC progression than non -progressing patients,suggesting that P16 and Ki-67 proteins are of great value in predicting occurrence of early cervical cancer .

Definitive radiochemotherapy or initial surgery for oropharyngeal cancer : To what extent can p16 expression be used in the decision process?

BACKGROUND: The decision between definitive radio(chemo)therapy (RCT) or a surgical strategy, i. e. surgery ± adjuvant radio(chemo)therapy for optimal treatment of oropharyngeal cancer is highly debated. Human papillomavirus(HPV)-related tumours are a distinct entity associated with p16 overexpression. While this represents a major prognostic factor, its predictive significance remains unknown. RESULTS: Among 183 consecutive unselected patients treated between 2009 and 2013 with a state-of-the-art surgical procedure ± adjuvant radio(chemo)therapy or definitive RCT including intensity-modulated radiotherapy, 3­year disease-free survival (DFS) was 74 vs. 57%, respectively (p = 0.007). When focusing on p16+ patients (49%), there was no significant difference in tumour control rate between surgery ± radio(chemo)therapy and the definitive RCT group (3-year DFS 83 vs. 82%, respectively; p = 0.48). However, delayed severe dysphagia was significantly lower in favour of definitive RCT: 35 vs. 4%, respectively; p = 0.0002. CONCLUSION: Our results highlight distinct outcomes after definitive RCT or initial surgical treatment according to p16 status, which should thus be considered during the decision process.

[Significance of HPV and cell cycle related proteins in basaloid squamous cell carcinoma of the larynx].

Objective: To study the significance of HPV and cell cycle related proteins in basaloid squamous cell carcinoma (BSCC) of the larynx. Methods: Twenty-nine cases of laryngeal BSCC from Beijing Tongren Hospital from January 2005 to December 2011 were reviewed. HPV typing by polymerase chain reaction-reverse dot blot (PCR-RDB) and p53, Ki-67, p16, p21 and cyclin D1 expression by immunohistochemistry were performed. The relationship between these indicators, various pathologic parameters (TNM, tumor size, tumor site and lymph node metastasis) and HPV status was analyzed. Results: There were 27 male and 2 female patients. The median age was 62 years. Lymph node metastasis and supraglottic tumor location were slightly higher than that of "usual" SCC, but not statistically significant (P>0.05). HPV DNA was detected in 27.6% (8/29) of the laryngeal BSCC, and all were HPV16. The expression of HPV was not related to age, alcohol consumption, tumor stage and tumor size. p53 was expressed in 31.0%(9/29) of laryngeal BSCC, and these cases were more likely supraglottic and had lymph node metastases (P<0.05). p16 staining was seen in 24.1% (7/29) of laryngeal BSCC, and these cases showed slightly higher rate of lymph node metastasis compared to p16 negative cases. The expression rates of p21 and cyclinD1 were 27.6% (8/29) and 69.0%(20/29), respectively, which were not related to age, tumor size, stage, lymph node metastasis, smoking and drinking. There were only 3 p16+ /HPV+ cases, which showed higher p21 and Ki-67 index compared to the HPV negative group (P<0.05). Conclusion: Some laryngeal BSCC expresses HPV DNA, possibly indicating an association with HPV; but p16 expression is not a reliable indicator for HPV infection.

Significance of HPV and cell cycle related proteins in basaloid squamous cell carcinoma of the larynx / 中华病理学杂志

Objective@#To study the significance of HPV and cell cycle related proteins in basaloid squamous cell carcinoma (BSCC) of the larynx.@*Methods@#Twenty-nine cases of laryngeal BSCC from Beijing Tongren Hospital from January 2005 to December 2011 were reviewed. HPV typing by polymerase chain reaction-reverse dot blot (PCR-RDB) and p53, Ki-67, p16, p21 and cyclin D1 expression by immunohistochemistry were performed. The relationship between these indicators, various pathologic parameters (TNM, tumor size, tumor site and lymph node metastasis) and HPV status was analyzed.@*Results@#There were 27 male and 2 female patients. The median age was 62 years. Lymph node metastasis and supraglottic tumor location were slightly higher than that of "usual" SCC, but not statistically significant (P>0.05). HPV DNA was detected in 27.6% (8/29) of the laryngeal BSCC, and all were HPV16. The expression of HPV was not related to age, alcohol consumption, tumor stage and tumor size. p53 was expressed in 31.0%(9/29) of laryngeal BSCC, and these cases were more likely supraglottic and had lymph node metastases (P<0.05). p16 staining was seen in 24.1% (7/29) of laryngeal BSCC, and these cases showed slightly higher rate of lymph node metastasis compared to p16 negative cases. The expression rates of p21 and cyclinD1 were 27.6% (8/29) and 69.0%(20/29), respectively, which were not related to age, tumor size, stage, lymph node metastasis, smoking and drinking. There were only 3 p16+ /HPV+ cases, which showed higher p21 and Ki-67 index compared to the HPV negative group (P<0.05).@*Conclusion@#Some laryngeal BSCC expresses HPV DNA, possibly indicating an association with HPV; but p16 expression is not a reliable indicator for HPV infection.

[Sorting role of p16(INK4a)/Ki-67 double immunostaining in the cervical cytology specimens of ASCUS and LSIL cases].

Objective: To investigate the sorting effect of p16(INK4a)/Ki-67 double immunostaining method in patients with atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesion (LSIL) cytology results. Methods: Four-hundred and twenty cases collected during April 2014 to February 2015 of cervical cytology of ASCUS (n=318) and LSIL (n=102) were selected, and residual liquid-based cytology specimens were used for p16(INK4a)/Ki-67 double immunostaining. The sensitivity and specificity of the detection of cervical precancerous lesions and cervical cancer were calculated, and the results were compared with high risk HPV. Taking histological follow-up as the gold standard, the test was considered positive when at least one cell exhibited p16(INK4a)/Ki-67 co-staining, without requirement of adjunct morphologic interpretation of positive cells. Results: Further screening CIN2+ in cytology ASCUS and LSIL group , the sensitivity of p16(INK4a)/Ki-67 double immunostaining was slightly lower than high risk HPV (84.2% vs. 94.7%), while the specificity was higher (84.0% vs. 53.9%). For ASCUS patients, the sensitivity of p16(INK4a)/Ki-67 double immunostaining and high risk HPV was 82.6% and 91.3%, and the specificity was 88.8% and 63.7%, respectively. For LSIL patients, the sensitivity of p16(INK4a)/Ki-67 double immunostaining and high risk HPV was 86.7% and 100.0%, and the specificity was 67.8% and 20.7%, respectively. For patients younger and older than 30 years, specificity of p16(INK4a)/Ki-67 double immunostaining was both higher than that of high risk HPV (80.8% vs. 42.3%; 84.6% vs. 56.9%). Conclusions: p16(INK4a)/Ki-67 double immunostaining can effectively identify the high risk population in ASCUS or LSIL, with higher specificity than high risk HPV test. p16(INK4a)/Ki-67 double immunostaining may benefit patients younger than 30 years of age as a preliminary or potential cytology-combining screening tool.

Sorting role of p16INK4a/Ki-67 double immunostaining in the cervical cytology specimens of ASCUS and LSIL cases / 中华病理学杂志

Objective@#To investigate the sorting effect of p16INK4a/Ki-67 double immunostaining method in patients with atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesion (LSIL) cytology results.@*Methods@#Four-hundred and twenty cases collected during April 2014 to February 2015 of cervical cytology of ASCUS (n=318) and LSIL (n=102) were selected, and residual liquid-based cytology specimens were used for p16INK4a/Ki-67 double immunostaining. The sensitivity and specificity of the detection of cervical precancerous lesions and cervical cancer were calculated, and the results were compared with high risk HPV. Taking histological follow-up as the gold standard, the test was considered positive when at least one cell exhibited p16INK4a/Ki-67 co-staining, without requirement of adjunct morphologic interpretation of positive cells.@*Results@#Further screening CIN2+ in cytology ASCUS and LSIL group , the sensitivity of p16INK4a/Ki-67 double immunostaining was slightly lower than high risk HPV (84.2% vs. 94.7%), while the specificity was higher (84.0% vs. 53.9%). For ASCUS patients, the sensitivity of p16INK4a/Ki-67 double immunostaining and high risk HPV was 82.6% and 91.3%, and the specificity was 88.8% and 63.7%, respectively. For LSIL patients, the sensitivity of p16INK4a/Ki-67 double immunostaining and high risk HPV was 86.7% and 100.0%, and the specificity was 67.8% and 20.7%, respectively. For patients younger and older than 30 years, specificity of p16INK4a/Ki-67 double immunostaining was both higher than that of high risk HPV (80.8% vs. 42.3%; 84.6% vs. 56.9%).@*Conclusions@#p16INK4a/Ki-67 double immunostaining can effectively identify the high risk population in ASCUS or LSIL, with higher specificity than high risk HPV test. p16INK4a/Ki-67 double immunostaining may benefit patients younger than 30 years of age as a preliminary or potential cytology-combining screening tool.

Expression of programmed death-ligand 1, topoisomerase Ⅱα and p16 proteins in cervical cancer and their significances / 肿瘤研究与临床

Objective To analyze the expression of programmed death ligand 1 (PD-L1),p16 proteins and DNA topoisomerase Ⅱ α (TOPO Ⅱ α) in cervical cancer and their clinical significances.Methods A total of 181 paraffin-embedded operating specimens and the biopsy including 106 cervical squamous cell carcinoma,30 high grade cervical intraepithelial neoplasia (CIN),30 low grade CIN and 33 normal cervical were selected from January 2014 to December 2015.An immunohistochemical EnVision method was used to detect the expression of PD-L1,p16 and TOPO Ⅱ α.Results The positive rate of PD-L1 expression in cervical squamous cell carcinoma was 66.0 ％ (70/106),but it was not expressed in high grade CIN,low grade CIN and normal cervical tissues,the differences were statistically significant (all P =0.000).Whereas,in squamous cell carcinoma,the positive rate of PD-L1 in poor differentiation group [77.3 ％ (51/66)] was higher than that in middle or high differentiation group [56.5 ％ (21/40)],and the difference was statistically significant (x2 =7.02,P =0.01).According to the FIGO stage,the positive rate in stage Ⅰ-Ⅱ group [54.9 ％ (28/51)] was lower than that in stage Ⅲ-Ⅳ group [76.4 ％ (42/55)],and the difference was statistically significant (x2 =5.47,P =0.02).But there was no statistical significance in lesion size or lymph node metastasis (both P ＞ 0.05).The positive rate of p16 in cervical squamous cell carcinoma was 99.1％ (105/ 106),in high grade CIN was 86.7 ％ (26/30),in low grade CIN was 26.7 ％ (8/30),and no expression in normal cervical tissues,the differences were statistically significant (all P ＜ 0.001).The expression of p16 had no relationship with histological differentiation,FIGO stage,lesion size,and lymph node metastasis (all P ＞ 0.05).The positive expression rate of TOPO Ⅱ α in cervical squamous cell carcinoma was 97.2 ％ (103/106),compared with that in high grade CIN [73.3 ％ (22/30)],low grade CIN [66.7 ％ (20/30)],and normal cervical tissues (no expression),the differences were statistically significant (all P =0.000).But there was nosignificant difference in the positive expression rate of TOPOⅡ α between high grade and low grade CIN (x2 =0.32,P =0.570).The expression of TOPO 1Ⅱ α had no relationship with histological differentiation,FIGO stage,lesion size and lymph node metastasis (all P ＞ 0.05).Conclusion The expression of PD-L1,p16 and TOPO Ⅱ α is associated with the invasion of cervical cancer.PD-L1 may be an immune checkpoint for the treatment of patients with invasive cervical cancer.

Expression of Ki-67 and P16 INK4a in chemically-induced perioral squamous cell carcinomas in mice / Expressão KI-67 e P16 INK4a em carcinomas espinocelulares periorais quimicamente induzidos em camundongos

ABSTRACT Objective: to evaluate the influence of Ki-67 and P16INK4a proteins immunohistochemical expressions on the clinical and morphological parameters of perioral squamous cell carcinoma induced with 9,10-dimethyl-1,2-benzanthracene (DMBA) in mice. Methods: we topically induced the lesions in the oral commissure of ten Swiss mice for 20 weeks, determining the time to tumors onset and the average tumor volume up to 26 weeks. In histopathological analysis, the variables studied were histological malignancy grade and the immunohistochemical expression of Ki-67 and P16INK4a proteins. The correlation between variables was determined by application of the Spearman correlation test. Results: the mean time to onset of perioral lesions was 21.1 ± 2.13 weeks; mean tumor volume was 555.91 ± 205.52 mm3. Of the induced tumors, 80% were classified as low score and 20% high score. There was diffuse positivity for Ki-67 in 100% of lesions - Proliferation Index (PI) of 50.1 ± 18.0. There was a strong direct correlation between Ki-67 immunoreactivity and tumor volume (R = 0.702) and a low correlation with the malignancy score (R = 0.486). The P16INK4a protein expression was heterogeneous, showing a weak correlation with tumor volume (R = 0.334). There was no correlation between the immunohistochemical expression of the two proteins studied. Conclusion: in an experimental model of DMBA-induced perioral carcinogenesis, tumor progression was associated with the tumor proliferative fraction (Ki-67 positive cells) and with tumor histological grading, but not with P16INK4a expression.

RESUMO Objetivo: avaliar a influência da expressão imuno-histoquímica das proteínas Ki-67 e p16INK4a sobre parâmetros clínico-morfológicos em carcinomas espinocelulares periorais quimicamente induzidos com 9,10-dimetil-1,2-benzantraceno (DMBA) em modelo murino. Métodos: as lesões foram induzidas topicamente na comissura labial de dez camundongos Swiss durante 20 semanas, sendo determinado o momento de surgimento dos tumores e volume tumoral médio até 26 semanas. Na análise histopatológica, as variáveis estudadas foram gradação histológica de malignidade tumoral e expressão imuno-histoquímica das proteínas Ki-67 e p16INK4a. A correlação entre as variáveis estudadas foi determinada pela aplicação do teste de correlação de Spearman. Resultados: o tempo médio de surgimento das lesões periorais foi 21,1±2,13 semanas. Volume tumoral médio foi de 555,91±205,52mm3. Dos tumores produzidos, 80% foram classificados como de baixo escore e 20%, alto escore. Evidenciou-se positividade difusa para Ki-67 em 100% das lesões - índice de marcação (PI) de 50,1±18,0. Verificou-se correlação direta forte entre a imunoexpressão do Ki-67 e o volume tumoral (R=0,702) e fraca correlação com o escore de malignidade (R=0,486). A expressão da proteína p16INK4a foi heterogênea, mostrando fraca correlação com o volume tumoral (R=0,334). Não houve correlação entre a expressão imuno-histoquímica das duas proteínas estudadas. Conclusão: Em modelo experimental de carcinogênese perioral DMBA-induzida, a progressão tumoral está associada à fração proliferativa do tumor (células ki-67 positivas) e com a gradação histológica tumoral, porém não com a expressão da p16INK4a.

Effects of HBV X protein on expression and promoter methylation of p16 tumor suppressor gene / 临床肝胆病杂志

ObjectiveTo explore the effects of hepatitis B virus X protein (HBx) on the expression and promoter methylation of the p16 tumor suppressor gene, and to investigate the epigenetic role of HBx in the development and progression of hepatitis B virus (HBV)-associated hepatocellular carcinomas (HCC). MethodsExperiments were performed in the human hepatoblastoma cell line HepG2, HepG2 cells expressing green fluorescent protein (HepG2/GFP), and HepG2 cells stably expressing GFP-HBx fusion protein (HepG2/GFP-HBx). Western blot was used to determine the expression levels of the p16 protein in HepG2 cells, HepG2/GFP cells, and HepG2/GFP-HBx cells. HepG2/GFP-HBx cells were treated with a universal inhibitor of DNA methyltransferase (DNMT), 5-aza-2'-deoxycytidine (5-aza-2'-dC). Methylation-specific polymerase chain reaction (MSP) was used to determine the promoter methylation of the p16 tumor suppressor gene in HepG2 cells, HepG2/GFP cells, and HepG2/GFP-HBx cells treated with or without 5-aza-2′-dC. Multiple-group comparison was made by analysis of variance. ResultsAccording to the results of Western blot, HepG2/GFP-HBx cells had a significantly lower expression level of the p16 protein than HepG2 cells and HepG2/GFP cells (P=0.0007; P=00014); there was no significant difference in the expression level of the p16 protein between HepG2/GFP and HepG2 cells (P＞0.05). The MSP assay revealed partial CpG methylation in the p16 promoter region in HepG2/GFP-HBx cells. No promoter methylation was detected in HepG2 cells or HepG2/GFP cells. Non-methylation in the p16 promoter region was restored in HepG2/GFP-HBx cells treated with 5-aza-2′-dC. ConclusionIn the hepatoblastoma cell line, HBx down-regulates the expression of the p16 tumor suppressor gene by inducing methylation in its promoter region. The DNMT inhibitor, 5-aza-2′-dC, restores non-methylation in the p16 promoter region. The reversible modification provides new insights for the treatment and prevention of HBV-associated HCC.

[HPV infection in oral, pharyngeal and laryngeal papillomas]. / HPV-Infektion in oralen, pharyngealen und laryngealen Papillomen.

BACKGROUND: HPV infections play a major role in the pathogenesis of squamous cell carcinomas of the head and neck. Regarding benign papillomas, the role of HPV is still uncertain. MATERIALS AND METHODS: To clarify this issue, 100 exophytic papillomas of the oral cavity, pharynx and larynx were subjected to histopathological and molecular pathological examination. Excision biopsies were taken from 62 male and 38 female patients with an age range of 18 to 87 years. Biopsies were tested for p16 expression by immunohistochemistry and analyzed for HPV subtypes 6/11 (low-risk), 16/18 and 31/33/53 (high-risk) by chromogenic in situ hybridization. RESULTS: HPV infections were verified molecularly in 34 % of biopsies; in all cases with the low-risk HPV subtypes 6/11. Only one case showed infection with both 6/11 and 31/33/53 subtypes, but not subtype 16/18; whereas expression of p16 was found in 67 %. The rate of positive molecular verification of HPV infection (in situ hybridization) was highest in the laryngeal lesions with 61.1 %, followed by the oral cavity with 52.9 %, and lowest in pharyngeal lesions (21.5 %). Recurrent papillomas were seen in 18 cases (18 %), of which 14 were molecularly positive for HPV (in situ hybridization). A correlation between inflammatory infiltration and HPV infection could be verified in 82 %. CONCLUSION: Our data demonstrate an important role of HPV infection for the development of benign papillomas of the head and neck region. Furthermore, there is a positive correlation between HPV infection and recurrent papillomas. Therefore, a molecular morphological HPV analysis of papillomas could provide important prognostic data.

Biologia molecular no carcinoma de endométrio: uma revisão / Molecular biology in endometrial carcinoma: a review

Este artigo objetivou oferecer uma visão atual do papel dos marcadores gênicos no carcinoma de endométrio. Os principais genes descritos são o TP53, o Bcl-2, o c-erbB2 e o p16. Nos últimos anos, com a ampliação do conhecimento na área de biologia molecular, tem sido sugerido que os marcadores biológicos possam ser tão ou mais importantes do que os fatores prognósticos convencionais.

The main of this study is offer the present situation of genic markers in endometrial carcinoma. The principal genes have been described are TP53, Bcl-2, c-erbB2 and p16. In the last few years, thanks to improvements in molecular biology, some biological markers have been suggested to be as important as or more important than conventional prognostic factors.

Expression and significance of p15, p16, p21 in Uygur patients with psoriasis in Xinjiang / 中国医师杂志

Objective To investigate the expressions of p15,p16,and p21 in skin of uygur patients with psoriasis and its significance in the development of psoriasis.Methods The expressions of p15,p16,and p21 were studied with immunohistochemical method in the Xinjiang Uygur psoriatic and normal Uygur skins.Results The positive expression rate of p16 gene was 12.5％ in skin lesions of patients with psoriasis Uygur,and 66.67％ in the normal group.The positive expressions of p15,and p21 genes in skin lesions of patients with psoriasis and healthy Uygur population were higher.Conclusions There is a significant correlation between the development of uygur psoriasis vulgaris and abnormal expression of p16.

Methylation status of p16 gene and expressions of related genes in keloid tissue and cultured keloid fibroblasts / 中华皮肤科杂志

Objective To explore the role of p16 gene methylation in fibroblasts in the occurrence and development of keloid.Methods Skin tissue specimens were resected from the lesions of patients with keloid and normal skin of healthy human controls.Fibroblasts were isolated from these tissue specimens and subjected a primary culture.An immunohistochemical analysis was performed to measure the expression of p16 protein in tissue specimens,real-time fluorescence-based quantitative PCR to determine the mRNA expression level (expressed as 2-△△Ct) of p 16 and DNA methyltransferases (DNMTs) in fibmblasts,and bisulfite sequencing PCR (BSP) to estimate the methylation status of p16 gene in the tissue specimens and primary fibroblasts.Results The keloid fibroblasts (KFbs) showed significandy lower mRNA expression of p16 gene (0.64 ± 0.18 vs.1.92 ± 0.23,t =10.54,P＜ 0.05),but significantly higher mRNA expressions of 3 DNMTs (DNMT1:2.58 ± 0.23 vs.1.13 ± 0.21,t =11.22,P ＜ 0.05; DNMT3A:4.87 ± 0.46 vs.2.38 ± 0.32,t =10.81,P＜ 0.05; DNMT3B:1.57 ± 0.12 vs.0.57 ± 0.16,t =12.45,P＜ 0.05) compared with the normal fibmblasts (NFbs).The DNA methylation rate in the p16 gene promoter region was significantly increased in keloid tissue (1.81％ ± 0.46％) and KFbs (3.15％ ± 0.94％) compared with normal skin tissue (0.90％ ± 0.35％,F =14.23,P＜ 0.01) and NFbs (0.17％ ± 0.29％,F=37.62,P＜ 0.01).Conclusions The methylation and low expression of p16 gene in KFbs may be associated with the uncontrolled growth of keloid,and DNMTs may play a role in the pathogenesis of keloid.