Characterization of p16 and E6 HPV-related proteins in uterine cervix high-grade lesions of patients treated by conization with large loop excision.

Cervical cancer and its precursor lesions represent a significant public health problem for developing and less-developed countries. Cervical carcinogenesis is strongly correlated with persistent high-risk human papillomavirus (HPV) infection, which is mostly associated with expression of the p16 and E6 HPV-related proteins. The aim of this present study was to determine the expression of the p16 and E6 proteins in females with high-grade lesions treated with conization, and to discuss the role of these proteins as prognostic markers following treatment. In total, 114 females were treated for high-grade cervical intraepithelial neoplasia (CIN, grades 2/3) by conization with large loop excision of the transformation zone (LLETZ). Following surgery, the patients returned within 30-45 days for post-operative evaluation. A follow-up was conducted every 6 months for 2 years. At each follow-up appointment, a Pap smear, colposcopy and HPV DNA test were performed. E6 and p16 immunohistochemical tests were conducted on the surgical specimens. The positive expression of p16 was correlated with the presence of lesions with increased severity in the surgical specimens (P= 0.0001). The expression of E6 did not demonstrate the same correlation (P=0.131). The HPV DNA hybrid, collected in the first post-operative consultation as a predictor of the cytological abnormalities identified at the 24-month follow-up assessment, presented a sensitivity of 55.6%, a specificity of 84.8%, a positive predictive value of 33.3% and a negative predictive value of 93.3%. The role of p16INK4A as a marker of CIN was also demonstrated; the expression of p16 and E6, however, did not appear to be of any prognostic value in predicting the clearance of high-risk HPV following conization. A negative hybrid capture test was correlated with a disease-free outcome.

Valor da captura híbrida para o papilomavírus humano (HPV) no seguimento de pacientes submetidos à conização do colo uterino devido a lesão intraepitelial de alto grau por cirurgia de alta frequência (CAF) / The value of the HPV DNA test on the follow-up of the patients treated for high grade cervical intraepithelial lesions by conization with large loop excision of the transformation zone (LLETZ)

INTRODUÇÃO: A lesão intraepitelial cervical de alto grau causada pelo HPV, precursora do câncer cervical, é facilmente diagnosticada e seu tratamento pode ser realizado de maneira ambulatorial, sem muitas complicações. Mesmo assim, o seguimento das pacientes tratadas deve ser feito de maneira criteriosa e sistemática para que a recorrência ou persistência da doença não passe despercebida. Conseguir identificar o grupo de pacientes com maior probabilidade de recorrência ou persistência da doença facilitaria sobremaneira esse seguimento, diminuindo o ônus econômico e psicológico que o seguimento generalizado produz. O objetivo deste trabalho é identificar marcadores que possam indicar o grupo de pacientes com maior possibilidade de recorrência da lesão intraepitelial cervical de alto grau. MÉTODOS: Neste estudo 114 mulheres com diagnóstico de lesão intraepitelial cervical de alto grau foram submetidas à conização cervical por cirurgia de alta frequência (CAF) no Setor de Patologia do Trato Genital Inferior (PTGI) da Divisão da Clínica Ginecológica do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP) no período entre março de 2006 e maio de 2009. O seguimento foi realizado a cada seis meses durante o período de 24 meses. No seguimento foi coletada a citologia cervical, captura híbrida para HPV e realizada a colposcopia. Foram avaliados os testes de captura híbrida para HPV coletadas durante o seguimento, imuno-histoquímicos para oncoproteína viral E6 e proteína p16 na peça cirúrgica como possíveis marcadores de recorrência. RESULTADOS: A avaliação anatomopatológica da peça cirúrgica diagnosticou 85 (74,6 por cento) casos de lesão intraepitelial cervical de alto grau e 29 (25,4 por cento) casos de lesão intraepitelial de baixo grau. Nessas peças, 45 (39,5 por cento) apresentaram expressão positiva para oncoproteína E6 e 69 (60,5 por cento) apresentaram expressão negativa para E6; 74 (64,9 por cento) apresentaram expressão positiva para p16 e 40 (35,1 por cento) apresentaram...

INTRODUCTION: The high grade cervical intraepithelial lesion caused by HPV, a pre-malignant condition, is easily diagnosed and its treatment can be done in outpatients without many complications. Nevertheless the patients follow-up must be done in a very systematic way to avoid any recurrence or persistence of the disease. To be able to identify the group of patients with higher rate of recurrence or persistence of the disease would make this follow-up much easier and decrease the economic and psychological burden of stressed outcome. The goal of our study is to identify markers that could indicate the group of patients more likely to recur. METHODS: In this study, 114 women diagnosed with high grade cervical intraepithelial lesion were treated with LLETZ at the Discipline of Gynecology, Faculty of Medicine, São Paulo University from March 2006 and May 2009. The follow-up visits after the treatment included Pap smear, HPV DNA test and colposcopy and occurred every 6 months for 24 months. The markers evaluated were the HPV DNA test collected during the follow-up and immunohistochemical tests performed on the surgical specimen: E6 oncoprotein and protein p16. RESULTS: We found 85 cases (74,6 percent) of HSIL and 29 cases (25,4 percent) of LSIL in the surgical specimen obtained with the LLETZ. The E6 oncoprotein was expressed in 45 (39,5 percent) and not expressed in 69 (60,5 percent) of the specimen; 74 (64,9 percent) expressed p16 and 40 (35,1 percent) didnt express p16. The E6 oncoprotein was not associated with severe presentation of the disease. The protein p16 was positive in 68 (80 percent) cases of diagnosed HSIL and negative in 23 (79,3 percent) cases with diagnosed LSIL or chronic cervicitis. The HPV DNA test collected at the first follow-up consult at 6 months presented a sensitivity of 83,3 percent specificity of 87,8 percent, positive predictive value (PPV) of 50 percent and negative predictive value (NPV) of 97,3 percent. Comparing the HPV DNA test collected at the first follow-up visit and the cervical cytology collected at the...

Evaluation of the combination of cytology and hybrid capture to safely predict the high-grade lesion status of patients treated with conization with large loop excision of the transformation zone.

OBJECTIVES: This study aimed to verify whether human papillomavirus (HPV) testing after conization treatment has some potential usefulness for predicting patients' outcome. STUDY DESIGN: One hundred and twenty women were treated for HSIL by conization with large loop excision of the transformation zone (LLETZ). Cytology, colposcopy-guided biopsy, and hybrid capture 2 (HC2) HPV DNA tests were performed before the surgical procedure and every 6 months for 2 years at follow-up. RESULTS: More than 90% of the patients tested positive for high-risk HPV prior to the surgical intervention. Six months after the cervical conization, 74.75% of the patients tested negative for high-risk HPV DNA, and 19.41% were positive. Of the women who were HC2 negative, 72 showed normal cytological smears, 3 ASC-US, 2 LSIL, and 1 HSIL. Of those who were HC2 positive, 8 showed normal smears, 2 ASC-US, 2 ASC-H, 5 LSIL, and 1 case had HSIL, AGC, and squamous cells invasive carcinoma. Clinically, the HSIL case with a negative HPV test did not show any sign of high-grade lesions, and the clinical follow-up did not show residual lesions. CONCLUSIONS: Negative HPV tests correlated with freedom from high-grade disease after 2 years of postconization follow-up, which strongly suggests that negative HPV tests predict the absence of cervical disease.