High Risk HPV E6/E7 Oncoprotein Expression in Women with High Grade Squamous Intraepithelial Lesion

 

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Abstract

Purpose To correlate the expression of high-risk HPV E6 mRNA with pap smear, colposcopy, and biopsy results in women with high grade squamous intraepithelial lesion (HSIL).

Methods A cross-sectional study was performed on women referred for primary care services after cytological diagnosis of HSIL. We evaluated the expression of E6/E7 mRNA of HPV types 16,18,31,33, and 45 and correlated the results with those of Pap smear, colposcopy, and biopsy. For amplification/detection of mRNA E6 / E7 we used NucliSENSEasyQ kit to detect HPV mRNA by polymerase chain reaction with primers/probes for HPV types 16, 18, 31, 33, and 45.

Results Out of 128 valid tests, the results of 30 (23.4%) tests were negative and 98 (70%) tests were positive. Only one type of HPV was detected in 87.7% of the E6/E7 mRNA positive cases. HPV16 was detected in 61.2% of the cases, followed by HPV33 (26.5%), HPV31 (17.3%), HPV18 (10%), and HPV45 (4.08%). Pap smear tests revealed that the E6/E7 test was positive in 107 (83.8%) women with atypical squamous cells – high grade (ASC-H), HSIL, or higher. The E6/E7 test was positive in 69 (57.5%) specimens presenting negative cytology results. When analyzing the association with colposcopy results, the frequency of positive E6/E7 results increased with the severity of the injury, ranging from 57.1% in women without colposcopy-detected injury to 86.5% in those with higher levels of colposcopy findings. Of the 111 women who underwent biopsy and E6/E7 testing, the E6/E7 test was positive in 84.7% of the women who presented with lesions of cervical intraepithelial neoplasia (CIN) grade 2 or higher. Finally, 41.2% of women with a negative biopsy presented a positive E6/E7 test.

Conclusions E6/E7 mRNA expression was higher in women with HSIL and CIN grade 2 or higher.

Resumo

Objetivo Correlacionar a expressão mRNAE6/E7 do HPV de alto risco com os exames de Papanicolau, colposcopia e biópsia em mulheres com lesão intraepitelial escamosa de alto grau (HSIL).

Métodos Estudo transversal com mulheres encaminhadas aos serviços de atenção primária com diagnóstico citológico de HSIL. Foi avaliada a expressão do mRNAE6/E7 dos tipos de HPV 16,18,31,33 e 45, correlacionando-se a expressão com os exames de Papanicolau, colposcopia e biópsia. Para a amplificação/detecção de mRNA de E6/E7 foi usado o kit NucliSENS EasyQ® HPV que detecta mRNA do HPV por meio da reação em cadeia da polimerase com primers/probes HPV dos tipos 16, 18, 31, 33 e 45.

Resultados Foram obtidos 128 testes válidos. Destes: 30 (23,4%) foram negativos e 98 (70%) dos testes foram positivos. Foi encontrado apenas um tipo de HPV em 87,7% dos positivos. O HPV16 foi o mais encontrado em 61,2%, seguido pelos HPV33 (26,5%); HPV31 (17,34%); HPV 18 (10,0%) e HPV (45 4,0%). Quanto ao exame de Papanicolau, o teste E6/E7 foi positivo em 107 (83,8%) das mulheres com ASC-H, HSIL ou superior, enquanto em citologia negativa foi encontrado um resultado positivo em 69 (57,5%) colposcopia. A frequência de teste E6/E7 positivo aumentou com a gravidade da lesão, detectada na colposcopia variando de 57,1% em mulheres sem lesão identificada em colposcopia até 86,5% naqueles com achado de colposcopia de grau maior. Das 111 mulheres que se submeteram a biópsia e o teste E6/E7, o teste foi positivo em 84,7% das que apresentaram lesão igual ou superior a NIC 2 (neoplasia intraepitelial cervical) e 41,2% daqueles com biópsia negativa.

Conclusões A expressão de E6, E7 RNAm ocorreu com maior frequência em lesões de alto grau citológica e em casos com biópsias de NIC2 ou maior.

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