Top

Diagnostic Pathology

Published in:

Open Access 01-12-2021 | Research

Accuracy of HPV E6/E7 mRNA examination using in situ hybridization in diagnosing cervical intraepithelial lesions

Authors: Chang Hui, Huimin Bai, Jun Liu, Xuerong Lu, Shuzhen Wang, Zhenyu Zhang, Mulan Jin, Yue Wang, Yuzhen Liu

Published in: Diagnostic Pathology | Issue 1/2021

Abstract

Background

The consistency of pathologists in the diagnosis of cervical intraepithelial neoplasia (CINs) is not ideal, especially between low- and high-grade squamous intraepithelial lesions (LSIL and HSIL). This study was aimed to explore efficient strategies for the grading of CINs.

Methods

The medical records of patients with high risk human papillomavirus (HR-HPV) infections who had underwent cervical biopsy or conization from April 2018 to April 2019 in Beijing Chao-Yang Hospital were collected and examined. The HR-HPV E6/E7 mRNA in the tissues of patients with CINs was detected using RNAscope chromogenic in situ hybridization (RISH). Immunohistochemistry (IHC) was performed to evaluate the expression of p16INK4a (P16) and Ki67.

Results

HR-HPV E6/E7 mRNA signals were detected in 3/27 (11.1 %) of CIN 1, and in 32/33 (97.0 %) of CIN 2/3. Most of the staining patterns (27/32, 84.4 %) had a full-thickness epithelial layer staining with weak-to-strong nuclear and cytoplasmic dot-like signals in CIN 2/3, and there were also few special staining patterns that were significantly different from the others. A number of indicators were compared between LSIL and HSIL. There were statistically significant differences in E6/E7 mRNA, p16, Ki67 and cytology between the two groups (P < 0.05). According to the logistic regression analysis, merely E6/E7 mRNA positivity was significantly associated with CIN2/3 (OR: 52.53, 95 % CI, P < 0.05). In the detection of CIN 2/3, the sensitivity and specificity of HPV E6/E7 mRNA alone was not significantly inferior to that of its different combinations with Ki67, p16 and cytology (all, P > 0.05).

Conclusions

RISH is efficient in grading of CINs. The HPV E6/E7 mRNA expression might reflect the phase HPV infections, and its positive pattern might predict the development direction of CINs, providing the possibility to realize more accurate treatments for patients.

Arbyn M, Weiderpass E, Bruni L, de Sanjosé S, Saraiya M, Ferlay J, et al. Estimates of incidence and mortality of cervical cancer in 2018: a worldwide analysis. Lancet Glob Health. 2020;8(2):e191–e203.

Bosch FX, Lorincz A, Muñoz N, Meijer C, Shah KV. The causal relation between human papillomavirus and cervical cancer. J Clin Pathol. 2002;55:244–65.CrossRefPubMed

Darragh TM, Colgan TJ, Thomas Cox J, Heller DS, Henry MR, Luff RD, et al. The Lower Anogenital Squamous Terminology Standardization project for HPV-associated lesions: background and consensus recommendations from the College of American Pathologists and the American Society for Colposcopy and Cervical Pathology. Int J Gynecol Pathol. 2013;32:76–115.CrossRef

de Sanjose S, Brotons M, Pavon MA. The natural history of human papillomavirus infection. Best Pract Res Clin Obstet Gynaecol. 2018;47:2–13.CrossRefPubMed

Carreon JD, Sherman ME, Guillen D, Solomon D, Herrero R, Jeronimo J, et al. CIN2 is a much less reproducible and less valid diagnosis than CIN3: results from a histological review of population-based cervical samples. Int J Gynecol Pathol. 2007;26:441–6.CrossRef

Dalla Palma P, Giorgi Rossi P, Collina G, Buccoliero AM, Ghiringhello B, Gilioli E, et al. The reproducibility of CIN diagnoses among different pathologists: data from histology reviews from a multicenter randomized study. Am J Clin Pathol. 2009;132:125–32.CrossRef

Carozzi F, Confortini M, Dalla Palma P, Del Mistro A, Gillio-Tos A, De Marco L, et al. New Technologies for Cervival Cancer Screening (NTCC) working group use of p16-INK4A overexpression to increase the specificity of human papillomavirus testing: a nested substudy of the NTCC randomised controlled trial. Lancet Oncol. 2008;9:937–45.CrossRef

Murphy N, Ring M, Killalea A, Uhlmann V, O’Donovan M, Mulcahy F, et al. p16INK4A as a marker for cervical dyskaryosis: CIN and cGIN in cervical biopsies and ThinPrep™ smears. J Clin Pathol. 2003;56:56–63.CrossRefPubMed

Horn L-C, Reichert A, Oster A, Arndal SF, Trunk MJ, Ridder R, et al. Immunostaining for p16INK4a used as a conjunctive tool improves interobserver agreement of the histologic diagnosis of cervical intraepithelial neoplasia. Am J Surg Pathol. 2008;32:502–12.CrossRef

10.

Guedes A, Brenna S, Coelho S, Martinez EZ, Syrjänen K, Zeferino L. p16INK4a expression does not predict the outcome of cervical intraepithelial neoplasia grade 2. Int J Gynecol Cancer. 2007;17:1099–103.CrossRefPubMed

11.

Sagasta A, Castillo P, Saco A, Torné A, Esteve R, Marimon L, et al. p16 staining has limited value in predicting the outcome of histological low-grade squamous intraepithelial lesions of the cervix. Mod Pathol. 2016;29:51.CrossRefPubMed

12.

Campbell LM, Pitta DR, De Assis AM, Derchain SFM, Campos EA, Sarian LOZ. Retrieval of HPV oncogenes E6 and E7 mRNA from cervical specimens using a manual open technology protocol. Springerplus. 2013;2:473.CrossRefPubMed

13.

Dockter J, Schroder A, Eaton B, Wang A, Sikhamsay N, Morales L, et al. Analytical characterization of the APTIMA® HPV assay. J Clin Virol. 2009;45:39–47.CrossRef

14.

Cassidy A, Jones J. Developments in in situ hybridisation. Methods. 2014;70:39–45.CrossRefPubMed

15.

Anderson CM, Zhang B, Miller M, Butko E, Wu X, Laver T, et al. Fully automated RNAscope in situ hybridization assays for formalin-fixed paraffin-embedded cells and tissues. J Cell Biochem. 2016;117:2201–8.CrossRefPubMed

16.

Ukpo OC, Flanagan JJ, Ma X-J, Luo Y, Thorstad WL, Lewis JS. High-risk human papillomavirus E6/E7 mRNA detection by a novel in situ hybridization assay strongly correlates with p16 expression and patient outcomes in oropharyngeal squamous cell carcinoma. Am J Surg Pathol. 2011;35:1343–50.CrossRefPubMed

17.

Mirghani H, Casiraghi O, Guerlain J, Amen F, He M-X, Ma X-J, et al. Diagnosis of HPV driven oropharyngeal cancers: comparing p16 based algorithms with the RNAscope HPV-test. Oral Oncol. 2016;62:101–8.CrossRefPubMed

18.

Nayar R, Wilbur DC. The Pap test and Bethesda 2014. Cancer Cytopathol. 2015;123:271–81.CrossRefPubMed

19.

Massad LS, Einstein MH, Huh WK, et al. 2012 updated consensus guidelines for the management of abnormal cervical cancer screening tests and cancer precursors. J Lower Genit Tract Dis. 2013;17:S1–27.CrossRef

20.

Graham SV. The human papillomavirus replication cycle, and its links to cancer progression: a comprehensive review. Clin Sci (Lond). 2017;131:2201–21.CrossRef

21.

Wuerthner BA, Avila-Wallace M. Cervical cancer: screening, management, and prevention. Nurse Pract. 2016;41:18–23.CrossRef

22.

Kanthiya K, Khunnarong J, Tangjitgamol S, Puripat N, Tanvanich S. Expression of the p16 and Ki67 in cervical squamous intraepithelial lesions and cancer. Asian Pac J Cancer Prev. 2016;17:3201–6.

23.

Wentzensen N, Schwartz L, Zuna RE, Smith KM, Mathews C, Gold MA, et al. Performance of p16/Ki-67 immunostaining to detect cervical cancer precursors in a colposcopy referral population. Clin Cancer Res. 2012;18:4154–62.CrossRefPubMed

24.

Martin C, Oleary JJ. Histology of cervical intraepithelial neoplasia and the role of biomarkers. Best Pract Res Clin Obstet Gynaecol. 2011;25:605–15.CrossRef

25.

Zafereo M, Xu L, Dahlstrom KR, Viamonte CA, Elnaggar AK, Wei Q, et al. Squamous cell carcinoma of the oral cavity often overexpresses p16 but is rarely driven by human papillomavirus. Oral Oncol. 2016;56:47–53.CrossRefPubMed

26.

Castle PE, Adcock R, Cuzick J, Wentzensen N, Torrezmartinez NE, Torres SM, et al. Relationships of p16 Immunohistochemistry and other biomarkers with diagnoses of cervical abnormalities: implications for LAST terminology. Arch Pathol Lab Med. 2020;144(6):725–34.

27.

Liu YZ, Lu XP, Pan ZX, Zhang W, Chen ZR, Wang H, et al. Establishment of a novel method for primary culture of normal human cervical keratinocytes. Chin Med J. 2013;126:3344.

28.

Galgano MT, Castle PE, Atkins KA, Brix WK, Nassau SR, Stoler MH. Using biomarkers as objective standards in the diagnosis of cervical biopsies. Am J Surg Pathol. 2010;34:1077–87.CrossRefPubMed

29.

Scholzen T, Gerdes J. The Ki-67 protein: from the known and the unknown. J Cell Physiol. 2000;182:311–22.CrossRef

30.

Kruse A, Baak JPA, De Bruin PC, Jiwa M, Snijders W, Boodt PJ, et al. Ki-67 immunoquantitation in cervical intraepithelial neoplasia (CIN): A sensitive marker for grading. J Pathol. 2001;193:48–54.CrossRefPubMed

31.

Ikenberg H, Bergeron C, Schmidt D, Griesser H, Alameda F, Angeloni C, et al. Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology: results of the PALMS study. J Natl Cancer Inst. 2013;105:1550–7.CrossRefPubMed

32.

Ozaki S, Zen Y, Inoue M. Biomarker expression in cervical intraepithelial neoplasia: potential progression predictive factors for low-grade lesions. Hum Pathol. 2011;42:1007–12.CrossRefPubMed

33.

Sobecki M, Mrouj K, Colinge J, Gerbe F, Jay P, Krasinska L, et al. Cell-cycle regulation accounts for variability in Ki-67 expression levels. Cancer Res. 2017;77:2722–34.CrossRefPubMed

34.

Doorbar J. Molecular biology of human papillomavirus infection and cervical cancer. Clin Sci. 2006;110:525–41.CrossRef

35.

Graham SV. The human papillomavirus replication cycle, and its links to cancer progression: a comprehensive review. Clin Sci. 2017;131:2201–21.CrossRef

36.

Evans MF, Peng Z, Clark KM, Adamson CS, Ma X, Wu X, et al. HPV E6/E7 RNA in situ hybridization signal patterns as biomarkers of three-tier cervical intraepithelial neoplasia grade. PLoS One. 2014;9(3):e91142.

37.

Moscicki A, Ma Y, Wibbelsman C, Darragh TM, Powers A, Farhat S, et al. Rate of and risks for regression of cervical intraepithelial neoplasia 2 in adolescents and young women. Obstet Gynecol. 2010;116:1373–80.CrossRefPubMed

38.

Zeng SY, Liang MR, Li LY, Wu YY. Comparison of the efficacy and complications of different surgical methods for cervical intraepithelial neoplasia. Eur J Gynaecol Oncol. 2012;33:257.PubMed

39.

Castle PE, Stoler MH, Solomon D, Schiffman M. The relationship of community biopsy-diagnosed cervical intraepithelial neoplasia grade 2 to the quality control pathology-reviewed diagnoses: an ALTS report. Am J Clin Pathol. 2007;127:805–15.CrossRefPubMed

Title: Accuracy of HPV E6/E7 mRNA examination using in situ hybridization in diagnosing cervical intraepithelial lesions
Authors: Chang Hui
Huimin Bai
Jun Liu
Xuerong Lu
Shuzhen Wang
Zhenyu Zhang
Mulan Jin
Yue Wang
Yuzhen Liu
Publication date: 01-12-2021
Publisher: BioMed Central
Published in: Diagnostic Pathology / Issue 1/2021
Electronic ISSN: 1746-1596
DOI: https://doi.org/10.1186/s13000-021-01072-9

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Accuracy of HPV E6/E7 mRNA examination using in situ hybridization in diagnosing cervical intraepithelial lesions

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2021

The clinicopathological relevance of uniform CD56 expression in anaplastic large cell lymphoma: a retrospective analysis of 18 cases

Placental pathology of the third trimester pregnant women from COVID-19

T cell exhaustion is associated with the risk of papillary thyroid carcinoma and can be a predictive and sensitive biomarker for diagnosis

Identification of factors related to immunotherapy efficacy and prognosis in patients with advanced head and neck squamous cell carcinoma

The many faces of solitary fibrous tumor; diversity of histological features, differential diagnosis and role of molecular studies and surrogate markers in avoiding misdiagnosis and predicting the behavior

Prognostic significance of the aberrant expression of neuroendocrine markers in melanomas