Evaluation of p16/Ki-67 dual-stained cytology as triage test for high-risk human papillomavirus-positive women

Abstract

The aim of this study was to evaluate the clinical utility of p16/Ki-67 dual staining, for the identification of CIN in high-risk HPV-positive women from a non-responder screening cohort. P16/Ki-67 dual staining, Pap cytology, and HPV16/18 genotyping were performed on physician-taken liquid-based samples from 495 women who tested high-risk HPV positive on self-sampled material (PROHTECT-3B study). Different triage strategies involving p16/Ki-67 dual staining were evaluated for sensitivity, specificity, and predictive value for ≥CIN2 and ≥CIN3, and compared to Pap cytology with a threshold of atypical cells of undetermined significance. Centrally revised histology or an adjusted endpoint with combined high-risk HPV negative and cytology negative follow-up at 6 months was used as gold standard. Pap cytology (threshold atypical cells of undetermined significance) triage of high-risk HPV-positive samples showed a sensitivity of 93% (95% confidence interval: 85-98) with a specificity of 49% (95% confidence interval: 41-56) for ≥CIN3. Three triage strategies with p16/Ki-67 showed a significantly increased specificity with similar sensitivity. P16/Ki-67 triage of all high-risk HPV-positive samples had a sensitivity of 92% (95% confidence interval: 84-97) and a specificity of 61% (95% confidence interval: 54-69) for ≥CIN3. Applying p16/Ki-67 triage to only high-risk HPV-positive women with low-grade Pap cytology showed a similar sensitivity of 92% (95% confidence interval: 84-97), with a specificity for ≥CIN3 of 64% (95% confidence interval: 56-71). For high-risk HPV-positive women with low-grade and normal Pap cytology, triage with p16/Ki-67 showed a sensitivity of 96% (95% confidence interval: 89-99), and a specificity of 58% (95% confidence interval: 50-65). HPV16/18 genotyping combined with Pap cytology showed a sensitivity and specificity for ≥CIN3 similar to Pap cytology with an atypical cells of undetermined significance threshold. Because the quality of Pap cytology worldwide varies, and differences in sensitivity and specificity are limited between the three selected strategies, p16/Ki-67 triage of all high-risk HPV-positive samples would be the most reliable strategy in triage of high-risk HPV-positive women with an increased specificity and similar sensitivity compared with Pap cytology triage.

The role of p16 as putative biomarker for cervical neoplasia: A controversial issue?

“…14 Several studies have shown increased sensitivity and specificity for cervical cancer screening using p16 staining in conjunction with other biomarkers, such as p16/Ki-67 15,16,[24][25][26] and Survivin, the inhibitor of apoptosis. 27 …”

Approaches to triage optimization in HPV primary screening: Extended genotyping and p16/Ki‐67 dual‐stained cytology—Retrospective insights from ATHENA

“…21 In ATHENA, and other studies, DS testing demonstrated better performance (improved sensitivity and at least equal specificity) compared to cytology for the cross-sectional detection of ≥CIN3 in hrHPV-positive women. 20,[22][23][24][25][26][27][28][29][30][31] However, like Pap cytology, DS testing is not integrated into the first-line HPV screening test, and both Pap cytology and DS testing require the preparation of a slide for subjective microscopic assessment by highly trained professionals.…”

Association of p16 (CDKN2A) polymorphisms with the development of HPV16‐related precancerous lesions and cervical cancer in the Greek population

“…The E7 oncoprotein disrupts the pRb/E2F suppressor complex, thus leading to the release of the E2F transcriptional factor which subsequently stimulates the transition of cell cycle from G1 to S phase . Furthermore, the E7‐mediated degradation of pRb triggers the overexpression of the nuclear protein p16 . In particular, inactivation of pRb inhibits the negative feedback control of p16 and subsequently leads to the excessive expression of p16 in order to eliminate the prolonged proliferation of HPV infected cells …”

“…Furthermore, the E7‐mediated degradation of pRb triggers the overexpression of the nuclear protein p16 . In particular, inactivation of pRb inhibits the negative feedback control of p16 and subsequently leads to the excessive expression of p16 in order to eliminate the prolonged proliferation of HPV infected cells …”