Use of cyclin D1 in conjunction with human papillomavirus status to predict outcome in oropharyngeal cancer

Abstract

There is increasing use of multiple molecular markers to predict prognosis in human cancer. Our aim was to examine the prognostic significance of cyclin D1 and retinoblastoma (pRb) expression in association with human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma. Clinical records and specimens of 226 patients with follow-up from 1 to 235 months postdiagnosis were retrieved. Tumor HPV status was determined by HPV E6-targeted multiplex real-time PCR/p16 semiquantitative immunohistochemistry and cyclin D1 and pRb expression by semiquantitative immunohistochemistry. Determinants of recurrence and mortality hazards were modeled using Cox regression with censoring at dates of last followup. The HPV-positivity rate was 37% (91% type 16). HPV was a predictor of recurrence, an event (recurrence or death) and death after adjustment for clinicopathological variables. There were inverse relationships between HPV status and cyclin D1 and pRb. On univariate analysis, cyclin D1 predicted locoregional recurrence, event and death and pRb predicted event and death. Within the HPV-positive group, after adjusting for clinicopathological factors, patients with cyclin D1-positive cancers had up to a eightfold increased risk of poor outcome relative to those with cyclin D1-negative tumors. However, within the HPV-negative group, there was only a very small adjusted increased risk. A combination of pRb and HPV did not provide additional prognostic information. Our data provide the first evidence that a combination of HPV and cyclin D1 provides more prognostic information in oropharyngeal cancer than HPV alone. If findings are confirmed, treatment based on HPV and cyclin D1 may improve outcomes.Insight into molecular pathways underpinning carcinogenesis has opened the way for a new generation of molecular markers to predict cancer outcome. There is increasing use of combinations of molecular prognostic markers in human cancer.1,2 Many squamous cell carcinomas of the oropharynx arise following disruption of the p53 and/or the retinoblastoma (pRb) pathways by mutagens present in tobacco or alcohol. However, it is now accepted that up to 60% of oropharyngeal cancers are induced by human papillomavirus (HPV). [3][4][5][6][7] In the HPV-induced subset, the p53 and pRb pathways are inactivated by the HPV E6 and E7 oncoproteins, respectively. HPV-induced cancers appear biologically distinct; most notable is their more favorable prognosis. 8,9 Retinoblastoma (pRb) pathway proteins (p16 ink4 , cyclin D1

E6 viral protein ratio correlates with outcomes in human papillomavirus related oropharyngeal cancer

“…The inverse relationship between HPV status and cyclin D1 has been well documented. 10 It was therefore noteworthy that in the current study, cyclin D1 was not a significant prognostic factor after adjusting for E6*I/E6*II ratio in addition to known prognostic variables.…”

“…10,12 EGFR positivity was defined as membrane staining of at least moderate intensity in 10% of cancer cells. 12 Cyclin D1 positivity was defined as nuclear staining of at least moderate intensity in 10% of cancer cells.…”

“…12 Cyclin D1 positivity was defined as nuclear staining of at least moderate intensity in 10% of cancer cells. 10 Detection of HPV E6*I and E6*II splice variants by quantitative RT-PCR Four to 8 sections of 10mm formalin fixed paraffin embedded samples were used for RNA extraction with RNeasy FFPE kit (Qiagen) according to manufacturer's instructions. RNA extracts were quantified and assessed using NanoDrop ND 1000 (Thermo Fisher Scientific, USA).…”

“…Others have reported similar findings. 10 EGFR is abnormally activated in approximately 80% of head and neck cancers. 11 EGFR expression has been associated with poorer prognosis in head and neck cancer, although previous study indicated that the effect of EGFR on outcome in OSCC may be limited to HPV negative OSCC.…”

Human papillomavirus, smoking status and outcomes in tonsillar squamous cell carcinoma

“…p16 protein expression was determined by semiquantitative immunohistochemistry using the JC2 p16 antibody (Neomarkers, USA) as previously described. 23 Staining was strong and diffuse and was essentially all or none. Weak focal staining was recorded as negative.…”