Detection of high-risk cervical intraepithelial neoplasia and cervical cancer by amplification of transcripts derived from integrated papillomavirus oncogenes

Cervical cancer emerges from cervical intraepithelial neoplasia (CIN) induc ed by high-risk RPV (HR-HPV) infections, However, the vast majority of CIN lesions regresses spontaneously, and only a few lesions persist or progress to invasive carcinoma. On the basis of morphological criteria, it is not p ossible to differentiate high-grade lesions that will regress or persist fr om those that inevitably will progress to invasive cancers. In most cervica l carcinomas, human papillomavirus (HPV) genomes are integrated into host c ell chromosomes and transcribed into mRNAs encompassing viral and cellular sequences. In contrast, in early preneoplastic lesions, NPV genomes persist as episomes, and derived transcripts contain exclusively viral sequences. Thus, detection of HPV transcripts derived from integrated NPV genomes may specifically indicate both CIN lesions at high risk for progression as well as invasive cervical cancers, Here, we established a protocol for the ampl ification of papillomavirus oncogene transcripts (APOT) from cervical speci mens that allows us to distinguish episome- from integrate-derived HPV mRNA s, Cervical swab and biopsy samples from. 549 patients attending outpatient clinics for cervical dysplasia were screened for the presence of HPV DNA, and 155 samples that were positive for either HPV type 16 (n = 143) or 18 ( n = 12) sere subjected to the APOT assay. In samples derived from normal ce rvical epithelia (n = 19) or low-grade cervical lesions (CIN I, n = 10), no integrate-derived HPV transcripts were found. In contrast, in 1 (5%) of 22 samples derived from CIN II lesions, in 10 (16%) of 64 samples from patien ts with CIN III lesions, and in 35 (88%) of 40 samples from patients with c ervical cancer, integrate-derived HPV transcripts were detected. Thus, dete ction of integrate-derived HPV transcripts in cervical swabs or biopsy spec imens by the APOT assay points to advanced dysplasia or invasive cervical c ancer.