Viral E6 is overexpressed via high viral load in invasive cervical cancer with episomal HPV16

Abstract

Background: The integration of HR-HPV genome into host DNA is regarded as a key step for the development of cervical cancer. However, HR-HPV genome indeed exists as episome except for integrant. It may be alternative mechanisms in episome-associated carcinogenesis, although, by which HPV 16 episome induces cervical carcinogenesis is unclear now. Methods: Ninety-three invasive cervical cancer tissues with HPV16 positive were collected. Viral physical status was calculated from comparing E2 to E6-copies and detection of viral load was made with realtime-PCR using copy numbers of E6. HPV16 E6 mRNA transcript levels were measured by realtime-PCR. The methylation frequency of HPV16 promoter was detected by PCR and pyrosequencing. Results: In 93 samples, 21.5% (20/93) presented purely integrated viral genome, 53.8% (50/93) mixed viral genome, and 24.7% (23/93) purely episomal viral genome. Mean E6 expression in samples with purely episomal viral genomes was 7.13-fold higher than that with purely integrated viral genomes. Meanwhile, viral load in samples with purely episomal viral genomes was 4.53-fold higher than that with purely integrated viral genomes. E6 mRNA expression increased with the viral load in purely episomal cases. There were no differences of mean methylation frequency between purely episomal and integrated virus and among five CpG positions of HPV16 promoter for all samples. And there also was no correlation between E6 mRNA expression and methylation of HPV16 promoter among all samples with purely HPV16 episomal virus. Conclusions: HPV16 with the purely episomal viral genomes exists in a definite proportion of invasive cervical cancer, and episomal HPV16 also overexpresses E6 mRNA, probably through a high level of viral load.