The causative role of high-risk human papillomavirus (HR-HPV) in cervical cancer development is well recognized, but HPV infection was less common in cervical adenocarcinoma(CADC) than squamous cell carcinoma(SCC) and CADC is diverse pathologically and in HPV status. Nevertheless, most studies to date have focused primarily on viral DNA rather than the viral transcription. The aim of this study was to investigate the presence of HR-HPV in cervical cancer tissues at HPV DNA level and HPV oncogenes mRNA level by polymerase chain reaction(PCR) and in situ hybridization (ISH) respectively.
We studied DNA and mRNA levels of HPV in paraffin-embedded samples from patients with CADC and SCC. 60 cases of CADC and 14 cases of SCC were included. Cases were tested for HPV using whole-tissue sections (WTS) and laser-capture microdissection(LCM). All cases were HPV-tested by L1 based broad-spectrum SPF10-DEIA-LiPA25 PCR. HR-HPV mRNA was assayed by novel RNAscope ISH. Type-specific oligonucleotide probes were used for the RNA detection of HPV 16,18,26,31,33,35,39,45,51,52,53,56,58,59,66,68,73 and 82.
HPV DNA was detected in all 14 (100.0%) SCC and in 36 out of 60 (60.0%) CADC cases by WTS-PCR. Overall, the HR-HPV mRNAs was detected in 12 out of 14(85.7%) SCC and 20 out of 60(33.3%) CADC by RNAscope ISH. 20 out of 36 (55.6%) WTS-PCR HPV DNA+ CADC cases detected HR-HPV mRNAs. The remaining 24 (100%) cases of WTS-PCR HPV DNA- CADC were also HPV mRNA-. Also, 16 out of 36 cases of WTS-PCR HPV DNA+ with multiple HPV infections were tested by LCM-PCR to determine whether one or more viruses are present in one lesion. 11 out of 16 (68.8%) multiple HPV infection cases were LCM-PCR HPV DNA- and were HPV mRNA-; 4 out of 16(25%) were LCM-PCR HPV DNA+ and HPV mRNA+; Only 1 case was LCM-PCR HPV DNA+ and HPV mRNA-. In CADC, the kappa coefficient of RNAscope and WTS-PCR was 0.500 (P＜0.001), while the kappa coefficient of RNA scope and LCM-PCR was 0.846 (P=0.001).
| HPV DNA / HPV mRNA | RNAscope (HPV mRNA) | ||
| Positive | Negative | ||
| WTS-PCR (HPV DNA) | Positive | 20 | 16 |
| Negative | 0 | 24 | |
| Kappa coefficient | 0.500 (P＜0.001) | ||
Compared with WTS-PCR, using LCM-PCR for HPV DNA evaluation yielded lower prevalence of HPV DNA and better concordance of HPV mRNA and DNA when using RNAscope assay to evaluate the expression of HPV mRNA. Overall, HR-HPVs exist in CADC tissue with less active transcription, which implies that the causal role of HPV in CADC development need further study.