[Analysis of human papillomavirus type 16 E6/E7 mRNA in cervical cancers and precancerous lesions by means of the polymerase chain reaction with reverse transcriptase reaction]

Abstract

We analyzed HPV-16 E6/E7 mRNA in human uterine cervical carcinomas and cervical intraepithelial neoplasias (CINs) by polymerase chain reaction (PCR) with reverse transcription (RT-PCR). Simultaneously total RNA and DNA were extracted from 6 cervical carcinomas, 14 CINs and 2 normal cervical tissues by the guanidium isothiocyanate/CsCl method. HPV-16 DNA was detected in 3 cervical carcinomas and 6 CINs. HPV-16 E6/E7 transcripts were detected in all HPV-16 DNA positive cervical carcinomas and CINs. In 2 cervical carcinomas and 5 CINs, 2 spliced E6 mRNA (E6*I and E6*II) and full length E6 mRNA were detected. In one cervical carcinoma and in one CIN, only full length E6-E7 mRNA was detected. Sequence analysis of cloned PCR products showed that both transcripts were generated by splicing out an intron in E6, from nucleotides (nt) 226 to 409 in E6*I and from nt 226 to 526 in E6*II. There was no significance difference in HPV-16 E6/E7 mRNA patterns between cervical carcinomas and CINs. This sensitive RT-PCR technique was available for analysis of HPV-16 mRNA in the small specimens.