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

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  • NISHIKAWA,Akira
    Department of Obstetrics and Gynecology, Sapporo Medical College
  • SHIMADA,Masamitsu
    Department of Molecular Biology, Cancer Research Institute, Sapporo Medical College:Bio Research Laboratories, Takara Shuzo Co. Ltd.
  • FUKUSHIMA,Michio
    Department of Obstetrics and Gynecology, Sapporo Medical College:Department of Molecular Biology, Cancer Research Institute, Sapporo Medical College

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  • 逆転写酵素を利用したPolymerase Chain Reaction法 (RT-PCR法) による子宮頚癌及びその前癌病変におけるヒトパピローマウイルス16型E6/E7 mRNAの解析

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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.

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