RNA-arbitrarily primed (AP)-PCR-SSCP法によるヒト肝細胞がん細胞株mRNA異常の検出の試み

概要

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To detect minute mRNA aberrations in human hepatocellular carcinoma (HCC), we utilized a new RNA fingerprinting method which combines RNA-arbitrarily primed-polymerase chain reaction (RNA-AP-PCR) and single-strand conformation polymorphism (SSCP) analysis. After total RNAs had been extracted from six human liver cell lines including four HCC cell lines, we performed RNA-AP-PCR-SSCP analysis with three primers. With this analysis, we detected numerous regions showing shifted bands (shifted regions) indicative of mRNA aberrations or polymorphisms. We selected four such regions and determined their nucleotide sequences. It was revealed that shifted region P represented a silent mutation of the HCC cell line Hep 3B at codon 65 of human ribosomal protein S20 CDNA (GEN-L06498) and that shifted regions Q, R and S represented single base deletions at the 3' end of the human fetal brain CDNA (GEN-420F08) of the cell line Hep 3B suggesting associated mRNA aberrations. Thus, RNA-AP-PCR-SSCP analysis is useful for unbiased detection of minute aberrations in human HCC mRNA, including single-base substitutions which are not detectable by RNA-AP-PCR analysis alone.
東京女子医科大学の論文
1996-10-25