Maesawa C, Tamura G, Suzuki Y, Ishida K, Saito K, Satodate R
Department of Pathology, Iwate Medical University School of Medicine, Morioka.
Jpn J Cancer Res. 1992 Dec;83(12):1253-6. doi: 10.1111/j.1349-7006.1992.tb02754.x.
For the rapid and sensitive detection of p53 gene mutations in esophageal endoscopic biopsy specimens, we combined cell sorting with the polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) analysis. Mutations in exons 5-8 of the p53 gene were investigated by PCR-SSCP analysis using 10(3) sorted nuclei obtained from each endoscopic biopsy specimen of 16 patients with esophageal cancer. DNAs extracted from their respective surgical specimens were investigated by a conventional method of PCR-SSCP analysis. Mutations in the biopsy specimens were detected in 6 of the 12 aneuploid tumors but in none of the 4 diploid tumors. After tumor cell enrichment by cell sorting, one mutation in exon 8 became apparent, which could not be detected from the surgical specimen by a conventional method of PCR-SSCP analysis. This method should improve the sensitivity of detecting p53 gene mutations, and provides additional information concerning the DNA ploidy pattern in the tumors.
为了快速、灵敏地检测食管内镜活检标本中的p53基因突变，我们将细胞分选与聚合酶链反应及单链构象多态性（PCR-SSCP）分析相结合。采用PCR-SSCP分析，对16例食管癌患者的每例内镜活检标本中获取的10³个分选核进行检测，研究p53基因第5至8外显子的突变情况。用传统的PCR-SSCP分析方法对从其各自手术标本中提取的DNA进行检测。在12例非整倍体肿瘤的活检标本中检测到6例有突变，而4例二倍体肿瘤中均未检测到突变。通过细胞分选使肿瘤细胞富集后，第8外显子的一个突变变得明显，而用传统的PCR-SSCP分析方法从手术标本中无法检测到该突变。该方法应能提高检测p53基因突变的灵敏度，并提供有关肿瘤中DNA倍体模式的额外信息。
