Detection of p53 Gene Mutations in the Supernatant of Pancreatic Juice and Plasma from Patients with Pancreatic Carcinomas
Detection of p53 Gene Mutations in the Supernatant of Pancreatic Juice and Plasma from Patients with Pancreatic Carcinomas
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DOI：
10.1097/00006676-200401000-00002
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发表时间：
2004-01
期刊：
影响因子：
2.9
通讯作者：
Ying Wang;Y. Yamaguchi;Hiroyuki Watanabe;K. Ohtsubo;Y. Motoo;N. Sawabu
中科院分区：
文献类型：
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作者：
Ying Wang;Y. Yamaguchi;Hiroyuki Watanabe;K. Ohtsubo;Y. Motoo;N. Sawabu
目的 纯胰液（PPJ）细胞学检查诊断胰腺癌（PCa）的敏感性仍然较低。由于敏感性不足或假阳性，PPJ 遗传分析的实用性似乎受到限制。为了改进 PCa 的分子诊断，我们分析了从沉积物和 PPJ 样品上清液中提取的 DNA 中 p53 和 K-ras 的突变。方法采用聚合酶链反应-单链构象多态性和直接测序技术分析p53外显子5-8突变。通过突变等位基因特异性扩增来检查密码子 12 处的 K-ras 突变。结果 PCa 患者的 PPJ 上清液中，分别有 42.9%（21 例中的 9 例）和 81.0%（21 例中的 17 例）检测到 p53 和 K-ras 突变。沉积物中p53和K-ras突变的发生率分别为28.6%和71.4%。通过上清液和沉淀物的联合检测，52.4%（21 例中的 11 例）PCa 病例中检测到 p53 突变。此外，在细胞学诊断阴性的 15 例 PCa 病例中，有 7 例 (46.7%) 检测到 p53 突变。在 25 名慢性胰腺炎 (CP) 患者中，没有人携带 p53 突变体，尽管在上清液中检测到 K-ras 突变的发生率为 28%（25 人中的 7 人），在沉积物中检测到的发生率为 20%（25 人中的 5 人）。此外，在 11 名 PCa 患者中，有 2 名在 PPJ 中检测到 p53 突变，其中 2 名在血浆中检测到 p53 突变带。结论 这些结果表明，通过在 PCa 的 PPJ 样本中使用 Sup 可以提高癌症特异性高的 p53 突变检测的灵敏度。 p53 的遗传分析可以补充 PPJ 细胞学。在血浆样本中的 PCa 中可检测到 p53 突变。
Aim The sensitivity of pure pancreatic juice (PPJ) cytology for the diagnosis of pancreatic carcinoma (PCa) is still low. The usefulness of genetic analyses of PPJ seems to be limited because of insufficient sensitivity or false positivity. To improve the molecular diagnosis of PCa, we analyzed mutations of p53 together with K-ras in DNA extracted not only from the sediment but also from the supernatant of PPJ samples. Method Polymerase chain reaction–single-strand conformation polymorphism and direct sequencing were used for analyses of p53 mutations in exons 5–8. K-ras mutations at codon 12 were examined by mutant allele-specific amplification. Results In PPJ supernatant from patients with PCa, p53 and K-ras mutations were detected in 42.9% (9 of 21) and 81.0% (17 of 21) of cases, respectively. The incidence of p53 and K-ras mutations in the sediment was 28.6% and 71.4%, respectively. By a combination assay with supernatant and sediment, p53 mutations were detected in 52.4% (11 of 21) of PCa cases. Moreover, p53 mutations were detected in 7 of 15 (46.7%) cases of PCa in which the cytologic diagnosis was negative. Among 25 patients with chronic pancreatitis (CP), none harbored mutant p53, although K-ras mutations were detected at an incidence of 28% (7 of 25) in the supernatant and 20% (5 of 25) in the sediment. In addition, mutant bands of p53 in plasma were detected in 2 of 11 patients with PCa in whom p53 mutations were detectable in PPJ. Conclusion These results suggest that the sensitivity of detection for p53 mutations with high cancer specificity could be improved by using the Sup in PPJ samples of PCa. Genetic analysis of p53 could complement PPJ cytology. p53 mutations were detectable in PCa from plasma samples.