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Analysis of Tumor-Related Genes In Pancreatic Juice Aiming to be Useful for Early Diagnosis of Pancreatic Cancer

胰液中肿瘤相关基因的分析有助于胰腺癌的早期诊断

基本信息

批准号：
14370177
负责人：
SAWABU Norio
金额：
$ 3.9万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

We analyzed mutations of p53 in DNA extract not only from the sediment(Sed) but also from the supernatant(sup) of aspirated pancreatic juice(PJ) by ERCP and bile obtained by PTC or ERBD. The incidence of p53 mutations was 43%(9 of 21) and 29%(6 of 21) in the Sup and the Sed of PJ from patients with pancreatic cancer(PCa), respectively, and nurse to 52%(11 of 21) in the combination assay with Sup and Sed. Moreover, p53 mutations were detected in 47%(7 of 15) of PCa cases in which the cyto logic diagnosis was negative. Among 25 patients with chronic pancreatitis(CP), none harbored mutant p53. In bile abtained from patients with biliary tract cancer, the positive rate of p53 mutations was 50%(15 of 30) in the Sup and 33%(10 of 30) in the Sed. On the other hand, there were no pantients with cholelithiasis harboring mutant p53. These results could indicate that the incidence of p53 mutation was higher in the Sup than in the Sed, and the specificity of p53 mutation was higher in the Sup than … More in the Sed, and the specificity of p53 mutation for cancer was very high. Mesothlin mRNA expression in the PJ was discovered in 11(52%) of 21 patients with PCa,5(45%) of 11 with intraductal papillary mucinous neoplasm(IPMN) and 3(14%) of 22 with CP. There was significant difference between PCa and CP for mesothelin mRNA. We developed an highly sensitive quantitative analysis for h-TERT mRNA using real-time RT-PCR and measured the amount of h-TERT mRNA in serum and PJ. The levels of h-TERT mRNA were elevated in almost all cases with PCa but also in considerable cases with benign lesions, suggesting low specificity of h-TERT mRNA for cancer. h-TERT mRNA in body fluids from benign lesions originates from lymphocyte.Using methylation-specific PCR, we nextly analyzed the methylation status of 3 CpG islands of ppENK, SARP2 and NPTX2 in PJ samples. Aberrant methylation of ppENK in the PJ was detected in 14(50%) of 28 patients with PCa, 4(27%) of 15 with IPMN and 1(15%) of 20 with CP, and that of NPTX2 was detected in 24(77%) of 35 patients with PCa, 10(63%) of 16 with IPMN and 6(43%) of 13 with CP. The incidence of aberrant methylation of ppENK or NPTX2 was significantly higher in PCa than in CP. Furthermore, we found aberrant methylation of SARP2 in PJ in 24(74%) of 35 patients with PCa, 15(94%) of 16 with IPMN and 2(11%) of 19 with CP. The incidence of aberrant methylation of SARP2 was significantly higher in PCa or IPMN than in CP. Although the prevalence of aberrant methylation of the tumor-related genes in PJ from PCa was strikingly high, its specificity for cancer was not sufficient. There was no significant difference between benign and malignant groups of IPMN, suggesting that aberrant menthylation of these genes is a early event in PCa development, and quantitative detection of aberrant methylation in PJ may have potential diagnostic implication for PCa. Less

我们不仅分析了沉积物（Sed）DNA提取物中的p53突变，还分析了ERCP吸出的胰液（PJ）的上清液（sup）以及PTC或ERBD获得的胆汁中的DNA提取物中的p53突变。胰腺癌（PCa）患者 PJ 的 Sup 和 Sed 中 p53 突变的发生率分别为 43%（21 例中的 9 例）和 29%（21 例中的 6 例），而在 Sup 和 Sed 联合检测中，p53 突变的发生率为 52%（21 例中的 11 例）。此外，47%（15 例中的 7 例）细胞学诊断为阴性的 PCa 病例中检测到 p53 突变。在25名慢性胰腺炎（CP）患者中，没有人携带p53突变体。在胆道癌患者胆汁中，Sup 中 p53 突变阳性率为 50%（30 例中的 15 例），Sed 中 p53 突变的阳性率为 33%（30 例中的 10 例）。另一方面，没有胆石症患者携带突变型p53。这些结果可以表明，Sup中p53突变的发生率高于Sed，并且Sup中p53突变的特异性高于Sed中，并且p53突变对癌症的特异性非常高。 21例PCa患者中11例（52％）发现PJ中间皮素mRNA表达，11例导管内乳头状粘液性肿瘤（IPMN）中5例（45％）发现，22例CP中3例（14％）发现PJ中间皮素mRNA表达。 PCa和CP之间的间皮素mRNA存在显着差异。我们使用实时 RT-PCR 开发了一种高度灵敏的 h-TERT mRNA 定量分析方法，并测量了血清和 PJ 中 h-TERT mRNA 的量。 h-TERT mRNA 水平在几乎所有 PCa 病例中均升高，但在相当多的良性病变病例中也升高，表明 h-TERT mRNA 对癌症的特异性较低。良性病变体液中的h-TERT mRNA源自淋巴细胞。接下来，我们利用甲基化特异性PCR分析了PJ样本中ppENK、SARP2和NPTX2 3个CpG岛的甲基化状态。 28 例 PCa 患者中 14 例（50%）检测到 PJ 异常甲基化，15 例 IPMN 患者中检测到 4 例（27%），20 例 CP 患者中 1 例（15%）检测到 NPTX2 异常甲基化；35 例 PCa 患者中 24 例（77%）检测到 NPTX2 异常甲基化，16 例 IPMN 患者中检测到 10 例（63%），16 例 IPMN 患者中检测到 6 例（43%）。 13 与CP。 PCa 中 ppENK 或 NPTX2 异常甲基化的发生率显着高于 CP。此外，我们发现 35 名 PCa 患者中的 24 名（74%）、16 名 IPMN 患者中的 15 名（94%）和 19 名 CP 患者中的 2 名（11%）存在 PJ 中 SARP2 的异常甲基化。 PCa 或 IPMN 中 SARP2 异常甲基化的发生率显着高于 CP。尽管 PCa 中 PJ 中肿瘤相关基因异常甲基化的发生率惊人地高，但其对癌症的特异性还不够。 IPMN良性组和恶性组之间没有显着差异，表明这些基因的异常甲基化是PCa发展的早期事件，定量检测PJ中的异常甲基化可能对PCa具有潜在的诊断意义。较少的