胰腺占位性病灶细针穿刺活检标本中ppENK和TFPI-2基因甲基化检测的可行性及其应用

The feasibility and application of the methylation status detection of ppENK gene and TFPI-2 gene in the pancreatic space occupying lesions tissue strip obtained by EUS-FNA

目的检测胰腺组织ppENK和TFPI-2基因甲基化状态,探讨胰腺占位性病灶细针穿刺活检标本中ppENK和TFPI-2基因甲基化检测的可行性及其应用。方法采用重亚硫酸盐测序PCR(BSP)法检测86例胰腺占位性病变组织中ppENK和TFPI-2基因启动子区CPG岛的甲基化状态,采用单因素及多因素回归Logistic回归分析ppENK基因和TFPI基因甲基化检测的诊断价值,以及EUS-FNA联合基因甲基化检测在胰腺占位性病变中的诊断意义。结果本研究86例病例中,甲基化成功率为91.9%(79/86),TFPI-2和ppENK基因的甲基化检测率分别为95.9%(75/79)和91.1%(72/79)。肿瘤组与炎症组以及肿瘤组和对照组的甲基化率差异均有统计学意义(P<0.05),而炎症组与对照组甲基化率差异无统计学意义。受试者工作特征曲线结果显示:TFPI-2基因、ppENK基因甲基化水平诊断胰腺癌的敏感度分别为83.6%、74.6%,特异度分别为81.8%、86.4%,诊断率分别为89.7%、81.2%,以TFPI-2基因或ppENK基因甲基化水平诊断胰腺癌的敏感度为92.5%,特异度为79.8%,而以基因TFPI-2和ppENK甲基化水平进行诊断,其敏感度为74.6%,特异度为91.7%。结论TFPI-2和ppENK基因在胰腺肿瘤组织中具有高甲基化水平,且具有良好的敏感性和特异性,是胰腺癌的有利诊断指标,结合EUS-FNA,能够有效提高胰腺占位性病变的术前诊断。

Objective To detect the methylation status of ppENK gene and TFPI-2 gene in pancreatic tissue, and to explore the feasibility and application of the methylation status detection of ppENK gene and TFPI-2 gene in the pancreatic space occupying lesions obtained by EUS-FNA. Methods The methylation status of CPG islands in the promoter region of ppENK gene and TFPI-2 gene in 86 cases pancreatic cancer tissues and adjacent tissues were detected by bisulfite genomic sequencing PCR（BSP） ,and the clinical pathological characteristics between pancreatic cancer and adjacent tissues were analyzed by Logistic. Results In this study of 86 cases, the success rate of methylation was 91.9% （79/86）. The detection rates of methylation of TFPI-2 gene and ppENK gene genes were 95.9% （75/79） and 91.1% and the normal control and the normal control PI-2 gene and ppENK （72/79）, respectively. There were significantly statistic differences between the tumor group group （ P 〈 0.05 ） , while there was no significant difference between the inflammatory group group. The results of receiver operating characteristic curve showed that the sensitivity of TF- gene methylation level in the diagnosis of pancreatic space occupying lesions were 83.6% and 74.6% ,respectively,and the specificity was 81.8% and 86. 4% respectively, diagnosis rate was 89.7% and 81.2% respectively. The diagnosis of pancreatic space occupying lesions sensitivity of the methylation level of TF- PI-2 gene or ppENK gene was 92.5%, the specificity was 79.8%, and the sensitivity of the TFPI-2 gene and ppENK gene methylation level of diagnosis of pancreatic space occupying lesions was 74.6% , the specificity was 91.7%. Conclusion TFPI-2 and ppENK genes have high methylation level in pancreatic neoplasms and have good sensitivity and specificity. It is a useful diagnostic index for pancreatic cancer. Combined with EUS-FNA, it can effectively improve the preoperative diagnosis.