【作者】 董桂银；

【导师】 钱叶本； 朱立新； 耿小平；

【作者基本信息】 安徽医科大学 ， 外科学， 2006， 硕士

【摘要】 目的：了解肝细胞癌(hepatocellular carcinoma，HCC)中SLIT2、BLU、ppENK和TMS1基因的甲基化状态及临床意义。方法：采用甲基化特异性聚合酶链反应(methylation-specific PCR，MSP)技术，检测50例HCC癌组织及相应的癌旁组织，4个HCC细胞株，8例肝血管瘤旁和4例供肝组织，了解SLIT2、BLU、ppENK和TMS1基因在上述组织中的甲基化状态，并分析其与临床资料间的相关性。结果：4例供肝组织均未发生甲基化，8例肝血管瘤旁组织有1例发生BLU甲基化，4个HCC细胞株分别有3、3、1和2个发生SLIT2、BLU、ppENK和TMS1甲基化，50例HCC癌组织及其相应的癌旁组织中，分别检测到39例(78％)、26例(52％)发生SLIT2甲基化，33例(66％)、18例(36％)发生BLU甲基化，14例(28％)、8例(16％)发生ppENK甲基化，14例(28％)、7例(14％)发生TMS1甲基化。癌组织与癌旁组织甲基化率比较，SLIT2、BLU差异有显著性(χ~2=7.429，P=0.006；χ~2=9.004，P=0.003)，ppENK、TMS1差异无显著性(χ~2=2.008，P=0.148；χ~2=2.954，P=0.086)。100％的癌和78％的癌旁组织至少有1个基因发生了甲基化。癌组织中只含1种基因甲基化的标本为15份(30％)，含2种基因甲基化的标本为20份(40％)，3种基因甲基化的标本为15份(30％)，4种基因甲基化的标本为0份。癌组织与癌旁组织SLIT2、BLU、ppENK和TMS1基因甲基化发生情况之间无统计学相关性(P＞0.05)。TNM Ⅲ期患者癌组织SLIT2甲基化率高于TNM Ⅰ／Ⅱ期患者(χ~2=4.432，P=0.035)。肝功能Child B级患者癌组织BLU甲基化率高于Child A级患者(χ~2=4.193，P=0.041)。癌组织BLU甲基化阳性患者的肿瘤直径大于BLU甲基化阴性患者(t=3.038，P=0.004)。癌旁组织BLU甲基化阴性患者的血清AFP水平高于BLU甲基化阳性者(t=2.244，P=0.029)。癌组织BLU甲基化阳性患者更多还原

【Abstract】 Objective: To investigate methylation of SLIT2, BLU, ppENK and TMS1 genes in hepatocellular carcinoma. Methods: Methylation-specific PCR was adopted to investigate methylation of SLIT2, BLU, ppENK and TMS1 genes in 50 cases of HCC and corresponding non-tumor liver tissue, 4 HCC cell lines, 8 normal liver tissue in liver angioma and 4 liver donors. The relationship between methylation of SLIT2, BLU, ppENK and TMS1 genes and clinical data was analyzed. Results: 4 liver donors have no methylation of these genes. 1 of 8 normal liver tissue in liver angioma was detected methylation in BLU. No normal liver tissue in liver angioma was detected methylation in SLIT2, ppENK and TMS1. Methylation of SLIT2, BLU, ppENK and TMS1 was found in 3, 3, 1 and 2 of 4 HCC cell lines, respectively. In 50 cases of HCC and corresponding non-tumor liver tissue, the frequency of SLIT2 methylation was 78%, 52%, respectively; the frequency of BLU methylation was 66%, 38%, respectively; the frequency of ppENK methylation was 28%, 16%, respectively; the frequency of TMS1 methylation was 28%, 14%, respectively; Methylation of SLIT2 and BLU was more frequent in HCC than in non-tumor liver tissue(x~2=7.429, P=0.006; x~2=9.004, P=0.003). There was no significant difference between methylation of ppENK and TMS 1 in HCC and non-tumor liver tissue(x~2=2.098, P=0.148; x~2=2.954, P=0.086). At least one gene was methylated in 100% of HCC and 78% of non-tumor liver tissue. One gene was methylated in 15 of HCC (30%) . Two genes were methylated in 20 of HCC (40%) . Three genes were methylated in 15 of HCC (30%). No cases had methylation of 4 genes.更多还原