论文部分内容阅读
目的研究子宫内膜癌(EC)中相关抑癌基因(SOCS-1、SOCS-3、3-OST-2、DLC-1)的启动子甲基化状态,探讨其临床病理意义。方法收集60例EC、79例增生子宫内膜和27例正常子宫内膜标本,运用甲基化特异性PCR(MSP)技术检测4个抑癌基因的甲基化状态;MSP和实时荧光定量PCR法检测表观遗传学药物处理前后Ishikaw a细胞株中抑癌基因甲基化及mRNA表达变化。结果在EC中,SOCS-1和DLC-1基因甲基化率较低(分别为13.3%和21.7%),SOCS-3和3-OST-2基因甲基化率很高(分别为88.3%和78.3%),且SOCS-3在复杂和不典型增生中就出现高频率的甲基化(分别为53.3%和54.2%)。3-OST-2甲基化与肿瘤分化呈正相关(P<0.05)。SOCS-3和3-OST-2在Ishikaw a细胞株中均呈完全甲基化状态,DNA甲基转移酶(DNMT)抑制剂5-Aza-dC或组蛋白去乙酰化酶(HDAC)抑制剂TSA处理后甲基化均得到部分逆转,mRNA表达明显增加,且TSA效果优于5-Aza-dC;两药联合处理后甲基化被完全逆转,mRNA水平显著升高。结论 SOCS-3、3-OST-2基因启动子甲基化在EC中是一个频发事件,且SOCS-3甲基化的发生早于3-OST-2。DNA甲基化和组蛋白修饰共同参与了3-OST-2和SOCS-3的基因表达调控,并存在交叉作用。
Objective To investigate the promoter methylation status of related tumor suppressor genes (SOCS-1, SOCS-3,3-OST-2 and DLC-1) in endometrial carcinoma (EC) and to explore its clinicopathological significance. Methods 60 cases of EC, 79 cases of proliferative endometrium and 27 cases of normal endometrium were collected. Methylation-specific PCR (MSP) was used to detect the methylation status of the four tumor suppressor genes. MSP and real-time fluorescence quantitative PCR Methylation and mRNA expression of tumor suppressor gene in Ishikaw a cell line before and after epigenetic drug treatment. Results The methylation rates of SOCS-1 and DLC-1 genes were lower in ECs (13.3% and 21.7%, respectively), while the methylation rates of SOCS-3 and 3-OST-2 genes were higher (88.3% And 78.3% respectively), and SOCS-3 showed high frequency of methylation (53.3% and 54.2%, respectively) in complex and atypical hyperplasia. Methylation of 3-OST-2 was positively correlated with tumor differentiation (P <0.05). Both SOCS-3 and 3-OST-2 were completely methylated in Ishikawa a cell line, and DNA methyltransferase (DNMT) inhibitor 5-Aza-dC or histone deacetylase (HDAC) After TSA treatment methylation was partially reversed, mRNA expression was significantly increased, and TSA better than 5-Aza-dC; methylation was completely reversed after treatment with the two drugs, mRNA levels were significantly increased. Conclusion The promoter methylation of SOCS-3,3-OST-2 gene is a frequent event in EC, and SOCS-3 methylation occurs earlier than that of 3-OST-2. DNA methylation and histone modifications are involved in the regulation of gene expression of 3-OST-2 and SOCS-3, with cross-talk.