Decreased PADI4 mRNA Association with Global Hypomethylation in Hepatocellular Carcinoma During HBV Exposure
Decreased PADI4 mRNA Association with Global Hypomethylation in Hepatocellular Carcinoma During HBV Exposure
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DOI：
10.1007/s12013-012-9417-3
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发表时间：
2013-03
影响因子：
2.6
通讯作者：
Changsong Zhang;L. Fan;T. Fan;Dong-hao Wu;Lu Gao;Y. Ling;Jing Zhu;Rong Li;Lixin Wei
中科院分区：
文献类型：
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作者：
Changsong Zhang;L. Fan;T. Fan;Dong-hao Wu;Lu Gao;Y. Ling;Jing Zhu;Rong Li;Lixin Wei
为了深入了解肽基精氨酸脱亚胺酶4型（PADI 4）的作用，我们确定了在肝癌发生过程中HBV暴露期间PADI 4 mRNA和整体低甲基化之间的关系。我们通过MSP分析了74例HCC中Line-1甲基化，通过实时PCR分析了CD 133 mRNA。用5-Aza-CdR和TSA处理HCC癌系（7721、Huh 7和Hep-G2）。HCC组织中的PADI 4 mRNA（平均− Ct= 1.41）低于非HCC组织中的PADI 4 mRNA（平均− Ct= 3.10）。74例HCC患者中仅20例（27%）PADI 4表达升高，但54例（73%）患者PADI 4表达降低。HCC患者中PADI 4 mRNA的下降与Line-1去甲基化显著相关。PADI 4 mRNA在1系HCC患者中的表达低于1系HCC患者，1系HCC MI <−0.15（平均− AMI Ct= −2.66），而1系HCC患者中的PADI 4 mRNA表达高于1系HCC患者，1系HCC MI >= −0.15（平均− AMI Ct= −1.02）。结果表明，显示Line-1低甲基化的HCC被认为是沉默PADI 4 mRNA。HBV >= 105（copy/ml）的HCC患者PADI 4 mRNA表达水平低于HBV < 105（copy/ml）的HCC患者。5-Aza-CdR和TSA处理后，发现TSA能显著诱导Huh 7和Hep-G2细胞中PADI 4 mRNA的表达，但对7721细胞无明显影响。同时，5-Aza-CdR与TSA联合作用可诱导肝癌细胞PADI 4 mRNA的表达，而单独作用对PADI 4 mRNA表达无影响。结果表明，在HBV暴露期间，HCC中PADI 4 mRNA的降低与整体低甲基化相关。
To gain insight into the role of peptidylarginine deiminase type 4 (PADI4), we determined the relationship between PADI4 mRNA and global hypomethylation during HBV exposure in hepatocarcinogenesis. We analyzed Line-1 methylation by MSP, and CD133 mRNA by real-time PCR in 74 HCC. The HCC cancer lines (7721, Huh7, and Hep-G2) were treated with 5-Aza-CdR and TSA. PADI4 mRNA were lower in HCC tissues (Mean−∆Ct= 1.41) than that in Non-Hcc tissues (Mean−∆Ct= 3.10). Expression of PADI4 was elevated in only 20 (27 %) of the 74 HCC patients but decreased in 54 (73 %) of the patients. The declined PADI4 mRNA was significantly associated with Line-1 demethylation in HCC patients. PADI4 mRNA were lower in HCC patients with Line-1 ∆MI <−0.15 (Mean−∆∆Ct= −2.66) than those with Line-1 ∆MI >= −0.15 (Mean−∆∆Ct= −1.02). The results suggested that HCC showing hypomethylation of Line-1 is considered to be silencing PADI4 mRNA. And the lower PADI4 mRNA was also found in HCC patients with HBV >= 105(copy/ml) than those with HBV < 105(copy/ml). After treated by 5-Aza-CdR and TSA, we found that PADI4 mRNA induced significantly by TSA in Huh7 and Hep-G2 cells, but not in 7721 cells. Meanwhile, PADI4 mRNA induced by the combination of 5-Aza-CdR and TSA in HCC cells, and it could no effect for exposure to 5-Aza-CdR alone. The results suggested that decreased PADI4 mRNA is associated with global hypomethylation in HCC during HBV exposure.