摘要
目的探讨甲基化酶抑制剂5-氮杂-2L脱氧胞苷（5-AzadC）及组蛋白去乙酰化酶抑制剂曲古菌素A（TSA）对胰腺癌Panc-1细胞系中TFPI-2基因甲基化水平及基因表达的影响。方法单独或联合应用5-Aza—dC及TSA处理Panc-1细胞，应用甲基化特异性聚合酶链反应、逆转录聚合酶链反应及蛋白印迹实验检测药物处理前后Panc-1细胞TFPI-2基因启动子区甲基化状态、mRNA及蛋白表达情况。结果经5-Aza-dC单独处理或5-Aza—dC及TSA联合处理Panc-1细胞后，TFPI-2基因启动子区高甲基化状态得到逆转，表现为非甲基化，原本不表达的TFPI-2基因mRNA及蛋白重新表达，两组作用效果相似。经TSA单独处理的Panc-1细胞TFPI-2基因启动子区仍为异常高甲基化状态，原本不表达的TFPI2基因未见重新表达。结论胰腺癌Panc1细胞系中TFPI-2基凶启动子区高甲基化可能是导致该基因失活的主要原因，5Aza—dC单独作用或与TSA联合作用均能逆转TFPI-2基因的高甲基化状态，使该基因重新表达。
Objective To investigate effects of 5 aza-2＇-deoxycytidine （5-Aza dC） and trichosta- tin A （TSA） on the methylation and expression of TFPI-2 gene in pancreatic cancer Panc-1 cells. Methods We treated pancreatic cancer Pane-1 cells with 5-Aza-dC and TSA alone or combined. TFPI- 2 gene＇s DNA, mRNA and protein were determined by MSP, RT-PCR and Western blot. Results The hypermethylation of TFPI-2 gene in Pane-1 ceils was reversed after treated with 5-Aza-dC alone or in combination with TSA. The re-introduction or raised expression of TFPI-2 mRNA and protein was detected in Pane-1 cells after treated with 5 Aza-dC alone or in combination. However, we did not see the hypermethylation of TFPI-2 gene reversed nor detected TFPI-2 gene mRNA and protein after treated with TSA alone. Conclusions The methylation of the promoter region for the TFPI-2 gene is a main cause for its transcriptional activation in Pane-1 cells. The hypermethylation of TFPI-2 gene in Panc 1 cell was reversed and the expression of TFPI-2 gene mRNA and protein was detected after trea ted with S-Aza-dC alone or in combination with TSA. Treatment of TSA alone does not have significant effect on TFPI-2 gene reexpression.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2012年第3期226-229,共4页
Chinese Journal of Hepatobiliary Surgery
基金
基金项目：国家自然科学基金（81071734）
关键词
胰腺肿瘤
基因表达
曲古菌素A
Pancreatic neoplasms
Gene expression
Trichostatin A
