乳腺癌Jab1蛋白调控Nov基因的表达受甲基化水平影响

Jab1 Regulating Nov Affected by DNA Methylation in Breast Cancer Cell Line

为探讨癌基因Jab1在乳腺癌发生发展中的功能,首先在乳腺癌细胞MDA231中建立了慢病毒介导的Jab1基因的表达干扰系统,并通过人基因表达谱芯片分析结果以及实时定量PCR实验筛选出受Jab1调控的下游基因;此外,通过实时定量PCR以及Western blot实验证实Jab1干扰表达能降低Nov基因在转录和翻译水平表达,而且Nov启动子区域存在4个高甲基化CpG位点.进一步使用甲基转移酶抑制剂处理Jab1干扰表达细胞,发现与未使用抑制剂处理细胞相比,Nov基因的mRNA和蛋白质表达水平发生明显上调,说明Jab1基因调控Nov表达受甲基化水平的影响,提示Jab1可能是通过表观遗传学水平调控Nov基因的表达.

In order to study the effects of Jab1 in breast cancer,aJab1 gene knockdown system in breast cancer cell line MDA231 was established with lentivirus mediated RNA interference.Then our study confirmed that Nov gene was regulated by Jab1 through human expression microarray and Real-Time PCR.Moreover,the knockdown of Jab1 decreased the expression level of mRNA and protein of Novgene.Moreover,4CpG sites of Novpromotor have high methylation rates.The expression level of Novgene had no significant difference after adding methyltransferase inhibitor 5′-aza-dC in Jab1 knockdown cell lines comparing to the expression of Nov down regulated in Jab1 knockdown cell lines without inhibitor which indicats that Jab1 may regulate Novgene through epigenetics.