食管癌凋亡通路中Survivin调控Bad的分子机制研究

我们最新研究结果显示,新疆哈萨克族食管癌发生发展重要的原癌基因Survivin，在小样本癌组织中的表达水平与Bad 呈负相关，并且在细胞中Bad 在转录水平随Survivin的变化而改变。但目前Survivin与Bad之间的关系尚不清楚。我们推测，Survivin可能作为转录调控因子，调控Bad的表达。Survivin未知的调控作用将会打破对Survivin传统的认识。因此，本项目拟采用RNA干扰、实时荧光定量PCR、Western blot、流式细胞技术等方法，在食管癌细胞株Eca109中，通过沉默Survivin检测Bad的mRNA与蛋白表达水平以及细胞凋亡状态；并运用ChIP方法寻找Survivin调控Bad的证据。同时，在哈萨克族食管癌组织标本中确认Bad和Survivin的相关性。从而为丰富Survivin在肿瘤发生发展过程中调控细胞凋亡新的分子生物学机制提供更详细的理论依据。

Our new results showed that mRNA expression of Bad was negatively correlated with Survivin in cancer tissues from small samples,and expression of Bad in mRNA was changed with Survivin, the proto-oncogene survivin which plays key role in occurrence and development of esophageal cancer in Kazak in Xinjiang. However, the relationship between Survivin and Bad is unclear. We speculate that Survivin may be as a transcription factor, could regulate the expression of Bad. This unknown function will change the traditional understanding of Survivin role.So,in this study,Sh-RNA targeting the Survivin gene will be transfected into ES Eca109 cells.After transfection,Bad expression will be detected by real-time PCR and Western blot, and apoptosis will be examined by flow cytometry.Furthermore, ChIP will be used to find the evidence that Survivin regulate Bad in transcriptional level.Meanwhile, detect tissue samples to confirm the correlation between Bad and Survivin.It will provide a more detail on the regulation of apoptosis in tumor occurrence and development and enrich molecular biological mechanism.