Hamada Shin, Satoh Kennichi, Hirota Morihisa, Fujibuchi Wataru, Kanno Atsushi, Umino Jun, Ito Hiromichi, Satoh Akihiko, Kikuta Kazuhiro, Kume Kiyoshi, Masamune Atsushi, Shimosegawa Tooru
Division of Gastroenterology, Tohoku University Graduate School of Medicine, Sendai, Miyagi 980-8574, Japan.
Cancer Sci. 2009 Jan;100(1):103-10. doi: 10.1111/j.1349-7006.2008.00993.x. Epub 2008 Oct 23.
We previously reported that bone morphogenetic protein (BMP)-4 induces epithelial-mesenchymal transition in a pancreatic cancer cell line. To further investigate the detailed molecular mechanism of BMP action in pancreatic cancer, we carried out comprehensive microarray analysis in Panc-1 cells. The microarray analysis elucidated novel BMP target genes, and among them, the calcium-binding protein S100P was identified as an upregulated gene. S100P induction by BMP4 was confirmed by real-time reverse transcription-polymerase chain reaction and western blot analysis in Panc-1 and HPDE cells. Short interfering RNA-based knockdown of S100P expression sufficiently repressed BMP4-induced cell migration in Panc-1 cells. Because Panc-1 and HPDE cells express wild-type Smad4, we hypothesized that Smad4 might be indispensable for S100P induction by BMP4. S100P induction by BMP4 was not observed in the Smad4-null cell line BxPC3, and was sufficiently attenuated in short interfering RNA-based Smad4-knockdown Panc-1 cells. Interestingly, detailed promoter analysis revealed that upregulation of S100P by BMP4 was independent of the Smad-binding element, indicating that an additional unknown downstream factor of the Smad4-dependent pathway is necessary for this induction. These findings are the first of their kind, and this Smad4-dependent regulation of S100P by BMP signaling might explain the migratory mechanism of cancer cells, which is still unknown.
我们之前报道过骨形态发生蛋白(BMP)-4可诱导胰腺癌细胞系发生上皮-间质转化。为了进一步研究BMP在胰腺癌中作用的详细分子机制,我们对Panc-1细胞进行了全面的微阵列分析。微阵列分析揭示了新的BMP靶基因,其中钙结合蛋白S100P被鉴定为上调基因。通过实时逆转录-聚合酶链反应以及蛋白质免疫印迹分析,在Panc-1和HPDE细胞中证实了BMP4可诱导S100P表达。基于短发夹RNA的S100P表达敲低充分抑制了Panc-1细胞中BMP4诱导的细胞迁移。由于Panc-1和HPDE细胞表达野生型Smad4,我们推测Smad4对于BMP4诱导S100P表达可能是不可或缺的。在Smad4基因缺失的细胞系BxPC3中未观察到BMP4诱导S100P表达,并且在基于短发夹RNA的Smad4敲低的Panc-1细胞中该诱导作用也被充分减弱。有趣的是,详细的启动子分析表明,BMP4对S100P的上调与Smad结合元件无关,这表明Smad4依赖途径的另一个未知下游因子对于这种诱导是必需的。这些发现尚属首次,并且BMP信号对S100P的这种Smad4依赖性调节可能解释了癌细胞的迁移机制,而该机制目前仍不清楚。