Inhibition of DACH1 activity by short hairpin RNA represses cell proliferation and tumor invasion in pancreatic cancer
Inhibition of DACH1 activity by short hairpin RNA represses cell proliferation and tumor invasion in pancreatic cancer
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DOI：
10.3892/or.2016.4843
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发表时间：
2016-08-01
期刊：
影响因子：
4.2
通讯作者：
Jiang, Zheng
中科院分区：
文献类型：
--
作者：
Bu, Xiao-Na;Qiu, Chan;Jiang, Zheng
胰腺癌是世界上最致命的疾病之一。更好地了解肿瘤发生的分子机制对提高生存率具有重要意义。人类腊肠同源物1 （DACH1）在人类恶性肿瘤进展中起着有争议的作用，其表达在多种癌症中发生改变。然而，其在胰腺癌中的功能作用和分子机制尚不清楚。通过mRNA和蛋白水平检测DACH1在胰腺癌细胞系和导管上皮细胞中的表达。设计合成了3对靶向DACH1基因的siRNA，将双链短发夹RNA （shRNA）退火后插入pGenesil-1载体中，经酶切和测序分析证实。成功构建的重组质粒被转染到Capan-1细胞中，我们的数据表明，DACH1基因表达的下调与抑制肿瘤发生有很强的相关性。CCK-8和集落形成实验表明，Capan-1细胞的增殖明显受到抑制。流式细胞术显示，干扰质粒组与对照组相比，促进了细胞凋亡（P0.05）。Transwell实验证实pshRNA-DACH1组细胞的迁移和侵袭能力明显降低。此外，我们的研究表明，DACH1表达通过与Bcl-2信号轴相互作用调节胰腺癌细胞凋亡，同时通过上皮-间质转化（epithelial-mesenchymal transition， EMT）过程控制细胞迁移和侵袭。
Cancer of the pancreas is one of the most lethal diseases worldwide. Better understanding of the molecular mechanisms involved in tumorigenesis is of great consequence to elevate the survival rate. Human Dachshund homologue 1 (DACH1) plays a controversial role in human malignancy progression with its expression being altered in a variety of cancers. Nevertheless, its functional roles and molecular mechanisms in pancreatic cancer remain unknown. The expression of DACH1 in pancreatic cancer cell lines and the ductal epithelial cells were evaluated both at mRNA and protein levels. Three pairs of siRNA targeting the DACH1 gene were designed and synthesized, double-stranded short hairpin RNA (shRNA) were annealed and inserted into pGenesil-1 vector, which was confirmed by enzymatic digestion and sequencing analyses. The successfully constructed recombinant plasmids were transfected into Capan-1 cells and our data indicated that knockdown of DACH1 gene expression showed strong correlation with repressing tumorigenesis. The proliferation of Capan-1 cells was significantly repressed as evaluated by CCK-8 and colony formation assays. Flow cymetry revealed that cell apoptosis was promoted in interference plasmid group compared with control groups (P0.05). Transwell assay validated the abilities of migration and invasion as being significantly reduced in pshRNA-DACH1 group. Furthermore, our study suggested that DACH1 expression regulates the pancreatic cancer cell apoptosis through interacting with Bcl-2 signaling axis, whereas it controls cell migration and invasion via epithelial-mesenchymal transition (EMT) process.