Inhibition of DACH1 activity by short hairpin RNA represses cell proliferation and tumor invasion in pancreatic cancer

Abstract

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 (P<0.05), whereas cell cycle had no significant differences among the groups (P>0.05). 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.

DACH1 suppresses breast cancer as a negative regulator of CD44

“…However, its absent or lower expression contributes to the initiation and progression of various tumor types, including lung adenocarcinoma 14 , pancreatic cancer 47 , breast cancer 10 and gastric cancer 48 . This study provided results that expression types of DACH1 and CD44 were opposite in breast cancer.…”

“…DACH1 played important roles in invasion and migration of various neoplasms, such as lung adenocarcinoma 14 , gastric cancer 48 , pancreatic cancer 47 and breast cancer 12 . Several lines of evidence showed that DACH1 is absent or suppressed in poor prognosis breast cancer 58 .…”

FOXM1-induced miR-552 expression contributes to pancreatic cancer progression by targeting multiple tumor suppressor genes

“…In our study, we identified three tumor suppressor genes, DACH1, PCDH10 and SMAD4 as targets of miR-552 in pancreatic cancer. DACH1 is a paralog of Drosophila dachshund (dac) gene in vertebra, which is expressed widely in normal adult tissues, and loss of DACH1 expression predicts poor outcome in many cancers [33]. PCDH10 belongs to the non-clustered protocadherin protein family encoding calciumdependent adhesion protein and is known as a tumor suppressor gene in many tumors.…”

Transcriptome and Network Dissection of Microsatellite Stable and Highly Instable Colorectal Cancer

“…Previous studies reported methylation of DACH1 promoter and its downregulation in cancer and demonstrated that DACH1 can inhibit breast tumor invasion and growth by suppressing epithelial-mesenchymal transition (EMT) and proliferation through repression of Snai1and Cyclin D1, respectively (Wu et al, 2006;Zhao et al, 2015). Bu, Xiao-Na and colleagues illustrated that DACH1 upregulation can promote pancreatic cancer growth and invasion; downregulation of DACH1 activity with shRNA can repress cell proliferation and tumor invasion by mostly inducing apoptosis and inhibiting EMT in pancreatic cancer cells through modulating Bcl-2 (pro-survival regulator) and E-cadherin, respectively (Bu et al, 2016). Our data have shown significant upregulation of DACH1 in MSS CRC; moreover Cyclin D1 and Snai1 represent upregulation in 3 out of 4 MSS studies; on the other hand, Bcl-2 was downregulated and E-cadherin did not represent significant change in expression in MSS studies.…”