| Background:Pancreatic ductal cell adenocarcinoma (PDAC) is one of gastrointestinal carcinomas with the poorest prognosis and diagnostic difficulty which poses large threat to people’s health and life. The 5-year survival rate is reported to be 6.8% to 25%, and the median survival time of postoperative patients is ranging from 8 to 12 months. We had detected more than 80% of PDACs at an advanced stage due to its specialty in biological behavior and anatomical position, and missed the opportunity to undergo radical surgery. The only way to improve patient’s prognosis is making an accurate diagnosis in its early stage. In addition, lymph node metastasis is an independent risk factor which affects the survival of postoperatives. Patients with lymph node metastasis have apparently shorter survival times, most of them subsist no more than 2 years. It is imperative to find some tumor markers with comparatively high sensitivity and selectivity. Heparanase (HPSE) is an endoglycosidase which exclusively degrades heparan sulfate (HS) side chains. HS is a kind of mucopolysaccharides which is of abundant presentation on cell surface and in extracellular matrix. HPSE is actively involved in the remodeling of subendothelial and subepithelial basement membrane, as well as neoplasm metastasis and angiogenesis. Lymphatic endothelial cells possess high level of VEGFR-3 which has the potential of binding vascular endothelial growth factor C (VEGF-C), as a result, initiating lymphangiogenesis and heightening lymphatic vessel density (LVD).It may inevitably lead to the invasion into lymphatic system. HPSE and VEGF-C have long been considered as the key cytokines which promote metastasis and angiogenesis in head and neck neoplasms. However, the relationship between them in PDAC is still unclear.Objectives:To build the recombinant human HPSE plasmid and the small interfering RNA targeting HPSE gene. To investigate its correlations with VEGF-C expression and metastatic capability of pancreatic cancer cells in vitro. To figure out how the suppression of target gene may affect apoptosis of transfected BxPC-3.Methods:BxPC-3 was transiently transfected with GV230/HPSE and HPSE-siRNA. The expression levels of HPSE and VEGF-C were detected by performing florescent real-time quantitative PCR (RT-qPCR) and Immunoblotting. The metastatic potential of treated BxPC-3 was evaluated by Transwell invasion assay and Srcatch-wound assay. Then we assessed cell apoptosis by flow cytometry.Results:By RT-qPCR, there was a 10.7-and 3.24-fold (All P<0.01) elevation of HPSE mRNA and VEGF-C mRNA in GV230/HPSE group, respectively, whereas siRNA group showed the opposite result (-2.45-fold, P<0.01;-1.84-fold, P<0.01). The same tendency was detected in immunoblot assay. In invasion chamber assay,138+5 cells in GV230/HPSE group and 53±4 cells in siRNA group invaded through the Matrigel, which was significantly different from the control group. The same trend was found in Srcatch-wound assay. Flow cytometry analysis implicated that interference of HPSE increased the number of apoptotic cells slightly (8.65%)Conclusions:HPSE regulates the expression of VEGF-C and facilitates metastasis of BxPC-3 in vitro. Suppression of HPSE enhances apoptosis. |
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