Expression Of SFRP5 In Pancreatic Cancer And The Construction Of Lentivitial Vector SFRP5-shRNA

Objective: To investigate the expression and clinical significance of SFRP5 gene in pancreatic cancer, to construct SFRP5-shRNA lentivirus vector, to provide experimental materials for further study of SFRP5 in pancreatic cancer cell lines and its mechanism.Methods: To collect pancreatic cancer paraffin from 2010 to 2013 in general surgery of the first hospital of Lanzhou University. To choose 71 specimens with a clear pathological diagnosis and clinical follow-up data,use immunohistochemistry to detect the expression of SFRP5 in pancreatic cancer, and analyze the relationship between SFRP5 expression,clinicopathological factors and prognosis; Using West-bloting and qRT-PCR to extract protein and mRNA,to detect the expression of SFRP5 in fresh pancreatic cancer. According to people SFRP5 genetic information published in GenBank, to build four SFRP5-shRNA lentivirus interfering vector,using 293 T cells to carry out lentiviral packaging and to determin the virus titer.Results: The expression of SFRP5 in pancreatic cancer tissues was significantly lower than the adjacent tissues(40.8% vs 85.7%, P <0.05), SFRP5 protein and pancreatic cancer TNM stage(P <0.05), distant metastasis(P <0.05), peripancreatic violations(P <0.05) and prognosis are relevant, and was an independent risk factors for pancreatic cancer prognosis. After successfully constructing SFRP5-shRNA lentivirus vector transfected 293 T cells, green fluorescence can be seen under a fluorescent microscope, and viral packaging, slow purified virus particles, measured virus titers 1 * 109 TU / ml.Conclusion: The expression of SFRP5 was disappeared in pancreatic cancer, it was an independent factors affecting the prognosis of patients with pancreatic cancer, SFRP5 low expression, presence of distant metastasis, TNM stage and late patient peripancreatic invasion existed a poor prognosis in pancreatic cancer. Through RNA interference technology, successfully constructing SFRP5-shRNA lentivirus vector to obtain a higher viral titer after packaging, the infection of pancreatic cancer cell lines for subsequent cell biology studies of pancreatic cancer and provide experimental basis.