The Expression And Clinical Significance Of Human Tumor-related Protein HyrdC In Gastric Cancer

Objective To prepare pure hyrdC prokaryotic expression protein and the monoclonal antibody againt hyrdC, and to detect the hyrdC protein in gastric cancer with this monoclonal antibody. Methods The target DNA sequence of hyrdC was obtained by RT-PCR and sequently inserted into prokaryotic expression vector pET32a. The TRX-hyrdC fusion protein was purified.The 6-week-old BALB/c female mice were immunized with recombinant fusion protein TRX-hyrdC to prepare monoclonal antibodies with routine method. The hybridoma cells were primarily screened by enzyme-linked immunoabsorbent assay(EIISA), and identified by Western blot. With previous prepared monoclonal antibody, We detected the expression of hyrdC protein of gastric cancer and normal tissues of adjacent tumor by immunohistochemical method (SP) Results The sequence of hyrdC ORF was conformed to the report in GenBank. The monoclonal antibody against hyrdC has been successfully obtained, These monoclonal antibodies were demonstrated to combine with human hyrdC protein specifically by Western blot. SP results showed that the expression of hyrdC protein in gastric cancer is much more significant than that in normal gastric mucous membrane, (P<0.001)o Conclusion there were significant differences between the gastric tumor tissues and normal tissues...