The Anti-tumor Efefcts Of Trichinella Spiralis Protein Anitbody Which Associated With MCF-7Breast Cancer Cell

In this study, a correlation between antigen protein in Trichinella spiralis andMCF-7breast cancer cells determined by ELISA and Western blotting method.TS498gene protein which related to MCF-7cells was obtained by immunoscreeningfrom Trichinella spiralis cDNA expression library, and then observed its anti-tumoreffects by animal studies.Preparation of associated antigen: First preparation of anti-Trichinella spiralislarvae soluble protein hyperimmune sera, and anti-MCF-7breast cancer cell antigensera, respectively. And then the cross-reactivity between Trichinella spiralis antigenand MCF-7antisera or between MCF-7antigen and Trichinella spiralis antisera weremeasured by ELISA. ELISA results preliminary showed that there are cross-reactionbetween Trichinella spiralis and MCF-7cells, existed associated antigen. Then theimmune response between Trichinella spiralis antisera and MCF-7breast cancer cellantigen was detected by western-blotting. Five specific reaction bands wereobtained.compare with the standard molecular weight protein Marker, the reactionantigen presented near18kDa,29kDa,32kDa,56kDa,60kDa. Then thecorresponding reaction antigens were separted by polypeptide protein separationcolumn (ultrafiltration) from the MCF-7antigen. And then they were used to preparethe antibodys.Screening MCF-7tumor-related antigen protein gene from Trichinella spiralis:Associated antigen antisera as a screening antibody, immunoscreening Trichinellaspiralis cDNA expression library for positive plaques, rescreening positive plaquesuntil obtain the consistent plaques. Then the phagethe plaques DNA were extracted,the inserted DNA fragments were amplified, and the protein gene were sequencingand bioinformatics analyzing. The results are access to an open reading frame size of498bp, which was small heat shock protein belonging to the Trichinella spiralis heat shock protein family. The gene was coloned and expressed in E. coli, the the purifiedTS498fusion protein was use to immunize the mice for the antisera. Vitroimmunofluorescence test shows that the TS498protein antisera could conjunctionwith the MCF-7cells.Observation of the anti-tumor effect on the nude mice which were treated by theTS498antisera: The nude mice with breast cancer solid tumor model wereeatablished by inoculated with the fresh alive MCF-7cells,20d later, thetumor-bearing nude mice were treated with sera for one month, during the treatment,the mental state of the mice, the tumor growth condition, the changes of the tumorsize and weight, and the mortality rate of the mice were observed and recorded. Onemonth after treatment, mice were sacrificed and the tumor weight, volume, and theinhibitory rate were calculated. The results show that the antiserum of MCF-7associated antigen TS498proteins has anti-tumor effect, the highest inhibition rate upto36.12%, and its therapeutic effect was dose-dependent. These results provided theavidence for researching Trichinella spiralis anti-tumor effect from different aspects.Also provides a reliable scientific basis for the R&D of anti-tumor biological agents.