Preparation Of ESF1Protein Subunit Vaccine And Nucleic Acid Vaccine Of Tirchinella Spiralis And Its Effect On Colon Cancer HCT-8

In early twentieth Century90’s,our researchers began to study the anti-tumor ofTrichinella spiralis. Although the mechanism of Trichinella spiralis anti-tumorremains uncertain, but many experiments have proved the antitumor effect ofTrichinella spiralis. In recent years there have been some reports about anti myelomacell, lung cancer, breast cancer, liver cancer effect of Trichinella spiralis. Tsp₀9347gene is screened by our laboratory in Trichinella cDNA whose expression areassociated with colon cancer HCT-8cells. Tsp₀9347gene encodes the ESF1protein.our laboratory found that ESF1protein antibody has an inhibitory effect on humancolon cancer cell line HCT-8.On this basis, this study intends to make ESF1protein subunit vaccine andnucleic acid vaccine of Trichinella spiralis.To observe the effect of ESF1proteinsubunit vaccine and nucleic acid vaccine on human colon cancer HCT-8cells. Usingimmunohistochemistry to observe the changes of mouse spleen CD8, CD4,CD20. Inorder to provide the theoretical support of Trichinella spiralis anti colon cancer, andexplore new ideas for the development of colon cancer vaccine.This experiment using cDNA preserved by the the laboratory of parasites andparasitic diseases of animal medicine, Jilin University School as a template. Get theTsp₀9347gene by PCR. Verified the target gene by agarose gel electrophoresis,thenrecombine plasmid pMD-18T-Tsp₀9347.After double enzyme digestion to verify thetarget gene, connected Tsp₀9347gene to pcDNA3.1（-） and pET-28a（+）.Then thepET-28a（+）-Tsp₀9347and pcDNA3.1（-）-Tsp₀9347were digested to verify. Theprokaryotic expression plasmid pET-28a （+）-Tsp₀9347was transformed into E. coliBL21, induced expression and purification, and through the SDS-PAGE and WesternBlotting to verify the expression of protein; The eukaryotic expression plasmid pcDNA3.1（-）-Tsp₀9347was transformed into Escherichia coli DH5alpha andextracted, then transfected into Hela cells, and through SDS-PAGE and WesternBlotting to verify the expression of protein. A large number of purified prokaryoticexpression protein and eukaryotic expression plasmid were mixed with thecorresponding adjuvant to make the ESF1protein subunit vaccine and nucleic acidvaccine of Trichinella spiralis The mice were transplanted under the kidney capsule oftumor cell model （SRCA） to observe the effects of two kinds of tumor vaccine oncolon cancer cell line HCT-8. HE staining showed that after treatment, the vaccinesignificantly reduced the number of tumor cells, the fibroblasts proliferated around thetumor cells, inflammatory cell infiltration in some regions. Subsequently we usedimmunohistochemical method to observe the change of CD8, CD4, CD20in micespleen, results show, mice spleen CD8, CD4, CD20expression in the vaccine groupwas significantly higher than the control group. Conclusion: the success of preparationof subunit vaccine and nucleic acid vaccine of Trichinella spiralis ESF1protein, andconfirmed that has certain inhibiting effect on colorectal carcinoma HCT-8cell line.The mechanism may be related to the immune response of cells and humoral in mice.