Cloning, expression and characterization of a HER2-HER3 DNA as a novel immunotherapy agent

Introduction: Breast cancer(BC) is one of the common disease among women worldwide and about 2.1 million new cases were diagnosed in 2018.Despite of effective treatments such as chemotherapy, radiotherapy and Hormone therapy BC has been remained one of the significant reasons of women ‘s death. Recent progress in the new field called immunotherapy has changed the management of breast cancer completely and plays a major role in the efficacy improvement of BC conventional treatments. One of the significant type of Antigen-specific immunotherapy is DNA vaccine. This type of vaccine is the result of designing an expression vector which encodes a special antigen and in the most cases this antigen is over expressed in that specific type of cancer. Cancer vaccines have led to modest effects in treatment of cancer affected patients due to immunosuppression in micro environment of Tumors. cytokine-based adjuvants such as interleukin (IL)-12 have been suggested as one of the most effective enhancing immune responses optionsMaterials & Methods: The nucleotide sequences of HER2 and HER3 and CCL20 were selected according to relevant studies. The selected sequences submitted to the Basic Local Alignment Search Tool (BLAST) to check the conservation of the selected Sequences. These sequences were linked together using a linker. The GFP tag was designed at the N terminal of the construction. ,The CMV promoter was designed and the selected sequences were designed between two restriction enzyme sitesIn this study Bioinformatics Tools and In-silico prediction methods were used in order to select the appropriate HER2, HER3, CCL20 sequences and the standard form of the recombinant Vector(PIRES2-GFP). PIRES2-GFP transfection in to HECK 293 T cell line was carried out.Results: According to previous studies and bioinformatics analysis, we selected three main sequence of HER2,3 and ccl20.24h after transfection of recombinant srtructure,green light emission was detected under fluorescence microscopeConclusion & discussion: The study results indicated that the recombinant structure has the potential to be expressed and this structure has the capacity to use in animal models in order to evaluate the immunogenicity.DNA vaccine constructions have indicated promising results and they can use seperatyely or in combination with other conventional treatments in order to improve the efficacy of the treatment.

کلیدواژه ها:

Breast cancer ، DNA vaccine ، HER2 ، HER3