Treatment Of Mice Breast Cancer With Dendritic Cells Loaded With Heat-treated Peptide Of Tumor Cells

Research backgroundbreast cancer is the most popular maglignant tumor among women,and it's disease incidence rate is becoming more and more higher.At present,the he major strategy for treatment of breast cancer includes surgery, chemotherapy, radiotherapy and endocrinotherapy,enven though, a radical cure effects is still impossible, the high recurrence rate and poor prognosis is hard to be changed.Due to this, biotherapy is increasingly recognized by more and more people,and the characterization of tumor antigens recognized by immune effector cells has opened the perspective of developing therapeutic vaccines in the field of maglignant tumors.The tumor vaccine, due to it's potential advantages including:the induction of a robust immune response against tumors that are spontaneously weekly immunogenic, the long-term immune memory, critical to prevent efficiently tumor recurrence and so on,is more and more widely used to treat various tumors like beast cancer,renal cancer, melanoma and has been proved to be efficient. Because of this,the vaccine based on dentritic cell has becomes a new focus research in the biotherapy of breast cancer; The way of making dendritic cell-based vaccines with was widely accepted duo to the cell peptide could be prepared and it's good therapeutic effect.The dendritic cell-based vaccines were used to treat the malignant tumors such as non-Hodgkin lymphoma,myeloma, melanoma, non-small-cell lung carcinoma,prostatic cancer, colorectal cancer,and received invariably therapeutic effect in clinical trials.But,most of the tumors'antige ofen have weak immunogenicity,this restrict the capability of dendritic cells to present the tumor antige and active the specific Cytotoxic T lymphocyte and restrain the improve of the DCs-based vaccines'curative effect.For this,the human think a way to achieve the target by adding the immune adjuvant like heat shock protein, bacillus aeruginosus to tumor's antige and it works in vitvo. The heat shock protein is called molecular chaperones,it ia a kind of protein which can protect the cell when meet an emergency, it work in the progress of certain protein's foldback, assemblage, intracellular transportion and protein degradation.lt can change the activity and function of some protein, besides, it can work as immune adjuvant and can active the dendritic cells's function.In this research, we make two different DCs-based vaccines by mice breast cancer antige which were treated by heat-treated and freeze-thrawing way, respectively.then, the two different DCs-based vaccines' therapeutic effects were compared.Research objects1. To research the way of culturing mouse marrow dendritic cells with Granulocyte-Macrophage Clone stimulating factor, Interleukin-4 and Tumor necrosis factor-a.2. To research the expression of Heat Shock Proteins-70 in the EMT6 breast cancer cell made by heat-treated and freeze-thrawing ways, respectively.3. The acquired tumour antigen was co-incubated with dendritic cells that had been cultivated in vitro so as to prepare dendritic cells tumor vaccine.And then the different dendritic cells tumor vaccines were used to treat mice breast cancer model and compare the therapeutic effects of two different dendritic cells tumor vaccine.4. Kill the mouses at 21 day, measure and compare the weight and volume of tumor and the concentration of INF-γ,IL-2 and IL-12(P70) in blood-serum of mouses in different group.Research methods1.Observe the appearance of dendritic cells regularly, detect the cells'phaenotype use flow cytometry.2.Detect the content of heat shock proteins 70 in tumor cells treated by different ways.3.Set mice breast cancer model and group random, then treat different group with different dendritic cell vaccines.4.At the 21-th day,kill the mouses, measure and compare the weight and volume of tumor, the concentration of INF-γ,IL-2 and IL-12(P70) in blood-serum of mouses in different group was measured by enzyme linked immunosorbent assay(ELISA). Research results1.It was proved by observation under light microscope and flow cytometry that the cells induced by Granulocyte-Macrophage Clone stimulating factor, Interleukin-4 and Tumor necrosis factor-a were typical mature dendritic cells.2.The content of heat shock proteins 70 in the heat-treating tumor cells is much more higher than those in the freeze-thrawing tumor cells.3. Compared with freeze-thrawing DC tumor vaccine group and the contol group,the tumor size and weight of heat-treated DC tumor vaccine group were significantly small/light (p<0.05) 4.Compared with freeze-thrawing DC tumor vaccine group and the contol group, the concentration of INF-γ,IL-2 and IL-12(P70) in blood-serum of heat-treated DC tumor vaccine group were high significantly (p<0.05)ConclusionsDue to the heat shock proteins works as immune adjuvant,compared with the DC tumor vaccine made by freeze-thrawing tumor peptile, the DC vaccine tumor made by heat-treated tumor peptile is more efficient for mice breast cancer through eliciting more intensive antitumor immunity effect, and it can be an effective adjuvant to the routine therapy of breast cancers.