Experimental Study Of Anti-HER-2/neu Monoclonal Antibody (Herceptin) Combined With CIK Cells On Breast Cancer Cells In Vitro

Objectives:1. To observe the proliferation of CIK cells and their killing tumor ability on breast cancer cell in vitro,and to compare the effects of different opponents in the cell culture systems so as to determine whether CIK cells are more efficient than LAK cells or not.2. To investigate the influence of HER-2/neu monoclonal antibody (Herceptin) on the proliferation of breast cancer cells excessively expressing HER-2/neu in vitro,and to evaluate the latent application value of the immunotherapy of breast cancer.3. To evaluate the synergistic effect of Herceptin combined with CIK cells so as to exploit a new method of the immuno therapy of breast cancer.Methods:1. Inducing and amplifying the immune effector cells in vitro with the cell culture plus cytokine and assaying their phenotypes with FCM.2. Detecting the cytotoxicity of Herceptin combined with or not with CIK cells,different effector cells,and dfferent components in the culture systems in vitro with MTT respectively.3. Demonstrating the synergistic effect of Herceptin plus CIK cells on breast cancer cells compared with their corresponding control group by the following methods:low concentration Herceptin plus moderate-effective CIK cells( no Herceptin in the control group);moderate concentration Herceptin plus low-effective CIK cells( no CIK cells in the control group).Results:1. The CIK cells expressing both CD3,and CD56 amplified largely while the LAK cells seemed no remarkable amplification when the cells were cultured,. The cytotoxicity of the CIK cells in vitro was higher than that ofLAK cells. In the CIK cells culture systems,the killing tumor cell activities of CIK cells plus superant,CIK alone and superant alone were 95.78+5.16%,55.42+3.10%,27.65+2.08%,respectively.2.Herceptin expressed significant inhibition on SKBR3 and MNA-MB-453 cells. The rates of inhibition on AKBR3 and MNA-MB-453 at 5 ig/ml was 26.19%,13.00% and at 80ug/ml 50.12%,32%,respectively.3. The anti-breast cancer cell effect of Herceptin combined with CIK cells was significantly superior to that of Herceptin or CIK cells alone. After culturing the tumor cells 92 hours with Herceptin at 2.5ng/ml,and then 4 hours with CIK cells(E:T=10:l),the rate was 32% higher than CIK alone,while that of Herceptin 80ug/ml plus CIK cells(E:T=50:l) was 18% higher than Herceptin alone.Conclusion:1.CIK cells are new kinds of immunocytes which have excellent proliferation ability. The components of superant have some killing tumor activity and can enhance the effect of CIK.2. Herceptin can remarkably inhibit the proliferation of the breast cancer cells highly expressing HER2/neu receptors and have the synergistc effect with CIK,which manifests latent application values of the immunotherapy of breast cancer.