Studies On Inducing Apoptosis Effects And Mechanism Of CIK Cells On Zk-75-1 Breast Cancer Cell Lines

Objective: To study the anti-proliferation and inducing apoptosis effects of cytokine-induced killer cells-CIK cells for breast cancer cell lines and to investigate its underlying mechanism.Methods: To detect the anti-proliferation and the cytotoxicity Of CIK cells on breast cancer cell line by MTT assay.The morphological changes of the apoptosis cell were observed by invert microscope HE stain and transmission electron microscope . Inducing apoptosis of CIK cells on breast cancer cell line was evalrated by TUNEL methods-, The positive expression of p53 p16 C-myc Bcl-2 and bax were determined by immunocytochemistry(ICC).The expression changes of relating-apoptosis gene proteins were studied.Results: (1) MTT assay showed that the inhibitive rate inhanced obviously with theaddition of Effect/Target rate and extension of time .There are significantly differences between different E/T rates at the same time (P<0.01).There are significantly differences at different time and the same E/T rate(P<0.01); (2) Invert microscope observation showed that CIK cells removed to the target cells and revoked round cancer cells and appeared typical change of rose circulatity.Granule substance appeared in tumor cells cytoplasm.Some CIK cells removed into the tumor cells cytoplasm or nucleolus.Only granule shape patch were restained in some tumor cells .Breast cancer cells lived well in control group; (3) HE stain observation showed that the CIK group was the same with in invert microscope; (4) Transmission electron microscope observation showed that chromatin condensed and nucleolus disintegrated in many target cells and vacuoles were found at cytoplasm and apoptotic boby formed after effect cells and target cells cultivated together four and twelve hours.Many targets cellsdied by apoptosis and necrosis after effect cells and target cells cultivated together twenty-four hours; (5) TUNEL showed that the cells of control group were not to be stained or be stained average light blue and the cells of experiment group became small and mucleolus were dyed deeply blue. The apoptosis rate of CIK group increased in 4-12hours and declined in 12-24 hours.The apoptotis rate of CIK groups have significantly differences compared with control group(P<0.01) ; (6) Immunocytochemistry showed that expression of p53, p16 C-myc Bcl-2 protein in CIK group inclined and Bax protein increased when time was continued and there were different significantly compared with control (P<0.01).Conclusions : 1.There are anti-proliferation and inducing apoptosis effects for CIKcells against ZK-75-1 breast cancer cells.2.CIK cells induce apoptosis of ZK-75-1 breast cancer cell line before twelve hoursand induce necrocytosis after twelve hours.One of the mechanisms may be inhanceBax proteins expression and decline p53 p16 Bcl-2 and C-myc proteins expression oftumor cells.3.CIK cells are highly efficient cytolytic effector cells which have a strongersignificant suppression against growth of breast cancer and the result provides anexperimental basis for CIK to clinical application as a adoptive immunotherapy.