The Impact Of The CIK Cells On Drug-resistant Human Lung Adenocarcinoma A549/DDP Cells

Objective:To investigate the effects of CIK cells on cisplatin resistant human lung adenoearcinoma A549/DDP cells.Methods:PMBC was extracted from the human whole blood using ficoll gradient centrifugation. IFN-1, IL-1, IL-2and anti-CD3monoclonal antibody were added so as to induce PMBC to transit into CIK cells in vitro. Using Leica microscope to observe the morphological features of A549/DDP cells and CIK cells. Then CIK cells were co-cultured with A549/DDP cells. According to different target ratio of CIK cells to A549/DDP cells, the groups were divided into four groups including40:1target ratio group,20:1group,10:1group and no CIK effect group as the control group. The growths of A549/DDP cells were also investigated under Leica microscope. Concerning gene and protein expression, experiments were divided into five groups:A549cells, A549/DDP cell, A549/DDP cells in IL2-culture medium, CIK co-culture with A549/DDP (10:1) and CIK group. Using PCR, RQ-PCR and Western blot to observe the different experimental groups gene (mdr-1, of mrp, gst-π) transcription and protein (P-gp, MRP, and GST-π) expression.RESULTS:CIK cells which equably suspended in the medium were round or oval; while A549/DDP cells which grew adherent to the surface were polygon, mulberry-like. When CIK cells and A549/DDP cells were co-cultured, CIK cells tended to play as a killer to A549/DDP cells. In the10:1target ratio group, the growth of A549/DDP cells were inhibited. When the target ratio increased to40:1, there was almost no survival A549/DDP cells. RT-PCR and RQ-PCR detected that the Mdr-1, mrp, gst genes are involved in A549/DDP cells resistant to cisplatin and CIK cells inhibited A549/DDP cells resistance mdr-1, mrp and gst-π gene transcription; Western blot analysis showed that the P-gp, mrp, proteins are involved in A549/DDP cells resistant to cisplatin and CIK cells down-regulated A549/DDP cells resistance proteins including P-gp (P-glycoprotein), MRP and GST-π protein expression.CONCLUSION:CIK cells obviously killed cisplatin-resistant human lung adenocarcinoma A549/DDP cells. Mdr-1, mrp, gst genes and proteins are involved in A549/DDP cells resistant to cisplatin.CIK cells down-regulated some important A549/DDP cells resistance genes and proteins expression which may inferred that CIK cells can reverse A549/DDP cells resistance to cisplatin.