The invention provides a method for effectively amplifying cytokine-induced killer (CIK) cells and improving specific tumor killing capability of the CIK cells. The method comprises the following steps: separating to obtain mononuclear cells; transferring the obtained mononuclear cells to a culture flask; adding CD3mAb with the final concentration of 400-600 ng/ml, CD28mAb with the final concentration of 1400-1600 ng/ml, IFN-gamma with the final concentration of 4000-4800 IU/ml and IL-2 with the final concentration of 4000-4800 IU/ml, IL-15 with the final concentration of 140-160 ng/ml; on the fourth day, replenishing fresh culture solution including 600-1000 IU/ml of IL-2 and 20-30 ng/ml of IL-15; on the sixth day, culturing the cells in separated flasks; on the eighth and ninth days, respectively transferring cells of the flask to a cell culture bag of 1.8 L, and replenishing fresh culture solution including 600-1000 IU/ml of IL-2 and 20-30 ng/ml of IL-15; at the eleventh day, culturing the cells in separated bags, and replenishing fresh culture solution including 600-1000 IU/ml of IL-2 and 20-30 ng/ml IL-15; continuously inducing and amplifying till the cells are obtained from the fifteenth day to the twenty-first day; incubating the obtained cells for 30-40 minutes at the room temperature by utilizing anti-EGFR and anti-CD3 bifunctional antibodies, washing the incubated cells in normal saline, and collecting the cells, thereby obtaining specific tumor-killing CIK cell preparations.