The present invention relates to a method for screening an anticancer agent which inhibits the binding of AIMP2-DX2 and HSP70. More particularly, the present invention relates to a method for screening an anticancer agent and an anticancer composition comprising an anticancer agent screened by the method as an active ingredient, the method comprising: a step of contacting AIMP2-DX2 or a fragment thereof with HSP70 of a fragment thereof in the presence or absence of a test substance a step of measuring the binding of AIMP2-DX2 and HSP70 a step of determining a change in the level of binding of AIMP2-DX2 and HSP70 by the test substance a step of screening a test substance which reduces the level of binding of AIMP2-DX2 and HSP70 and a step of identifying an anticancer activity of the screened test substance in cells or animals. The present invention can be effectively used to develop an anticancer agent with a completely new mechanism of action on the basis of the discovery that HSP70 is a regulator that directly binds to and stabilizes AIMP2-DX3, which is important for cancer development and progression. In addition, the pharmaceutical composition for preventing or treating cancer comprising, as an active ingredient, an anticancer agent screened according to the method of the present invention inhibits the expression of HSP70 or inhibits the binding of HSP70 and AIMP2-DX2, thereby lowering the level of AIMP2-DX2 protein, and can effectively prevent the progression of cancer. As such, the pharmaceutical composition can be useful for developing a cancer treating agent.
