The invention relates to a CAR-NK cell as well as a preparation method and an application thereof. The method comprises the following steps: S1, linking a specific antibody coding gene into a cloning site region of a modified CAR skeleton so as to obtain a CAR gene, wherein the modified CAR skeleton comprises a CD244 episporium signal region, the cloning site region, a CD244 extracellular hinge region, a CD244 transmembrane part region and a CD244 intracellular part region which are sequentially connected; and S2, infecting the CAR gene with NK cell through lentivirus, so that the NK cell expressing the CAR protein is obtained, namely the CAR-NK cell. The prepared tumor immnuotherapy cell, namely the CAR-NK cell, of the invention, besides a patient with cancer, is also applicable to allosome infusion; therefore, the prepared tumor immnuotherapy cell, in comparison with existing CAR-T, is wider in application scope and is lower in preparation cost.