The invention relates to a preparing method of lecithin of tuna roes. The method comprises the steps of 1) enzymolysis,wherein distilled water and trypsin are added into tuna roe homogenate for enzymolysis, and centrifugation is conducted after inactivation to remove supernatant; 2) digestion, wherein acetone is added into a filter cake after enzymolysis, stirring and suction filtration are conducted, and then acetone is utilized for digestion until the color of the acetone does not change; 3) secondary digestion, wherein absolute ethyl alcohol is added into the filter cake after digestion, and microwave digestion is conducted intermittently; 4) purification, a CHCl3-CH3OH(2:1) mixed solution is taken as an eluent to conduct silica gel chromatography column purification on a crude lecithinproduct, and the eluent is collected and dried to obtain the pure lecithin product. By means of the method, the extraction speed of free phospholipid can be increased, the extraction rate is increased, and then the finished product lecithin can be obtained through silica gel chromatography column purification; the purity is improved on the basis of significantly increasing the extraction rate ofthe lecithin, further direct application of the lecithin is facilitated, and the lecithin of the tuna roes can be applied to inhibition of human liver cancer HepG2 cells and lipid decreasing of Changliver cells.