The invention discloses a novel screening method of anticancer medicaments. The novel screening method is characterized by comprising the followinh steps of: (1) preparing TTLL12 overexpressed Hep-2 cell lines; (2) selecting positive control and negative control medicaments; (3) obtaining the optimum concentration of nitrotyrosine and the optimum diluted concentrations of medicaments to be detected; and (4) screening the medicaments, finally checking readings and comparing. The novel screening method of the anticancer medicaments disclosed by the invention has the characteristics that the method is simple, a user only needs to master a cell culture and Elisa (Enzyme-linked Immuno Sorbent Assay) technology and then masters the law, the method is fast and cheap, the method is few in reagents and variation factors, stable, high in sensitivity, good in specificity, easy to popularize and capable of screening in a large scale. Furthermore, through the experiment, whether the medicaments to be detected have anti-cancer effects and anticancer effect mechanisms of the medicaments to be detected simultaneously can be obtained, so that the speed of clinically applying the medicaments to be detected is greatly accelerated, and more lives of patients are expected to be saved.