Cancer metastasis is one of the most serious problems for tumor therapy,which is closely related to cell adhesion and deadhesion process.Better comprehension of cell adhesion ability will benefit drug research.Here,a biomimetic microfluidic enzyme digestion method was proposed to gently measure the influence of drugs on cell-matrix adhesion ability at the single cell level.The method can selectively digest the extracellular matrix (ECM) that linked to a single cell,and the trypsin concentration around the cell is relatively uniform and constant,thus the measured cell adhesion strength should be precise.Commercially available anti-cancer agents including 5-fluorouracil (5-FU),actinomycin D (Act D),temozolomide (TMZ) and allicin were evaluated,and the data showed only TMZ and allicin can inhibit cell adhesion significantly under our experiment conditions.The influence of TMZ became more and more obvious as the increase of duration and the effect became prominent only after 6 h adhesion process,which could provide a quick evaluation of whether the drugs are effective to cancer cell (compared with Calcein-AM/PI cell viability test).The adhesion strength of U87 cells decreased when the concentration of TMZ increased,and the effect of TMZ can be effectively inhibited by adding lactic acid to culture medium,which indicated acidic tumor microenvironment could promote drug resistance of tumor cells.Different from conventional evaluation methods which focus on the drugs' influence on cellular viability or metabolism,this work provides a new perspective to study the effect of drugs,which is helpful to enrich the drug evaluation system.