Method for screening anticancer substances, set or kit for implementing said method

Abstract

A method for screening anticancer substances including the following steps: a) obtaining a culture of tumour cells which do not express E-cadherin on their cell membrane; b) bringing the cells obtained at step a) into contact with a candidate substance or a combination of candidate substances; c) detecting, in the culture of cells obtained at the end of step b), the presence of E-cadherin at the cell surface; d) positively selecting the candidate substance or combination of candidate substances when the E-cadherin has been detected at step c).

1 . A method for screening antimetastatic anticancer substances which do not act directly on the expression of E-cadherin, said method comprising the following steps:
 a) growing tumour cells with the capacity for forming metastases and which do not express E-cadherin on their cell membrane;   b) bringing the cultivated cells obtained at step a) into contact with a candidate substance or a combination of candidate substances;   c) detecting, in the culture of cells obtained at the end of step b), the presence of E-cadherin at the cell surface;   d) positively selecting the candidate substance, or the combination of candidate substances, when the E-cadherin has been detected at step c).   
     
     
         2 . The method according to  claim 1 , wherein at step c), the detection is carried out by bringing the cells obtained at the end of step b) into contact with a ligand compound of E-cadherin. 
     
     
         3 . The method according to  claim 2 , wherein the E-cadherin ligand compound binds to the extracellular portion of E-cadherin. 
     
     
         4 . The method according to  claim 2 , wherein the ligand compound is labelled with a detectable molecule. 
     
     
         5 . The method according to  claim 2 , wherein at step c), a secondary ligand compound that binds to the E-cadherin ligand compound is used, the said second ligand compound being labelled with a detectable molecule. 
     
     
         6 . The method according to  claim 2 , wherein the ligand compound consists of an anti-E-cadherin antibody. 
     
     
         7 . The method according to  claim 6 , wherein at step c), a secondary ligand compound that binds to the anti-E-cadherin antibody is used, the said second ligand compound being labelled with a detectable molecule. 
     
     
         8 . The method according to  claim 7 , wherein the secondary ligand compound comprises a streptavidin molecule coupled to, or fused with, an enzyme. 
     
     
         9 . The method according to  claim 7 , wherein the second ligand compound is an antibody directed against the anti-E-cadherin antibody. 
     
     
         10 . A set or kit for screening antimetastatic anticancer substances which are capable of reversing metastatic phenotype to a non metastatic phenotype and which do not act directly on the expression of E-cadherin, said set or kit, comprising:
 a) tumour cells having the capacity for forming metastases and which do not express E-cadherin on their cell membrane; and   b) a ligand compound of E-cadherin.   
     
     
         11 . The set or kit according to  claim 10 , wherein the E-cadherin ligand compound binds to the extracellular portion of E-cadherin. 
     
     
         12 . The set or kit according to  claim 10 , wherein the E-cadherin ligand compound consists of an anti-E-cadherin antibody. 
     
     
         13 . The set or kit according to  claim 10 , wherein it also contains a secondary ligand compound capable of binding to the E-cadherin ligand compound, the said second ligand compound being labelled with a detectable molecule. 
     
     
         14 . The set or kit according to  claim 13 , wherein the second ligand compound is an antibody directed against the anti-E-cadherin antibody. 
     
     
         15 . The set or kit according to  claim 14 , wherein the secondary ligand compound comprises (i) an antibody directed against the anti-E cadherin antibody and (ii) an anti-peroxidase antibody, said anti-peroxidase antibody being bound to two peroxidase molecules at its antigen binding site (CDR).