Method and kit for the prognostication of breast cancer

Abstract

This invention relates to a method, kit and controls for detecting HER-2/neu gene amplification as a predictor of breast cancer reoccurrence and patient survival. The method is a fluorescent in-situ hybridization (FISH) assay using a labeled DNA probe. By determining the genetic nature of the cancer cells, appropriate treatment may be utilized. Control tumor cell lines with predefined amounts of HER-2/neu gene amplification are also disclosed.

We claim:  
     
         1 . A method for determining the likelihood of cancer recurrence in a patient comprising the steps of: 
 (a) obtaining tumor cells from the patient; (b) applying to each of said cells a detectable probe capable of hybridizing to the HER-2/neu gene;    (c) counting the number of HER-2/neu genes per cell; and    (d) determining the likelihood of cancer recurrence wherein an average of about 10 or more HER-2/neu genes per tumor cell indicates a high likelihood of cancer recurrence and an average of about 3 or fewer BER-2/neu genes per tumor cell indicates a low likelihood of cancer recurrence.    
     
     
         2 . The method according to  claim 1  wherein a control cell line is substantially simultaneously tested.  
     
     
         3 . The method according to  claim 1  wherein the number of cells counted is about 40.  
     
     
         4 . The method according to  claim 1  wherein the number of cells counted is about 20.  
     
     
         5 . A method for determining the likelihood of cancer recurrence in a patient comprising the steps of: 
 (I) obtaining tumor cells from the patient;    (ii) applying to said cells a detectable probe capable of hybridizing to the HER-2/neu gene;    (iii) counting the number of HER-2/neu genes in cells; and    (iv) determining the likelihood of cancer recurrence wherein an average of at least about 4 HER-2/neu genes per tumor cell indicates a high likelihood of cancer recurrence.    
     
     
         6 . The method according to  claim 5  wherein at least one control cell line is substantially simultaneously tested.  
     
     
         7 . The method according to  claim 5  wherein the number of cells with HER-2/neu genes counted therein is no more than about 20.  
     
     
         8 . A kit of quality control cell lines for a HER-2/neu gene amplification detection system comprising: 
 (i) a non-amplified control cell line that exhibits a mean of about 3 or fewer HER-2/neu genes per cell; and    (ii) an amplified control cell line that exhibits a mean of about 10 or more HER-2/neu genes per cell.    
     
     
         9 . The kit according to  claim 8  and further comprising a low amplified control cell line that exhibits a mean of between about 3 and 10 HER-2/neu genes per cell.  
     
     
         10 . A kit of quality control cell lines for use with a breast cancer gene amplification detection system according to  claim 9  comprising: 
 (I) ATCC HTB 30 (SK-BR-3) cell line;  
 (ii) ATCC HTB 132 (MDA-MB-468) cell line; and  
 (iii) ATCC HTB 133 (T-47D) cell line.  
 
     
     
         11 . A kit for determining the number of copies of HER-2/neu genes in a cell comprising; 
 a) a detectable probe capable of hybridizing to the HER-2/neu gene; and    b) at least one control cell line selected from the group consisting of one that exhibits a mean of about 10 or more HER-2/neu genes per cell, one that exhibits a mean of between about 3 and 10 HER-2/neu genes per cell, and one that exhibits a mean of about 3 or fewer HER-2/neu genes per cell.    
     
     
         12 . The kit according to  claim 11  further comprising at least two of the control cell lines.  
     
     
         13 . The kit according to  claim 12  further comprising all three of the control cell lines.  
     
     
         14 . A method for preparing a standardized control slide comprising 
 mixing cells of a cell line in a liquid to form a suspension,    solidifying the suspension, whereby the cells are dispersed throughout the suspension to produce a solid form,    cutting thin sections from the solid form, and    mounting the thin section onto a slide.    
     
     
         15 . The method according to  claim 14  further comprising embedding the solid form in a solid matrix.  
     
     
         16 . The method according to  claim 15  wherein the solid matrix is paraffin.  
     
     
         17 . The method according to  claim 14  further comprising staining the section.  
     
     
         18 . The method according to  claim 14  further comprising preparing the section for binding an added ligand to a receptor in or on the cells.  
     
     
         29 . The method according to  claim 14  further comprising preparing the section so that nucleic acid in the cells is capable of hybridizing to an added solution containing a complementary nucleic acid probe.  
     
     
         20 . A quality control material for use as a standardized control comprising: 
 (a) a first cell line having an amplified HER-2/neu gene copy number; and    (b) a second cell line having a non-amplified HER-2/neu copy number;    wherein all of the cells in the quality control material are distributed throughout a solid medium in three dimensions.    
     
     
         21 . A thin section of a quality control material consisting essentially of a thin section cut from the material of  claim 20.