Comparison of in situ methods to assess DNA cleavage in apoptotic cells in patients with breast cancer.
BACKGROUND
Apoptosis has a role in many cellular processes including development, normal tissue homeostasis, and malignancy. This aspect of research is relatively new with distinct methods of analysing disparate biochemical and genetic events to measure apoptotic cells. The use of biotinylated nucleotides to identify DNA strand breaks is a commonly reported method of estimating cells numbers undergoing apoptosis; however, investigators report inconsistent results for a variety of reasons.


OBJECTIVE
To compare two in situ techniques of measuring apoptosis: in situ nick translation (ISNT) and TdT mediated dUTP-biotin nick end labelling (TUNEL); and to assess DNA cleavage in 20 paired paraffin wax embedded breast cancer tissues from patients; one group who had received no prior treatment and one group who had received chemohormonal treatment.


CONCLUSIONS
Apoptotic scores obtained from paraffin wax embedded human breast cancer after using ISNT and TUNEL methods were not significantly different (p = 0.11). A strong correlation between scores obtained from the two techniques was found (r = 0.758, p < 0.0001). Optimisation of both techniques is crucial to ensure maximal assay performance in breast cancer tissue.
Mainwaring
P N
PN
Academic Department of Biochemistry, Royal Marsden Hospital, London, UK. paul@tcr.ac.uk
Ellis
P A
PA
Detre
S
S
Smith
I E
IE
Dowsett
M
M
D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D017209:Apoptosis; D019921:Biotinylation; D001943:Breast Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology*; D018270:Carcinoma, Ductal, Breast / Q000188:drug therapy / Q000235:genetics / Q000473:pathology*; D053938:DNA Fragmentation; D004273:DNA, Neoplasm; D003857:Deoxyuridine; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D016612:Paraffin Embedding
D004273:DNA, Neoplasm; D003857:Deoxyuridine