Differential immunohistochemical staining of cancerous cells with anti-single-stranded DNA antiserum in ordinary pathological paraffin section after DNA-denaturation by acid hydrolysis.
Based on the finding that the nuclear DNA of cancerous cells is much more unstable than that of non-cancerous cells, yielding a larger amount of single-stranded DNA by acid hydrolysis, we developed a new method of cancer cell detection in ordinary pathological sections by immunohistochemical staining with anti-single-stranded DNA antiserum after acid hydrolysis. Methylated bovine serum albumin was conjugated with heat-denatured calf thymus DNA and used as the antigen of single-stranded DNA, and white male rabbits were immunized with the antigen to obtain the polyclonal antiserum. Ordinary paraffin-embedded sections were prepared from the formalin-fixed biopsy specimens taken from 482 malignant tumors and 73 benign tumors of human epithelial and non-epithelial origins. Additional 82 biopsy specimens of borderline malignancy were also examined. The sections were immunohistochemically stained with the antiserum after RNase digestion and DNA denaturation by hydrolysis with 2 N HCl at 30 degrees C. The acid hydrolysis for 20-30 min was optimum for the specimens fixed with 10% buffered formalin at room temperature for 16-24 hrs, and all cancerous cells were specifically stained positive, in sharp contrast to the negative stainability of all non-cancerous cells including inflammatory cells. This new method gives us decisive help in making diagnosis of malignancy in daily pathological examination. The possibility of malignancy of borderline lesion was discussed.
Fukuda
M
M
Department of Pathology, Fukui Medical School, Japan.
Noriki
S
S
Imamura
Y
Y
Miyoshi
N
N
Kimura
S
S
Koizumi
K
K
Suzuki
K
K
Sakurai
H
H
Sakaguchi
R
R
D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology*; D002417:Cattle; D004273:DNA, Neoplasm / Q000276:immunology; D004277:DNA, Single-Stranded / Q000276:immunology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007106:Immune Sera; D007150:Immunohistochemistry / Q000379:methods*; D008297:Male; D009369:Neoplasms / Q000276:immunology / Q000473:pathology*; D009691:Nucleic Acid Denaturation; D016612:Paraffin Embedding; D011817:Rabbits
D000974:Antibodies, Antinuclear; D004273:DNA, Neoplasm; D004277:DNA, Single-Stranded; D007106:Immune Sera