PMID- 2424870
OWN - NLM
STAT- MEDLINE
DCOM- 19860725
LR  - 20190828
IS  - 0301-5564 (Print)
IS  - 0301-5564 (Linking)
VI  - 84
IP  - 4-6
DP  - 1986
TI  - Different instability of nuclear DNA at acid hydrolysis in cancerous and 
      noncancerous cells as revealed by fluorescent staining with acridine orange.
PG  - 556-60
AB  - Ehrlich cancer cells and inflammatory cells in mouse ascitic fluid were 
      hydrolyzed and stained with acridine orange (AO). The AO hydrolysis curves for 
      G1/G2 + M phase cancer cells and inflammatory cells were differentially 
      determined using flow cytometry by monitoring the metachromatic red-shifted 
      fluorescence of the fluorochrome bound to the single-stranded DNA produced by 
      acid hydrolysis. By computer fitting of the Bateman function to the hydrolysis 
      curves, the kinetic parameters k1 (rate constant for the production of 
      single-stranded DNA), k2 (rate constant for the degradation of the produced 
      single-stranded DNA), and y0 (theoretical value of the single-stranded DNA 
      present initially) were determined. It was found that the k2 value, which 
      reflects the degree of DNA instability, was much higher for cancer cells in both 
      the G1 and G2 + M phases than for inflammatory cells. This finding led us to 
      develop a method for the differential AO staining of cancer cells and 
      non-cancerous cells utilizing the different degree of DNA instability at acid 
      hydrolysis. AO staining after hydrolysis with 2N HCl at 30 degrees C for 8.5 min 
      was found to be the optimal method. In the 60 cases of human malignant epithelial 
      and nonepithelial tumors tested, all of the malignant tumor cells emitted 
      metachromatic red fluorescence, while all of the nonmalignant tumor cells (5 
      cases of benign tumor) and normal cells emitted orthochromatic green fluorescence 
      when observed with a violet excitation light under a fluorescence microscope. 
      This new technique can be a useful tool for the screening of malignancy in 
      exfoliative cytology and also for basic cancer research.
FAU - Fukuda, M
AU  - Fukuda M
FAU - Miyoshi, N
AU  - Miyoshi N
FAU - Hattori, T
AU  - Hattori T
FAU - Sugihara, H
AU  - Sugihara H
FAU - Hosokawa, Y
AU  - Hosokawa Y
FAU - Nakanishi, K
AU  - Nakanishi K
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Histochemistry
JT  - Histochemistry
JID - 0411300
RN  - 0 (DNA, Neoplasm)
RN  - 9007-49-2 (DNA)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - *Acridine Orange
MH  - Animals
MH  - Cell Cycle
MH  - DNA/*analysis
MH  - DNA, Neoplasm/*analysis
MH  - Humans
MH  - Hydrolysis
MH  - Male
MH  - Mice
MH  - Microscopy, Fluorescence
MH  - Neoplasms/*diagnosis
MH  - Staining and Labeling
EDAT- 1986/01/01 00:00
MHDA- 1986/01/01 00:01
CRDT- 1986/01/01 00:00
PHST- 1986/01/01 00:00 [pubmed]
PHST- 1986/01/01 00:01 [medline]
PHST- 1986/01/01 00:00 [entrez]
AID - 10.1007/BF00482991 [doi]
PST - ppublish
SO  - Histochemistry. 1986;84(4-6):556-60. doi: 10.1007/BF00482991.