Nuclear DNA of the gallbladder cancer was stained by the Feulgen method and analyzed using a new concept of DNA score. DNA score was determined by the peak ploidy unit and decided numbers of frequency in each ploidy unit. As the result, a significant correlation between DNA score and the depth of cancer invasion was demonstrated. The depth of cancer invasion, as reported already, has close correlation to the prognosis of the patients of gallbladder cancer. These results indicate that the nuclear DNA analysis is useful for the determination of malignancy and the prediction of the prognosis of the patients. T gallbladder cancer ; nuclear DNA ; depth of cancer invasion ; Feulgen stainingThe authors have already reported that, in carcinoma of the gallbladder, the prognosis of the patients with slight depth of cancer invasion was comparatively good (Sato et al. 1981). A correlation between the depth of cancer invasion and nuclear DNA pattern of cancer cells, one of the indicators of malignancy, was investigated in cancer of the gallbladder.Materials and methods. Twenty-six patients with gallbladder cancer resected curatively in our department from 1960 to 1983 were investigated. The depth of cancer invasion of these patients were mucosa (m) in 5 cases, proper muscle (pm) in 4, subserosa (ss) in 4 and serosa (s) in 13.Surgical specimens of the gallbladder cancer, fixed in 10% formalin and embedded in paraffin in usual procedures, were sectioned at 6 pm and stained by the Feulgen method to demonstrate DNA (Leuchtenberger 1958). Nuclear DNA content of 100 cancer cells at the periphery of the cancer nests, verified in hematoxylin and eosin-stained sections, was measured using an OLYMPUS microspectrophotometer.Frequency histograms were plotted from the values obtained and were expressed in ploidy unit, in which the mean value of DNA content of 20 lymphocytes in the same section was taken as the diploid value (2C. C = DNA content of a haploid chromosome set).Results and comments. In histogram, the peak of frequency was observed in the ploidy unit of 2.0 to 2.50 in 2 cases, 2.5 to 3.00 in 21 cases and 3 to 3.50 in 3 cases. Fig. 1 shows the histogram of average frequency of nuclear DNA of the patients with the respective depth of cancer invasion. DNA patterns can be classified according to the ploidy unit showing the peak of frequency in histogram as well as to the percent value of frequency in each ploidy unit. Since nuclei having a large ploidy unit contain a large amount of chromosome, which suggests high malignancy, the frequency value in large ploidy