Nuclear DNA cytophotometry in prostate carcinoma

Abstract

Cytologic evaluation of aspired samples of palpable nodules in the human prostate developed into a routine method, since Esposti (5) classified carcinoma with high (Gl), moderate (G2), and poor (G3) differentiation and evaluated the prognostic importance.The malignant cells frequently have a different DNA content than do normal cells (1,3,7,8,13). The purpose of this study was to determine the DNA content of prostate cancer cells before and during endocrine therapy, to compare it with the cytologic grading, the clinical state, and to evaluate possible prognostic factors. MATERIALS AND METHODSEighty patients with freshly diagnosed prostatic cancer underwent transrectal fine-needle aspiration biopsy prior to treatment (6). The biopsies were taken fanlike from the suspected area and neighboring unsuspected areas. Later repeat biopsies were taken from the same tumor region at 6-(n = 76) and 12-month intervals (n = 70) in surviving patients. The corresponding clinical state of complete or partial remission, stable disease, or progression was defined according to Schmidt et a1 (9).It was judged by digital rectal examination, percutaneous transvesical sonography, serum acid phosphatase activity, and radionuclide bone scan, followed by skeletal radiography in positive cases. Twenty patients with benign prostatic hypertrophy served as controls.The biopsied specimen was used for cytologic morphologic grading (5) and for single-cell fluorescence photometry (10-121, performed with the microscope photometer MPV 2 (Leitz, Wetzlar, West Germany).The samples were fixed with Merckofix spray and stored at +4-+8"C for the staining procedure. The samples were fixed again for 30 min in 96% ethyl alcohol and hydrolyzed in a thermostatic-controlled water bath at +28 f 0.2"C in 4N hydrochloric acid. Then 0.01% acriflavine-Schiff (Serva, Heidelberg, West Germany) was used for nuclear staining at room temperature for 1 hour. The samples were washed in SO2 solution, dehydrated in alcoholic solution, immersed in xylol for 5 min, and covered with nonfluorescent Cargille Oil (Cargille Laboratories, Cedar Grove, NY). This oil had a nucleuslike refractive index of nk0 = 1.54.About 100 cells were measured in every preparation. Normal prostate cells and leukocytes were used for evaluation of the normal diploid 2c DNA content. The values were displayed as histograms and analyzed statistically with the median-quartile test (2), because the DNA values differed in most cases from a normal distribution.Five parameters were extracted from the distribution of the DNA values and were used for the analysis. These parameters were the minimum, first quartile, median, third quartile, and maximum. A sixth parameter was Z, the 2c value, which was defined as the percentage of cells with a DNA content of 2c f 25% (12). RESULTSThe pretherapeutic DNA histograms had single peaks, were bimodal, or showed a broadly scattered DNA disAddress reprint requests to Dr.

Image analysis and grading of prostate adenocarcinoma

“…In 1984 Seppelt et al [74] reported good correlation between clinical follow up and progressive changes in the DNA histogram, as obtained by static cytophotometry on FNA material. However, progressive changes of the histogram were seen in more than half the cases with clinical remission.…”

DNA analysis by flow cytometry of paraffin embedded core biopsies of the prostate

“…We were interested in determining if the information generated by DNA analysis, reflecting mostly prostatectomy tissue, could also be obtained by flow cytometry before prostatectomy from the very small, multiple, core biopsies of the prostate divided into normal segments versus segments containing malignant glands. Although there are some reports using the DNA measurement methods of image cytometry from fine needle aspirate (FNA) material [9,10], as well as flow cytometry to measure the DNA content of fresh FNA material [11,12] or freshly minced core biopsies [13,14] of the prostate, we are not aware of any reports utilizing flow cytometry to analyze fixed, paraffin-embedded biopsies. One potential benefit of using the fixed, paraffin-embedded core biopsies, instead of fresh tissue when using flow cytometry, is the technical advantage of being able to select and analyze different areas of the core biopsies enriched for malignant versus benign cells.…”

New Prognostic Factors in Prostatic Carcinoma

“…A more favorable response of tumors to endocrine therapy and a longer progression-free and over all survival rate of patients receiving such therapy have been observed in diploid and tetraploid cases than in triploid or hypertetraploid cases [64,65,71]. Histological grade or clinical stage did not predict the response to endocrine therapy [65,71], New aneuploid peaks often appear in nonresponsive tumors [77] suggesting that the evolvement of tumor karyotype may have contributed to the disease progression. Ekman et al [66] showed that DNA aneuploid prostatic tumors were more often andro gen-receptor-negative than diploid tumors.…”