Nuclear morphometry and DNA flow cytometry as prognostic methods for endometrial carcinoma

Abstract

In a series of 227 women with endometrial carcinoma stages I-IV the prognostic value of nuclear morphometry and DNA analysis was evaluated prospectively. The tumors were also classified according to histologic subtype, degree of differentiation (FIGO), and nuclear grade. The DNA analysis (ploidy and S-phase fraction) was made using flow cytometry on fresh-frozen tissue. The morphometric measurements and the grading of the tumors were done on both fresh-frozen tissue and on formalin-fixed and paraffin-embedded tissue. The rate of recurrence for the complete series was 14% with 2.2% vaginal metastases. The overall 5-year survival rate was 68% and the corrected survival rate 76%. The minimum nuclear diameter and the standard deviation (SD) of the maximum nuclear diameter were significant and independent prognostic variables in a Cox multivariate analysis when analyzed on paraffin-embedded tissue. The nuclear grade was the single most prognostically informative variable. A corresponding analysis on fresh-frozen tissue showed that the S-phase fraction and the FIGO grading of the tumor were significant variables. The morphometric variables and the nuclear grading were non-significant prognostic variables. Morphologic changes in tumor cell nuclei during the freezing and thawing procedures may explain the loss of prognostic information. A combination of DNA index (fresh-frozen tissue) and the mean values of the shortest nuclear diameter and the SD of the longest diameter (paraffin-embedded tissue) gave the best prognostic information vis-a-vis cancer-related death in an all-possible-subsets regression analysis.

Prognostic impact of morphometric nuclear grade of endometrial carcinoma

“…Van der Putten et al 27 reported an independent prognostic impact of nuclear morphometry in a small series of Stage I endometrial carcinoma patients. In a larger study examining only curettage specimens due to preoperative radiation therapy, a prognostic impact of nuclear morphometry was reported by Sorbe et al 28 Our current results are in accordance with previous findings, although nuclear perimeter was found to be prognostically the most powerful in our series compared with DMIN in the earlier reports. 27,28 However, all variables expressing nuclear size had a prognostic impact in our study.…”

Confirmation of the prognostic value of the ECPI-1 score (myometrial invasion, DNA-ploidy and mean shortest nuclear axis) in FIGO stage I endometrial cancer patients with long follow-up

“…6). Sorbe et al (22) also found that myometrial invasion DNA-ploidy, %S-phase cells and the mean shortest nuclear axis, were prognostically the most significant, thus confirming the value of the essential features of the ECPI-1. Abeler et al (23) found that the prognostic value of vascular invasion was limited.…”

Comparison of clinical staging of benign and malignant ovarian tissues with DNA flow cytometry

“…This finding is well supported in the literature [39,40,50,52,64] . The presence of this atypical DNA suggests a prognosis that is poorer than a sample with normal DNA [6,8,11,13,14,16,24,39,41,43,52,59,60,66,70] . The aneuploid characterization of these benign tumors should at least require a closer, independent pathological examination.…”

“…Papers that do not publish their coefficients of variation and do not show representative DNA histograms should be reviewed with caution [39,40,[43][44][45][46] . Furthermore, preparation techniques using formalin-fixed, paraffinated tissues that use a low pH of 2.0 and non-DNA specific staining methods, such as propidium iodide, are less than ideal in performing the proper sample preparation [47] .…”

“…We have been pioneers in developing better ways to measure DNA in tissues [39,40,[43][44][45][46] . The purpose of the research presented here is to show the best procedures for measuring DNA in nuclei derived from paraffinated tissues.…”