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

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.     

Nuclear DNA content, proliferative activity, and p53 expression related to clinical and histopathologic features in endometrial carcinoma

Abstract. Lundgren C, Auer G, Frankendal B, Moberger B, Nilsson B, Nordström B. Nuclear DNA content, proliferative activity, and p53 expression related to clinical and histopathologic features in endometrial carcinoma.
The purpose of this study was to evaluate the prognostic impact of image cytometry DNA ploidy, MIB-1, and p53 in relation to clinicopathologic variables in 376 consecutive patients with endometrial carcinoma stages I–IV. Following primary treatment 358 patients were considered tumor-free. Relapses and tumor-specific deaths of these patients were noted. Image cytometry DNA ploidy ( n = 340) and expression of MIB-1 ( n = 318) and p53 ( n = 323) were studied. In univariate analysis, stage ( P < 0.001), histopathologic subtype ( P < 0.001), degree of differentiation ( P < 0.001), HRT ( P = 0.034), DNA ploidy ( P < 0.001), and p53 ( P < 0.001) were significant predictors of relapse. Patient age showed that the estimated mean risk of relapse increases with nearly 64% per decade in life ( P 0.003), and the MIB-1 expression with 21% per 10-unit increment ( P 0.004). In multivariate analysis, degree of differentiation, MIB-1, and p53 lost their prognostic capability. However, after stage and histopathologic subtype, image cytometry DNA ploidy was the strongest predictor of outcome and was of value in predicting the risk for relapse. The combination of DNA ploidy, MIB-1, and p53 expression was an even stronger predictor of relapse-free survival than the individual prognostic factors.     

Significance of morphometric, DNA cytometric features, and other prognostic markers on survival of endometrial cancer patients in northern Norway

Abstract. Ørbo A, Rydningen M, Straume B, Lysne S. Significance of morphometric, DNA cytometric features, and other prognostic markers on survival of endometrial cancer patients in northern Norway.
The objective of this study was to evaluate the prognostic value of nuclear morphometric features and DNA ploidy by flow cytometry next to depth of myometrial invasion and vascular invasion in endometrial cancer of all FIGO stages.
A total of 123 women (103 FIGO stage I, eight stage II, and 12 stage III and IV) from northern Norway were studied. The follow-up period was between 7 and 19 years. The median age of patients was 62 years. The primary surgery was performed in the University Hospital of Tromsø or in the seven different reference hospitals in the northern part of Norway after an endometrial cancer diagnosis. The histologic, morphometric, flowcytometric and immunohistochemical investigations were based on archival paraffin-embedded material. The information regarding the follow-up data and clinical information were obtained from the medical records.
Thirteen (10.6%) patients from the entire group (all stages) but only three (2.7%) of the FIGO stage I and II patients died from locally recurrent or metastatic disease. FIGO substage ( P = 0.0006; odds ratio [OR] = 16.44, 95% confidence interval [CI] = 3.36–80.45), vascular invasion ( P = 0.01, OR = 6.42, CI = 1.57–26.34) and nuclear size ( P = 0.025, 0 R = 1.3, CI = 1.05–1.61) were independently correlated with recurrence in a multivariate analysis but histologic grade and DNA ploidy were not. Vascular invasion was poorly reproducible both between and within the same observer, however.
In this retrospective study of all stages of endometrial carcinoma with long follow-up periods the primary tumor characteristics nuclear perimeter and FIGO stage were of prognostic significance in addition to the poorly reproducible vessel invasion.     

Nuclear shape: An independent predictor of survival in patients with ovarian carcinoma

Geisler JP, Geisler HE, Wiemann MC, Givens SS, Zhou Z, Miller GA. Nuclear shape: An independent predictor of survival in patients with ovarian carcinoma. Int J Gynecol Cancer 1998; 8 : 164–167.
Since nuclear morphometry has recently been shown to be of prognostic value in several malignancies, including endometrial cancer, the authors attempted to see if those same morphometric features of nuclear size, shape and summed optical density had an impact on survival in patients with epithelial malignancies of the ovary. Eighty-three consecutive patients with epithelial malignancies of the ovary had their tumors studied in a quantitative manner evaluating nuclear size (NUSZ), shape (NUSH), and summed optical density (NUSD). Touch preps from this tissue were made. Patient records were examined for FIGO stage, grade, histology, as well as level of cytoreduction. The mean follow-up was 37 months (median 30 months, range 24–55 months). Multivariate analysis proved that the level of cytoreduction ( P = 0.0002), FIGO stage ( P = 0.025), and NUSH ( P = 0.036) were independent predictors of survival. NUSH ( P = 0.018) and NUSD ( P = 0.020) were significantly different among the different histologic grades. Additionally, NUSH ( P = 0.007) and NUSD ( P = 0.001) were significantly different between patients who did and did not survive. NUSZ was not significantly related to survival, stage, grade or level of cytoreduction. In conclusion, NUSH and NUSD both appear to be important morphometric features in epithelial ovarian carcinoma. Further study concerning their applicability as prognostic factors is warranted.     

On mononuclear cells in human endometrial carcinoma

The aim of this study was to evaluate the local immune reactivity in patients with untreated endometrial carcinoma. The immune assay included immunostaining of mononuclear cells in cryo- and/or paraffin sections in 19 untreated endometrial cancers and in 23 normal endometrial tissues as controls. In several cases Ficolli preparations after mechanical dispersion were successfully made with cytospin and/or cytofluorometric study of the mononuclear cells. Throughout the normal menstrual cycle as well as the postmenopausal, both T-lymphocytes and macrophages appeared in the stroma, in aggregates or diffusely scattered. B-cells were rare. During the secretory phase the highest cell number occurred, especially T-helpers (CD4) were increased. Endometrial carcinomas had higher mean values of lymphocytes (CD3, CD4 and CD8) and IL-2 receptor positive cells than normal endometrial controls. This increase of lymphocytes – and also macrophages – was related to the degree of malignancy. C57 positive natural killer (NK) cells, how-ever, were practically absent in the malignant tissue. This immunogenic activity was much stronger in endometrial cancer than that found in ovarian cancer.     

Denser trabecular bone in women with endometrial carcinoma

Bone mineral density (BMD) was measured in the distal radius of patients with endometrial carcinoma (EC). The patients were classified into two subgroups depending on whether earlier hormonal replacement therapy (HRT) was given. Two groups of women were recruited as controls: patients with post-menopausal bleeding for non-malignant reasons (hospital controls) and healthy women, free of gyn-ecologic symptoms (non-hospital controls). The BMD was significantly higher in the cancer patients and also in the hospital controls than in the non-hospital controls. When several possible confounding factors were checked for in a multivariate analysis, BMD still differed between the groups. This could lend support to the hypothesis that patients with EC may have an altered endogenous endocrine status which eventually affects their bone mass. The results also stress the importance of using strictly defined, healthy women as controls.     

Surgical cytoreduction in stage IV endometrioid endometrial carcinoma

Objective

To evaluate the role of surgical cytoreduction and the amount of residual disease in patients with newly diagnosed stage IV endometrioid endometrial carcinoma (EC).

Methods

Patients with stage IV EC of endometrioid histology who underwent surgery at our institution from 1977 to 2003 were identified. Patients with microscopic stage IV disease were excluded. Progression-free survival (PFS) and overall survival (OS) were estimated using Kaplan Meier method and compared with log-rank test.

Results

A total of 58 patients were identified, of which 9 (15.5%) had no gross residual (NGR) after surgery, 11 (19.0%) had residual disease ≤ 1 cm, 32 (55.1%) had residual disease > 1 cm, and 6 (10.3%) had no cytoreduction attempted. The median PFS was 11.1 months (95% CI, 9.8-12.3) and the median OS was 19.2 months (95% CI, 8.5-29.9) for the cohort. The median PFS was 40.3 months (95% CI, 0-93.9) for patients with NGR disease, 11 months (95% CI, 9.9-12.1) for patients with any residual disease, and 2.2 months (95% CI, 0.1-4.2) for patients who did not have attempted cytoreduction (P < 0.001). The median OS was 42.2 months (95% CI, not estimable) for patients with NGR disease, 19 months (95% CI, 13.9-24.1) for patients with any residual disease, and 2.2 months (95% CI, 0.1-4.2) for patients that did not have attempted cytoreduction (P < 0.001).

Conclusion

Though stage IV endometrioid EC has a poor prognosis, surgical cytoreduction to no gross residual disease in a highly select group of patients is associated with improved survival.     

Predicting pelvic lymph node metastasis in endometrial carcinoma

BACKGROUND: To determine the possibility of individualizing the pelvic lymph node dissection in patients with endometrial cancer, the relationship between pelvic lymph node (PLN) metastasis and various prognostic factors was retrospectively investigated. METHODS: From 1979 to 1994, 175 patients with endometrial carcinoma were treated with either total or radical hysterectomy combined with a PLN dissection as initial therapy. The prognostic factors examined included clinical stage, patient age, histological grade, the microscopic degree of myometrial invasion (DMI), cervical invasion, adnexal metastasis, and macroscopic tumor diameter (TD). RESULTS: Of the 175 patients undergoing PLN dissection, 24 (14%) had PLN metastasis. An endometrial cancer with PLN metastasis had a significantly longer diameter than those without PLN metastasis. The frequency of PLN metastasis increased along with increases in tumor diameter. A logistic regression analysis revealed DMI and TD to be independently correlated with PLN metastasis. The formula based on the coefficients of TD and DMI obtained from the analysis also showed a good correlation, which allowed us to estimate the probability of patients having PLN metastasis. CONCLUSIONS: DMI and TD could accurately estimate the status of PLN in endometrial carcinoma patients.     

Pretreatment hemoglobin, platelet count, and prognosis in endometrial carcinoma

Abstract. Tamussino K, Gücer F, Reich O, Moser F, Petru E, Scholz HS. Pretreatment hemoglobin, platelet count, and prognosis in endometrial carcinoma.
We analyzed whether a low preteatment hemoglobin level is a prognostic factor in endometrial cancer and whether it is associated with thrombocytosis. Two hundred and twelve patients with endometrial cancer treated with surgery were reviewed. Data were analyzed with Pearson's chi-squared test, Fisher's exact test in contingency tables, the Mann–Whitney U -test, the Student's t -test, and Kaplan-Meier estimates. Multivariate analysis was performed with the log-rank test and the Cox proportional hazard model. Thirty-nine patients (18%) had a pretreatment hemoglobin value of <12.0 g/dL. These 39 patients had significantly higher rates of nonendometrioid histology, high-grade tumors, myometrial invasion of >50%, adnexal involvement, lymph-vascular space involvement, and advanced FIGO stage than patients with hemoglobin ≥12.0 g/dL. The rate of thrombocytosis was significantly higher in patients with a low hemoglobin level (36% vs. 8% , P < 0.01). The overall 5-year survival rate of patients with low pretreatment hemoglobin was 59% compared with 89% for those with hemoglobin ≥12 g/dL ( P < 0.01). In the multivariate analysis age, thrombocytosis, nonendometrioid histology, high-grade histology, and advanced FIGO stage were significantly associated with a poor prognosis whereas adnexal involvement, lymph-vascular space involvement, low hemoglobin and myometrial invasion were not. These data indicate that low pretreatment hemoglobin is a prognostic factor in patients with endometrial cancer and that it is associated with thrombocytosis. Low hemoglobin was strongly associated with other unfavorable prognostic factors so that it was significant in the univariate but not the multivariate analysis.     

Resection of lymph node metastases influences survival in stage IIIC endometrial cancer

OBJECTIVE: Surgical staging of endometrial cancer identifies those patients with microscopic metastatic disease most likely to benefit from adjuvant therapy and may also confer therapeutic benefit. Our objective was to compare survival of patients who underwent resection of grossly positive lymph nodes (LN) to those with microscopically positive LN. METHODS: Patients had stage IIIC endometrial cancer with pelvic and/or aortic LN metastases and underwent surgery between 1973 and 2002. Exclusion criteria included pre-surgical radiation and second primary cancer. Survival was analyzed using Kaplan-Meier method and Cox proportional hazards model. RESULTS: Mean age of 96 patients with stage IIIC endometrial cancer was 64. There were 45 cases with microscopic LN involvement and 51 with grossly enlarged LN. Overall, 41% had disease in aortic LN, which in 18% represented isolated aortic LN metastasis. Adjuvant therapies were given to 92% of patients (85% radiotherapy, 10% chemotherapy, 10% progestins). Among those with grossly involved LN, 86% were completely resected. Five-year disease-specific survival (DSS) was 63% in 45 patients with microscopic metastatic disease compared to 50% in 44 patients with grossly positive LN completely resected and 43% in 7 with residual macroscopic disease. In multivariable analyses, gross nodal disease not debulked (HR=6.85, P=0.009), serosal/adnexal involvement (HR=2.24, P=0.036), diagnosis prior to 1989 (HR=4.33, P<0.001), older age (HR=1.09, P<0.001), and >2 positive lymph nodes (HR=3.12, P=0.007) were associated with lower DSS. CONCLUSION: Grossly involved LN can often be completely resected in patients with stage IIIC endometrial cancer. These retrospective data provide evidence suggestive of a therapeutic benefit for lymphadenectomy in endometrial cancer.     

Metformin use and endometrial cancer survival

Objective

Impaired glucose tolerance and diabetes are risk factors for the development of uterine cancer. Although greater progression free survival among diabetic patients with ovarian and breast cancers using metformin has been reported, no studies have assessed the association of metformin use with survival in women with endometrial cancer (EC).

Methods

We conducted a single-institution retrospective cohort study of all patients treated for uterine cancer from January 1999 through December 2009. Demographic, medical, social, and survival data were abstracted from medical records and the national death registry. Overall survival (OS) was estimated using Kaplan–Meier methods. Cox models were utilized for multivariate analysis. All statistical tests were two-sided.

Results

Of 985 patients, 114 (12%) had diabetes and were treated with metformin, 136 (14%) were diabetic but did not use metformin, and 735 (74%) had not been diagnosed with diabetes. Greater OS was observed in diabetics with non-endometrioid EC who used metformin than in diabetic cases not using metformin and non-endometrioid EC cases without diabetes (log rank test (p = 0.02)). This association remained significant (hazard ratio = 0.54, 95% CI: 0.30–0.97, p < 0.04) after adjusting for age, clinical stage, grade, chemotherapy treatment, radiation treatment and the presence of hyperlipidemia in multivariate analysis. No association between metformin use and OS in diabetics with endometrioid histology was observed.

Conclusion

Diabetic EC patients with non-endometrioid tumors who used metformin had lower risk of death than women with EC who did not use metformin. These data suggest that metformin might be useful as adjuvant therapy for non-endometrioid EC.     

Obesity, diabetes, and other factors in relation to survival after endometrial cancer diagnosis

Endogenous and exogenous sources of estrogen and characteristics altering these hormone levels have been related to endometrial cancer risk; however, their relationship to survival following diagnosis is less clear. In a population-based study, we examined whether mortality after endometrial cancer diagnosis was affected by prediagnosis obesity, diabetes, smoking, oral contraceptive use, parity, or postmenopausal hormone (PMH) use. Eligible women, aged 40-79 years, diagnosed from 1991-1994 with incident invasive endometrial cancer and identified through the Wisconsin statewide mandatory cancer registry were invited to participate. Of 745 eligible cases, 166 women were deceased after 9.3 years of follow-up, with 43 attributable to endometrial cancer, based upon vital records linkage. Hazard rate ratios (HRR) and 95% confidence intervals were adjusted for age at diagnosis, menopausal status, stage of disease, and other exposures of interest. Obese women (body mass index [BMI] >or=30 kg/m(2)) prior to endometrial cancer diagnosis had an increased risk of both all-cause (HRR=1.6, 95% CI 1.0-2.5) and endometrial cancer (HRR=2.0, 95% CI 0.8-5.1) mortality, compared with nonoverweight women (BMI<25 kg/m(2)). Endometrial cancer cases with diabetes also had an increased risk of all-cause mortality compared with nondiabetic women (HRR=1.7, 95% CI 1.1-2.5), although there was no association with endometrial cancer mortality. There were no associations between PMH use, oral contraceptive use, parity, or smoking and mortality from any cause. The results suggest that history of obesity and diabetes may increase risk of mortality after endometrial cancer diagnosis; modification of these characteristics may improve survival after endometrial cancer diagnosis.     

Phase II study of fulvestrant in recurrent/metastatic endometrial carcinoma: a Gynecologic Oncology Group study

Objectives.

To evaluate the activity and toxicity of fulvestrant in advanced, recurrent, or persistent endometrial carcinoma.

Methods.

Eligible patients with advanced, recurrent or persistent endometrial carcinoma not amenable to curative therapy were treated with fulvestrant at a dose of 250 mg by IM injection every 4 weeks for at least 8 weeks. Therapy was continued until evidence of progressive disease, or adverse effects prohibited further therapy. Response was assessed in patients with at least one target lesion as defined by Response Evaluation Criteria in Solid Tumors (RECIST) v1.0. Immunohistochemical analysis of tumor tissue (histology or cytology) for estrogen and progesterone receptors was required from the metastatic or recurrent site.

Results.

Sixty-seven patients were enrolled in this study. Upon review, 14 patients were excluded. In the 22 estrogen receptor (ER) negative patients, no patients demonstrated either a complete or partial response, and 4 (18%) demonstrated stable disease (as best response). In the 31 ER positive patients, 1 (3%), 4 (13%) and 9 (29%) patients demonstrated a complete, partial response, and stable disease (as best response), respectively. The median progression free survival and overall survival in the ER negative patients were 2 and 3 months and in the ER positive patients 10 and 26 months. Treatment was well tolerated, and no patient discontinued therapy due to toxicity.

Conclusions.

Fulvestrant has minimal activity in advanced, recurrent, or persistent endometrial carcinoma.     

Impact of hysteroscopy on disease-free survival in clinically stage I endometrial cancer patients

Abstract. Obermair A, Geramou M, Gücer F, Denison U, Graf AH, Kapshammer E, Medl M, Rosen A, Wierrani F, Neunteufel W, Frech I, Preyer O, Speiser P, Kainz C. Impact of hysteroscopy on disease-free survival in clinically stage I endometrial cancer patients.
Recent data strongly suggest tumor cell dissemination of endometrial carcinoma cells in the course of fluid hysteroscopy. In patients who had endometrial cancer which was (except for peritoneal cytology) confined to the uterus, the disease-free survival (DFS) of 135 patients who underwent hysteroscopy prior to staging laparotomy was compared with the DFS of 127 patients without hysteroscopy. After a median follow-up of 23 months, 10 patients experienced tumor recurrence. Although there was a trend towards a higher incidence of positive peritoneal cytology at laparotomy in patients who underwent hysteroscopy, this difference did not achieve statistical significance ( P = 0.47). For 5 years, the DFS was 92.4% in patients with hysteroscopy and 84.7% in patients without hysteroscopy before laparotomy (log-rank, P = 0.782). Our data therefore suggest a similar short-term DFS in endometrial cancer patients with and without hysteroscopy prior to laparotomy.     

DNA-content in endometrial carcinoma of tamoxifen-treated breast cancer patients

DNA measurements and histopathologic evaluation were performed in 17 patients treated with adjuvant tamoxifen for early breast cancer and who developed endometrial carcinoma during or after the tamoxifen therapy. The tumors were exclusively characterized by euploid DNA content except for two cases, one mixed mesodermal sarcoma, a highly malignant and rare tumor, and one adenocarcinoma. Although the use of adjuvant tamoxifen therapy most likely enhances the risk of developing endometrial carcinoma, the beneficial effects of adjuvant breast cancer treatment is of well-known clinical importance. The hazards of giving long-term tamoxifen seem to be low since the endometrial tumors were associated with low-grade malignancy and euploid DNA pattern.