Expression of matrix metalloproteinase enzymes in endometrium of women with abnormal uterine bleeding.

OBJECTIVES: Abnormal uterine bleeding (AUB) is caused by derangement of physiological processes of tissue growth, shedding and regeneration. It is known that interplay between metalloproteinases (MMP's) and tissue inhibitors of metalloproteinases (TIMP's) may play a crucial role in its occurrence. AIM: To define if expression of proMMP-2, MMP-2 and TIMP-1 in endometrium of women with AUB is dependent on steroid sex hormone concentration and histopathological picture. MATERIALS AND METHODS: Endometrial scraps were taken from 21 women with AUB and 19 controls. Samples were evaluated in light microscopy by a certified pathologist. Activity of proMMP-2 and MMP-2 proteins levels were evaluated by gelatin zymography and TIMP-1 by reversed zymography. The results has been correlated with serum estradiol and progesterone concentrations in linear regression model. RESULTS: Expression: of proMMP-2 in endometrium of women with AUB is correlated with estradiol concentration and inversely correlated with progesterone levels. It was significantly higher in women with dysfunctional endometrium (p<0.001). Expression of MMP-2 was highest in women with endometrial polyps and longer bleeding (p<0.01), while expression of TIMP-1 was independent from hormone concentration. CONCLUSION: Lack of correlation between proMMP-2 and MMP-2 levels suggest different pathway of their activation in AUB. ProMMP-2 is up regulated by estradiol and down regulated by progesterone while MMP-2 levels increase with the length of bleeding.

The activity of mitochondrial superoxide dismutase in normal and pathological endometrium and correlation with the hormonal status of women.

GOAL: The goal of the study is to demonstrate the activity of superoxide dismutase mitochondria (MnSOD-e) in normal and pathologic endometrium and correlation of hormonal status of these cases with the MnSOD-e activity. MATERIALS AND METHODS: We analyzed 70 female patients, of which 30 of them had bleeding from the uterus (Group A) and 40 women had bleeding from the uterus, as well as a confirmed histopathological diagnosis of endometrial hyperplasia or endometrium carcinoma (Group B). In the follow-up we analyzed: age (respondents divided into five categories), parity (without pregnancy and multiple pregnancies), ultrasound (to determine whether there are pathological changes in the small pelvis), hormonal status of women (we took the blood of subjects to determine follicle stimulating hormone-FSH, luteinizing hormone-LH, progesterone-Pr, estradiol-Es), histopathological analysis (the material was collected by exploratory curettage of normal and pathologically altered endometrium), determining the activity of antioxidant enzymes in the blood and endometrium (we determined the activity of MnSOD-e, whose activity was determined in normal and pathological endometrium). RESULTS: Within age groups dominated patients from 41-50 years, as well as multiple pregnancies. In the experimental group the mean results had lower values of the MnSOD enzyme in blood (0.93) and endometrium (1.94) as compared to the control group in blood (1.27), and endometrium (2.79). The MnSOD-e levels in the follicular phase was approximately at the same level in the experimental and control group, while the values in the luteal phase and stage of menopause was greater in control compared to the experimental group. MnSOD-e levels in endometrium in the follicular phase and stage of menopause were lower in experimental than in the control group, whereas in the luteal phase in the experimental group the value was higher than in the control group. CONCLUSIONS: On the basis of the results, which show a decrease in MnSOD activity both in the blood and endometrium of patients with hyperplasia and endometrium carcinoma we can see the importance of detecting activity of these enzymes in the diagnosis of the mentioned histological lesions, and therefore also possibilities of their application in clinical practice.

Diagnostic accuracy of uterine fluid lactate dehydrogenase isoenzyme activity profile and vaginal ultrasound in detecting endometrial cancer in women with postmenopausal bleeding.

OBJECTIVE: To evaluate the diagnostic accuracy of lactate dehydrogenase (LD) isoenzyme activity profile in uterine fluid and transvaginal ultrasound (TVS) in detection of endometrial cancer in women with postmenopausal bleeding. STUDY DESIGN: Prospective controlled clinical study. METHODS: One hundred twenty postmenopausal women with one or more episode of vaginal bleeding were studied. Endometrial thickness was classified as >5 mm or <5 mm on TVS. LD isoenzyme activity profile was described as normal or abnormal. LD isoenzyme profile was subsequently related to histopathological diagnosis. RESULTS: Endometrial carcinoma was found in 10 out of 120 patients (8.3%). Sixty-seven patients (56%) were found to have endometrium >5-mm thickness on TVS. LD isoenzyme activity profile was abnormal in 22 (18.3%) cases. Histopathological diagnosis in these cases revealed 10 endometrial cancer and 12 benign endometrium. LD isoenzyme activity profile has 100% sensitivity, 90.1% specificity, 45.4% positive predictive value, 100% negative predictive value, and 88.6% accuracy. CONCLUSION: LD isoenzyme profile of uterine fluid could be added as a marker for endometrial cancer in postmenopausal women with vaginal bleeding and endometrial thickness >5 mm on transvaginal ultrasound. Uterine fluid sampling is easy, highly reliable, minimally invasive, with high patient compliance, and can be performed as an office procedure. Furthermore, this method is insensitive to endometrial thickness, amount of sample, sampling device, and dilution.

Cyclooxygenase-2 expression in the endometrium and its relationship to bleeding in users of continuous oral contraceptives.

OBJECTIVE: The objective of the present study was to investigate whether or not the presence of irregular bleeding during use of oral contraceptives (OC) is associated with increased cyclooxygenase-2 (COX-2) expression. PATIENTS AND METHODS: An observational study was carried out in 26 patients who were using gestodene 75 microg/ethinylestradiol 30 microg prior to endometrial resection. The patients were divided into two groups: those with amenorrhea (n = 14) and those who had irregular bleeding (n = 12). The resected endometrium was immunostained for COX-2, Bcl-2 and Ki-67 expression. Routine pathology was carried out using standard hematoxylin-eosin staining. RESULTS: Irregular bleeding during OC use was associated with strong COX-2 expression in both glandular and superficial epithelium. There were also more patients in this group with irregular endometrial maturation and higher Ki-67 values. Bcl-2 expression, on the other hand, was not affected by the presence of uterine bleeding. CONCLUSION: The presence of irregular bleeding during OC use is associated with strong COX-2 expression in the endometrium, thereby suggesting a pivotal role of prostaglandins in this process.

[LD isoenzyme in uterine aspirates a potential marker of endometrial cancer. Investigation of postmenopausal bleeding can be simpler and cheaper]. / LD-isoenzym i uterusaspirat potentiell markör för endometriecancer. Utredning av postmenopausal blödning kan bli enklare och billigare.

In diagnosing endometrial carcinoma in women with postmenopausal bleeding analysis of lactate dehydrogenase, LD, isoenzymes in uterine aspirates appeared to have 100 percent sensitivity and negative predictive value combined with high specificity and positive predictive value. Determination of the LD-profile is suggested as a marker for endometrial carcinoma in women with postmenopausal bleeding. Transvaginal ultrasonography might be combined with determination of the LD-isoenzyme profile to secure the diagnosis of endometrial malignancy in order to minimize the use of other more invasive diagnostic methods.

Diagnosis of endometrial cancer in patients with postmenopausal bleeding by analysis of the lactate dehydrogenase isoenzyme activity profile in uterine fluid.

OBJECTIVES: We have previously shown that high activity of lactate dehydrogenase (LD) isoenzymes 4 and 5 in terine aspirates is a marker for endometrial carcinoma. The purpose of this study was to identify an abnormal activity profile of LD2-5 using LD1 as an internal standard, thereby being able to dispense with measurement of the total LD activity. The profile was subsequently tested for diagnostic power in clinical settings. METHODS: We used data from 11 cases of endometrial cancer. Each isoenzyme was estimated relative to the activity of LD1 (LD1 = 1.0). Based on the lowest level found for each of LD2-5 in the 11 cases, the cut-off levels for an "abnormal profile" were identified. The abnormal profile was subsequently tested for diagnostic power in a group of postmenopausal women at risk for endometrial cancer, that is, they had experienced vaginal bleeding (n = 100). A second group of asymptomatic postmenopausal women (n = 366) had endouterine aspiration performed as part of a regular gynecologic check up to evaluate the prevalence of an abnormal LD isoenzyme profile. RESULTS: In the group of postmenopausal women who had experienced vaginal bleeding, abnormal profile was found in 27 cases: 14 with adenocarcinoma and 13 with benign histology. All cases with normal profile had benign histology. Thus, sensitivity as well as the negative predictive value was 100%. In the group of asymptomatic women, abnormal LD isoenzyme profile was found in 15 cases (4.1%). All had benign histology. CONCLUSIONS: The LD isoenzyme profile detects endometrial malignancy with high accuracy, and equally important, a normal profile excludes malignancy. The profile, which uses relative rather than absolute activity levels, based on LD1 as an internal standard, has the great advantage of being independent of both the dilution factor and the aspiration technique. Larger studies comparing the LD isoenzyme activity profile with ultrasonographic evaluation and biopsy histology are needed.

Mifepristone-induced vaginal bleeding is associated with increased immunostaining for cyclooxygenase-2 and decrease in prostaglandin dehydrogenase in luteal phase endometrium.

The mechanism of mifepristone-induced vaginal bleeding and endometrial shedding was investigated in 13 women who took 200 mg mifepristone in the midluteal phase on d 8 after the onset of the urinary LH surge (LH+8). Endometrial biopsies were collected, 6-24 h after mifepristone (group 1, n = 7) or 36-48 h after mifepristone (group 2, n = 6), and compared with those from a control group in the midluteal phase (n = 7). All women reported vaginal bleeding commencing 36-48 h after taking mifepristone. Treatment with mifepristone significantly reduced serum progesterone levels in all women, when compared with the controls (13.2 nM vs. 34.8 nM, P = 0.001). After mifepristone, a significant increase in cyclooxygenase-2 immunoreactivity was apparent at 36-48 h (P = 0.0018), whereas prostaglandin 15 dehydrogenase enzyme-positive immunostaining declined, to be virtually absent by 36-48 h in both glands and in stroma (P < 0.05). There was no change in intensity or distribution of staining for steroid receptors after mifepristone. The changes in immunostaining for cyclooxygenase-2 and prostaglandin 15 dehydrogenase strongly support the hypothesis that an increase in the local concentration of prostaglandins in the endometrium is involved in the mechanism of bleeding induced by mifepristone in the luteal phase.

Endometrial bleeding in hormone replacement therapy users: preliminary findings regarding the role of matrix metalloproteinase 9 (MMP-9) and tissue inhibitors of MMPs.

OBJECTIVE: To establish the effect of hormone replacement therapy (HRT) on the expression of matrix metalloproteinase 9 (MMP-9) and the tissue inhibitor of MMPs, TIMP-1, in the endometrium of postmenopausal and perimenopausal women. DESIGN: Prospective observational study. SETTING: United Kingdom teaching hospital. PATIENT(S): Thirty-one perimenopausal and postmenopausal HRT recipients, with a control group of eight postmenopausal women not undergoing HRT. INTERVENTION(S): Prospective record of bleeding patterns and endometrial biopsy. MAIN OUTCOME MEASURE(S): Endometrial histology, bleeding patterns, MMP-9, and TIMP-1 expression. RESULT(S): MMP-9 and TIMP-1 are expressed in benign postmenopausal endometrium. Expression of both molecules is reduced in HRT recipients compared with non-HRT recipients. CONCLUSION(S): Exposure to HRT appears to alter endometrial expression of MMP-9 and TIMP-1 and also the local balance between these molecules. This alteration may promote breakdown of the endometrial extracellular matrix and blood vessels and hence bleeding.

The role of matrix metalloproteinases and leukocytes in abnormal uterine bleeding associated with progestin-only contraceptives.

Progestin-only contraceptives are associated with menstrual bleeding disturbances; a major reason why these agents are discontinued. The pathogenesis of abnormal uterine bleeding associated with progestin-only contraceptives remains ill-defined. Matrix metalloproteinases (MMPs) and leukocytes are postulated to be involved in the process of normal menstruation. Immunolocalization of MMPs and leukocytes in (Norplant), and injectable depot medroxyprogesendometrium from women using the progestinterone acetate (DMPA), are widely used, safe and only contraceptives, Norplant or depot medroxyprogesterone acetate (DMPA) compared with normal controls, revealed foci of positive MMP-1 and -3 immunostaining in stromal cells and adjacent extracellular matrix, the presence of MMP-9 in various subtypes of leukocytes and alterations in mast cell phenotype. In women using progestin-only contraceptives, extent of endometrial MMP, neutrophil and eosinophil immunolocalization and the mast cell activation state was similar to or greater than that observed in perimenstrual control women. However, differences in MMP immunostaining were observed in endometrial samples from women using different progestin-only contraceptive agents; in particular, significantly higher MMP-1 immunostaining was observed associated with the use of Norplant compared with DMPA. No correlation was observed with the number of bleeding days recorded. These results suggest that MMP and leukocytes may be involved in endometrial breakdown in women using progestin-only contraceptives.