Platelet Count After Chemotherapy is a Predictor for Outcome for Ovarian Cancer Patients.

Elevated platelet count occasionally is associated with gynecologic malignancies. We investigated the level of platelet count in 450 patients with gynecologic tumors. Ovarian cancer patients have increased platelet count associated with the course of treatment and disease progression. In multivariate analysis, the decrease of platelet count less than 25% after chemotherapy was an unfavorable prognostic factor for PFS (HR, 1.948; 95% CI, 1.083-3.505; p = 0.026) and overall survival (HR, 2.093; 95% CI, 1.022-4.287; p = 0.043). An insufficient decrease of the platelet count increased the risk of recurrence. Thus platelet count could be used for monitoring the disease progression and to predict treatment response.

mtDNA(4977) deletion is not a common feature in patients with premature ovarian failure and primary infertility.

The aim of the current study was to investigate the incidence of mtDNA(4977) deletion in peripheral blood leukocytes of patients diagnosed with premature ovarian failure (POF) and primary infertility. The study group consisted of 17 patients with POF, 32 women with primary infertility, and 31 fertile women with the prevalence of the mtDNA(4977) deletion using the reverse transcription-polymerase chain reaction (RT-PCR) based technology. None of the patients affected by POF revealed mtDNA(4977) deletion. This deletion was detected only in one 26-year-old infertile patient. No significant difference in relation to mtDNA(4977) deletion was reported between the groups investigated (p > 0.05). In conclusion, mtDNA(4977) deletion is not a common finding in peripheral blood leukocytes of women affected by POF and primary infertility. The occurrence of mtDNA(4977) deletion in women between 20 and 39 years of age may not increase with increasing patients' age, independently of their fertility status.

hMLH1 promoter hypermethylation and MSI status in human endometrial carcinomas with and without metastases.

We investigated the methylation status of mismatch repair gene hMLH1 in 80 primary human endometrial carcinomas (ECs) and in 30 metastatic lesions. It was correlated to the expression of hMLH1 protein, microsatellite instability (MSI) of ECs and to the well-known clinico-pathological variables of cancer. The hMLH1 promoter methylation was detected in 24 out of 64 (37.5 %) primary ECs but only in one out of 18 (5.6 %) metastatic lesions investigated. Promoter hMLH1 hypermethylation was found more often in early stage ECs and was associated with a decrease of hMLH1 protein expression immunohistochemically. An inverse relationship between hMLH1 expression and clinical stage of the disease was found (p = 0.048). Interestingly, there was a significant correlation between MSI and hMLH1 protein expression level (p = 0.042). MSI phenotype was found more often in EC metastases compared to the primary tumors (66.7 % vs 29.3 %; p = 0.039). However, neither hMLH1 promoter hypermethylation nor MSI was independent predictive factors for patient's outcome. Using an in vitro model we showed that hMLH1 methylation is reversible. These data showed that hMLH1 methylation with a consequent protein decrease occurred early during EC tumorigenesis and may cause a MSI phenotype, which occurs relatively late. MSI may be an important mechanism supporting further the tumor progression. These findings may have importance for the specific chemosensitization of the primary tumors/metastases and can improve our understanding of endometrial carcinogenesis in humans.

Ovarian endometrioma in an 11-year-old girl before menarche: a case study with literature review.

BACKGROUND: To date, a limited number of endometriosis cases occurring before or around the time of menarche have been documented. CASE: An 11-year-old adolescent underwent an emergency operation for left ovarian endometrioma. Her menarche occurred spontaneously 6 months after surgery. RESULTS AND CONCLUSIONS: We discuss the above mentioned case and consider data published in the literature. Endometrioma should be considered even in premenarcheal girls with ovarian cysts, regardless of their size.

Synchronous adult-type granulosa cell tumor of the ovary with ovarian fibroma: a case report.

The coexistence of two different types of sex-cord stromal tumors, with various clinical symptoms, is extremely rare. We report a case of a 73-year-old woman showing coexistence of adult-type granulosa cell tumor in one ovary with ovarian fibroma in the other. Simultaneously, she was affected by Meigs' syndrome and simple endometrial hyperplasia without nuclear atypia. The different clinical symptoms of the disease according to the available literature are discussed.

Giant uterine leiomyomas causing bilateral hydronephrosis coexisting with endometrial cancer in polyp: a case study.

Multiple uterine leiomyomas are present in a large population of women and may cause several uncommon clinical symptoms, including disseminated vein thrombosis and hydronephrosis. We report a case of giant uterine leiomyomas causing bilateral hydronephrosis coexisting with endometrial cancer (EC) deriving from a uterine polyp. A 50-year-old woman was admitted due to bilateral hydronephrosis caused by monstrous abdominal tumor to the IInd Department of Gynecology, Lublin Medical University, Lublin, Poland. A bilateral double-J catheter was inserted. Pelvic examination revealed a huge, rough tumor, originating from the uterus. Total abdominal hysterectomy with bilateral salpingo-oophorectomy was performed, and a giant uterus weighing 15.2 kg and measuring 35 x 29 x 18 cm was removed. Histopathological examination revealed multiple uterine leiomyomas with calcification and partial necrosis, and well-differentiated (G1), endometrioid-type EC (Stage IA) concomitant with atypical endometrial hyperplasia, deriving from a uterine polyp. The postoperative recovery was without complications, and the patient was discharged on postoperative day 10. In conclusion, giant uterine leiomyoma may incidentally compress the urinary tract organs, causing hydronephrosis.

The significance of C3435T point mutation of the MDR1 gene in endometrial cancer.

The P-glycoprotein (P-gp) plays an important role in carcinogen distribution and is connected with cell differentiation and apoptotic processes leading to carcinogenesis. Interindividual differences in P-gp activity could modulate susceptibility to cancer development. The MDR1 gene, coding for P-gp, is highly polymorphic and some mutations modulate P-gp activity. Recently, association between the MDR1 C3435T polymorphism and the cancer susceptibility was shown. We have hypothesized that MDR1 polymorphism could influence endometrial cancer susceptibility. We have matched 198 women with endometrial cancer and 198 controls. An additional group of 488 healthy volunteers was investigated. The MDR1 C3435T polymorphism was tested by LightCycler assay. The distribution of MDR1 3435 genotypes was significantly different between cases and controls (P = 0.006). Genotypes containing at least one 3435T allele were statistically significant more frequent in the endometrial cancer group (86.8% vs 75.2%, OR 2.18, P = 0.004). Our observation suggests that MDR1 C3435T polymorphism is correlated with endometrial cancer susceptibility.

K-ras gene point mutations and p21ras immunostaining in human ovarian tumors.

It is well recognized that genetic alterations within oncogenes, tumor suppressor genes, DNA mismatch repair and excision repair genes contribute to tumorigenesis within the human ovary. This study was undertaken to screen for the existence of K-ras gene point mutations in paraffin-embedded slides randomly selected from benign and malignant ovarian tumors applying the PCR-RFLP technique. Expression of p21ras was also assessed in 30 primary ovarian adenocarcinomas immunohistochemically. K-ras codon 12 point mutations occurred in nine of 40 (22.5%) cases. They were not identified in two benign mucinous cystadenomas, but in one out of two (50%) mucinous tumors of LMP (low malignant potential), in five out of 30 (17%) ovarian adenocarcinomas, and in one case of adenocarcinoma metastatic to the ovary. K-ras activation was also detected in one out of four (25%) sex cord-stromal cell tumors (folliculoma), and in one dysgerminoma. None of these tumors exhibited K-ras codon 13 point mutations. Gene alterations were more frequently found in mucinous than in non-mucinous (30% vs 10%) tumors, although the difference did not reach significance (p > 0.05). The frequency of K-ras point mutations was correlated neither with clinical nor with pathological variables of cancer. Cytoplasmic p21ras was expressed in all adenocarcinomas negative for K-ras point mutations, whereas one of five (20%) K-ras-positive tumors exhibited lack of immunoreactivity. In conclusion, these findings confirm the role of K-ras activation in mucinous ovarian tumors. p21ras expression is not necessarily associated with K-ras gene alterations in human ovarian adenocarcinomas.

"Low-risk" and "high-risk" HPV-infection and K-ras gene point mutations in human cervical cancer: a study of 31 cases.

To analyze the coexistence of human papilloma virus (HPV) infection and K-ras gene activation in cervical neoplasia, we investigated 31 (seven pre-invasive and 24 invasive) cervical carcinomas for "low-risk" (types 6 and 11) and "high-risk" (types 16 and 18) HPVs and K-ras point mutations using PCR-based technology. "Low-risk" HPVs were not detected in the group investigated; however, 20 of 31 (64%) cases were HPV 16 positive, while HPV 18 was found in only three (9.7%) samples (HPV 6/11 v. HPV 16/18, p < 0.0001; HPV 16 v. HPV 18, p < 0.0001; Fisher's exact test). There was a K-ras codon 12 point mutation in two of 31 (6.4%) neoplasms, with none of the cases showing a K-ras codon 13 point mutation. Two moderately differentiated squamous carcinomas showed K-ras exon 2 gene alterations. Interestingly, none of the pre-invasive cervical carcinomas displayed K-ras gene point mutations. The mean patient age did not differ significantly in the number of HPV-positive and -negative cases. A coexistence of "high-risk" human papillomavirus DNA with K-ras gene alterations was observed in three of 31 (9.7%) neoplasms (one IIA and two IB moderately differentiated cervical carcinomas). Our results suggest that "high-risk" HPVs coexist with K-ras gene alterations in a subset of moderately differentiated carcinomas of the cervix uteri.

Immunohistochemical analysis of MIB-1 proliferative activity in human endometrial cancer. Correlation with clinicopathological parameters, patient outcome, retinoblastoma immunoreactivity and K-ras codon 12 point mutations.

To test the prognostic utility of MIB-1 in human endometrial neoplasias, the proliferative activities of fifty-two endometrial carcinomas obtained from Polish women were assessed. We also investigated the relationship between the MIB-1 Proliferative Index and the well-known clinicopathological features of cancer (clinical stage, histological type, histological grade, depth of myometrial invasion), patient's age, overall survival, retinoblastoma immunostaining and K-ras codon 12 point mutations. The mean MIB-1 Proliferation Index was 43.8%, with a median of 36.0%. Due to the great intratumour heterogeneity of the immunoreaction, the Index ranged from 0% to 98%. A significant relationship was noted between MIB-1 expression and histological grading (p = 0.0004) and myometrial invasion of cancer (p = 0.01). Multivariate Cox regression demonstrated that the clinical stage was the only independent prognostic factor during follow-up (p = 0.025). There was a tendency towards a poorer outcome for women with a Proliferative Index of > 31% than for patients whose Index was < or = 31%; the difference, however, did not reach significance (p = 0.25; log-rank test). Interestingly, uterine cancers lacking retinoblastoma protein expression had a mean MIB-1 Proliferation Index that was nearly twice as high as in those neoplasias that stained positively for retinoblastoma (70.33% and 42.14%, respectively; p = 0.09; Mann-Whitney-U test). There were no significant differences between K-ras codon 12 point mutation-positive and -negative endometrial carcinomas regarding the proliferative activity of the cancer (mean Indexes 47.6% and 43.8%, respectively; p = 0.66, Mann-Whitney-U test). Our data support the view that MIB-1 proliferative activity was significantly increased with a decrease of the histological grading and with the myometrial invasion of human endometrial cancer.

K-ras exon 2 point mutations in human endometrial cancer.

In the present study, we screened for the K-ras exon 2 point mutations in a group of 87 gynecological neoplasms (82 endometrial carcinomas, four carcinomas of the uterine cervix and one uterine carcinosarcoma) using the non-isotopic PCR-SSCP-direct sequencing techniques. Direct sequencing analysis revealed CAA-->CAC (Gln-->His) K-ras codon 61 point mutations in two (2.4%) of the 82 endometrial carcinomas mentioned above. These two cases were endometrial endometrioid carcinomas at an early clinical stage of disease (stage IB and IC due to FIGO). Those endometrial carcinomas that showed K-ras exon 2 point mutations revealed a strong positivity for heterogeneous nuclear retinoblastoma protein staining; none of these, however, have had the K-ras codon 12 point mutation. In addition, there were no K-ras gene point mutations in three endometrial carcinomas lacking the Rb protein immunohistochemically. None of the cervical carcinomas tested had K-ras gene point mutations, whereas one carcinosarcoma harbored K-ras codon 61 point mutation (CAA-->CAC). In conclusion, our data support the view that K-ras exon 2 point mutations are rare events in human endometrial cancer. Rb and K-ras gene abnormalities may occur independently of each other during endometrial carcinogenesis in humans.

[Assessment of the PCNA and P53 proteins expression in the proliferative, hyperplastic and neoplastic human endometrium]. / Immunohistochemiczna ocena ekspresji PCNA oraz onkoproteiny p53 we wzrostowym, rozrostowym i nowotworowym endometrium u kobiet.

In the current study, PCNA and p53 proteins were immunohistochemically studied in the proliferative (n = 5), hyperplastic (n = 4) and neoplastic (n = 20) human endometrium. PCNA immunostaining was noted in 2 out of 5 (40%) proliferative, 4 out of 4 (100%) hyperplastic, and in 18 out of 20 (90%) neoplastic slides. Concomitant PCNA and p53 expression was reported in 12 out of 20 (60%) malignant tumors. All non-endometrioid neoplasms were PCNA-positive, suggested this proliferative marker is commonly expressed in the unfavorable histological types of endometrial cancer.

Total sialic acid content in endometrial cancer tissue in relation to normal and hyperplastic human endometrium.

The aim of this study was to measure the total sialic acid (TSA) content in endometrial cancer tissue and to assess its relationship to clinicopathologic features of the malignancy. Tissue TSA content was measured in 42 women with endometrial cancer, 14 women with endometrial hyperplasia, and 45 women with normal endometrium in the proliferative phase (n = 16) and secretory phase (n = 29) of the menstrual cycle using the Warren procedure. The mean TSA content in endometrial cancer (2.16 micromol/gm) was significantly higher in comparison to normal endometrium in both proliferative (1.23 micromol/gm) and secretory (1.51 micromol/gm) phases. TSA content in the hyperplastic endometrium (1.56 micromol/gm) was higher as compared to the normal endometrium, but the differences were not statistically significant. An increased TSA content in the neoplastic endometrium in relation to the normal endometrium supports the view that the development of endometrial cancer is associated with the increasing content of sialic acid in the tumor tissue.

[Prognostic value of serum MMP-2 level in uterine cancer affected women]. / Prognostyczne znaczenie poziomu MMP-2 w surowicy krwi w przypadku gruczolakoraka blony sluzowej macicy u kobiet.

OBJECTIVES: Matrix metalloproteinase-2 (MMP-2) which can degrade type IV collagen is implicated in cancer invasion and metastasis. The aim of the study was to evaluate the serum MMP-2 level in patients with endometrial carcinoma (EC) and to compare the level with histological grade G1 to G3 in relation to clinical staging I degree-IV degrees as well as myometrial invasion (M. < 1/2, M. > 1/2). MATERIALS AND METHODS: The study group consisted of 30 patients with EC. MMP-2 serum levels were measured with specific one-step sandwich enzyme immunoassay (Amersham Life Science). Statistical analysis was performed by Mann-Whitney test and p value < 0.05 was considered as statistically significant. RESULTS: The statistically elevated enzyme level was observed in patients presenting II degrees-IV degrees clinical stage of EC in comparison to women with I degree stage of this carcinoma. There were no significant correlations between MMP-2 serum levels and grades of histological differentiation as well as depth of myometrium invasion. CONCLUSIONS: MMP-2 serum level is statistically higher in clinically advanced stages of EC. This enzyme seems to be useful in monitoring of therapeutical efficacy or screening for the recurrences of the disease.

Epidermal growth factor receptor immunostaining and epidermal growth factor receptor-tyrosine kinase activity in proliferative and neoplastic human endometrium.

Epidermal growth factor receptor (EGFR) expression and EGFR-tyrosine kinase (EGFR-TK) activity were measured in proliferative (n = 12) and neoplastic (n = 31) human endometrium. Immunoreactivity of EGFR was related to clinicopathological features, estrogen receptor (ER) and progesterone receptor (PR) status, and patient outcome. All proliferative and 27 neoplastic specimens expressed the EGFR. Expression of the EGFR was higher in proliferative endometrium than in endometrial cancer (p < 0.0001). ER immunostaining was observed in 19 of the endometrial carcinomas, while PR expression was demonstrated in only 12 neoplastic specimens. EGFR expression was not related to the ER/PR immunostaining in endometrial carcinomas. Clinicopathological features (age, stage, histological type, grade or depth of invasion) and clinical outcome were unrelated to EGFR immunoreactivity. EGFR-TK activity was detected in 29 of 31 endometrial neoplasms with a 9 times higher mean activity in neoplastic than in proliferative endometrial specimens. There was no relationship between the EGFR-TK activity and EGFR immunostaining in human neoplastic endometrium (p = 0.77). A trend towards a poorer outcome of patients with the EGFR-TK activity above 40 pmol/min/mg was observed, but was not statistically significant. These results support the view that the EGFR expression is downregulated in endometrial carcinomas compared to proliferative endometrial specimens.

Detection of human papillomavirus types 16 and 18 in human neoplastic endometrium: lack of correlation with established prognostic factors.

HPV (types 16 and 18) DNA sequences are present in the majority of precancerous and cancerous lesions of the human uterine cervix. However, data concerning the involvement of HPVs infection in the pathogenesis of endometrial cancer are controversial. In the current study we investigated the frequency of the HPV types 16 and 18, detected by PCR amplification using the type 16- and 18-specific primers within the E7 Open Reading Frame (ORF) sequence, in 54 human endometrial carcinomas obtained from women of Polish origin. Moreover, we assessed the possible association of the HPV with the clinicopathological features of the cancer, patients' outcome as well as with the K-ras codon 12 gene point mutations. HPV type 16 was present in eleven out of 54 (20%) endometrial tumors, while HPV type 18 was detected only in three out of 54 (4%) neoplasms analyzed. HPV infection was not related either to the patients' age (r=0.11; p=0.428, Spearman correlation test) or to the clinicopathological parameters and patients' prognosis. A higher incidence of HPV 16/18 was detected in well (G1) differentiated than in moderately (G2) and poorly (G3) differentiated endometrial adenocarcinomas, but the difference was not statistically significant. Moreover, none of HPV-positive endometrial carcinomas harbored K-ras codon 12 gene point mutations. Our results suggest that some of the endometrial carcinomas are associated with HPV infection but the presence of the human papillamovirus types 16/18 is not related to the clinicopathological or prognostical features of the neoplasm.

Detection of CFTR gene mutations in patients suffering from chronic bronchitis.

BACKGROUND: The purpose of the study was to examine cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in patients suffering from chronic bronchitis. METHODS: Thirty-two patients admitted to the Department of Pulmonology, Lublin School of Medicine, Lublin, Poland between 1995 and 1996 due to chronic bronchitis were included in the study. Patients were analyzed for the eight most common mutations of the CFTR gene (DeltaF508, G542X, N1303K, 1717-1(GoA)), W1282X, G551D, R553X, and DeltaI507 by the reverse-hybridization method. RESULTS: CFTR gene mutations were found in five of 32 (16%) patients, all within the DeltaF508 region of the CFTR gene. All positive samples were obtained from patients heterozygous for the DeltaF508 mutation. The presence of the DeltaF508 mutation was considered statistically significant when our study group was compared to the study of Poland's general population (p <0.05 Fisher's exact test). CONCLUSION: Our results suggest there is an increased presence of the DeltaF508 point mutation of the CFTR gene in Polish patients suffering from chronic bronchitis.

p53 protein detection by the western blotting technique in normal and neoplastic specimens of human endometrium.

The aim of the study was to investigate p53 protein expression by the Western blotting technique (estimated by integrated optical density - IOD) in normal (n = 13) and neoplastic (n = 40) human endometrial tissues as well as in a case of uterine carcinosarcoma and in a specimen of the botryoid sarcoma of the uterine cervix. p53 protein levels were correlated with patients' age as well as with conventionally used clinicopathological features of the endometrial neoplasm. A statistically significant difference was noted in p53 levels in the nuclear, but not in the cytoplasmatic, fraction between the normal endometria and endometrial cancer tissues (P < 0.0001). In the neoplastic endometria, nuclear p53 protein expression was higher than in cytoplasmatic fraction, and the difference was significant (P < 0.05). Higher nuclear p53 protein levels correlated with advanced histological grading of endometrioid endometrial carcinomas, but no relationship was noted between p53 protein expression and patients' age, clinical stage, histological type or depth of myometrial invasion. A case of uterine carcinosarcoma and a specimen of a botryoid sarcoma of the uterine cervix expressed nuclear p53 oncoprotein (57 IOD and 89 IOD, respectively). In conclusion, we found a statistically higher nuclear p53 levels in malignant as compared to normal human endometrial specimens by the Western blotting technique. Although there were no significant differences between p53 expression and clinicopathological features of the neoplasm (except poor histological grading), further studies are necessary to evaluate the influence of p53 nuclear/cytoplasmatic levels on the clinical outcome of Polish patients suffering from endometrial cancer.

RB protein expression in human endometrial carcinomas--an immunohistochemical study.

The aim of the current study was to investigate the immunohistochemical expression of the retinoblastoma protein (pRB) in formalin-fixed, paraffin-embedded specimens obtained from 62 patients suffering from endometrial cancer. The avidin-biotin-peroxidase detection system with microwave pretreatment and the mouse anti-human NCL-RB1 monoclonal antibody were used. Heterogeneous nuclear immunostaining for the pRB was generally observed in the glandular cells in 59 out of 62 (95%) endometrial carcinomas, while stromal components were unreactive. In one case of stage Ic endometrioid adenocarcinoma, a small percentage of glandular cells (5%) stained positively with the anti-RB antibody, while two other tumors (stage IIa adenosquamous carcinoma and stage IIIa endometrioid adenocarcinoma) were pRB negative. In the cases with concomitant hyperplastic and neoplastic endometrial lesions, pRB immunoreaction was heterogeneous in the hyperplastic endometrial cells and in the adjacent neoplastic endometrium. Moreover, eight cases of endometrial carcinoma harboring K-ras codon 12 gene point mutation overexpressed pRB (more than 80% of glandular endometrial cells were positive) immunohistochemically, while none of three pRB negative slides had a K-ras gene alteration. Our data support the view that the pRB is expressed in most of the human endometrial neoplasms, but the lack of pRB immunoreactivity may correspond with the retinoblastoma gene rearrangements in a subset of advanced endometrial carcinomas.

Analysis of p53 and K-ras genes and their proteins in a sarcoma botryoides of the uterine cervix.

Several data indicate that the activation of oncogenes and growth factors as well as inactivation of the tumor suppressor genes are implicated in the development of human neoplasms, including sarcomas. In the present study we described a case of the extremely rare, but highly malignant neoplasm of the female genital tract known as sarcoma botryoides of the uterine cervix and assessed, using molecular and an immunohistochemical analysis, p53 and K-ras alterations in the tumor. A point mutation in exon 6 of the p53 tumor suppressor gene was found but no K-ras gene point mutations at codons 12, 13 and 61 were detected using molecular analysis. p53 protein was overexpressed in more than half of the neoplastic cells, however, ras p21 protein expression was not immunohistochemically detected. Our data indicate that p53, but not K-ras gene alterations may play a role in the development and progression of sarcoma botryoides of the uterine cervix.