TLR4 signaling induced by lipopolysaccharide or paclitaxel regulates tumor survival and chemoresistance in ovarian cancer.

Toll-like receptors (TLRs) expressed on immune cells trigger inflammatory responses. TLRs are also expressed on ovarian cancer (OvCa) cells, but the consequences of signaling by the TLR4/MyD88 pathway in these cells are unclear. Here, TLR4 and MyD88 expression in OvCa tissues (n=20) and cell lines (OVCAR3, SKOV3, AD10, A2780 and CP70) was evaluated by reverse transcriptase-PCR, western blots and immunohistochemistry. Cell growth, apoptosis, nuclear factor-kappaB (NF-kappaB) translocation, IRAK4 and TRIF expression and cJun phosphorylation were measured following tumor cell exposure to the TLR4 ligands, lipopolysaccharide (LPS) or paclitaxel (PTX). Culture supernatants were tested for cytokine levels. TLR4 was expressed in all tumors, tumor cell lines and normal epithelium. MyD88 was detectable in tumor tissues and in 3/5 OvCa lines but not in normal cells. In MyD88(+) SCOV3 cells, LPS or PTX binding to TLR4 induced IRAK4 activation and cJun phosphorylation, activated the NF-kappaB pathway and promoted interleukin (IL)-8, IL-6, vascular endothelial growth factor and monocyte chemotactic protein-1 production and resistance to drug-induced apoptosis. Silencing of TLR4 in SCOV3 cells with small interference RNA resulted in phosphorylated-cJun (p-cJun) downregulation and a loss of PTX resistance. In PTX-sensitive, MyD88(neg) A2780 cells, TLR4 stimulation upregulated TRIF, and TLR4 silencing eliminated this effect. Thus, TLR4/MyD88 signaling supports OvCa progression and chemoresistance, promoting immune escape.

5-aminolevulinic acid-mediated photodynamic therapy of human endometriotic primary epithelial cells.

OBJECTIVE: Despite progress in medicine, appropriate diagnosis and treatment of endometriosis poses a serious problem. For this reason, in in-vitro experiments were performed on a potential method of employing photodynamic therapy (PDT) of endometriosis using 5-aminolevulinic acid (ALA). BACKGROUND DATA: The exogenous application of ALA induces the accumulation of protoporphyrin IX, a natural and effective photosensitizer used in photodynamic diagnosis and therapy. MATERIALS AND METHODS: To this end primary epithelial cells were isolated from endometriotic foci, preincubated with various ALA concentrations, and then exposed to light energy (a bulb or laser) for 8 min; 24 and 48 h later cell lesions were evaluated using fluorescent staining. RESULTS: When bulb illumination was used, after 48 h cells were found that had disturbed chromatin concentration and fragmentation. Illumination with a laser beam resulted in strong induction of apoptosis 24 h post-exposure. With both types of illumination the number of necrotic cells was insignificant. Staining with rhodamine 123 demonstrated the presence within the endometriotic foci of epithelial cells that were resistant to ALA-induced photodynamic therapy. CONCLUSION: The effects of PDT on primary epithelial endometriotic cells may prove useful in designing a phototherapeutic procedure for the detection and treatment of endometriosis.

Coexpression of human chorionic gonadotropin beta subunit and its receptor in nontrophoblastic gynecological cancer.

A considerable number of biochemical and physiologic studies evaluate the roles of gonadotropins in carcinogenesis. Latest reports show that human chorionic gonadotropin (hCG), and especially its beta subunit, are secreted by a variety of malignant tumors of different origin. However, the mechanism of hCG action and its role in tumor development is not known yet. This study, with the help of reverse transcription-polymerase chain reaction and immunohistochemistry, is an attempt to document the molecular presence of the hCGbeta and luteinizing hormone/hCG receptor (LH/hCGR) in the ovarian, endometrial, and uterine cervix cancer tissues. The LH/hCGR, coexpressed with hCGbeta, may act as a potential mediator of hCG action in nontrophoblastic gynecological cancers.

Photodynamic diagnosis (PDD) using 5-aminolevulinic acid-supplemented cultures of human endometrial epithelial cells.

PURPOSE: The studies were aimed at monitoring 5-aminolevulinic acid (5-ALA)-dependent accumulation of endogenous protoporphyrin IX (PpIX) in epithelial cells originating from normal endometrium or endometriotic foci, as related to steroid treatment. MATERIAL AND METHODS: Epithelial cells were cultured in presence of estradiol-17 beta (E2) and progesterone (P) in concentrations typical for the follicular stage (E2 alone, 220 pg/ml) or the luteal stage (E2 100 pg/ml and P 2 ng/ml) or in presence of progesterone alone (2 ng/ml) for a period of 24, 48 or 72 h. Effect of 5-ALA concentration on the accumulation of PpIX was defined in the cells incubated with 2.0 mmol/l 5-ALA for 2 h. PpIX fluorescence was detected using a confocal microscope. RESULTS: After hormonal stimulation, intensity of PpIX-specific fluorescence was only slightly increased in epithelial cells originating from normal endometrium. Cultures of epithelial cells from endometriosis foci showed higher concentration of PpIX than did the cells originating from normal endometrium. The highest peak of PpIX fluorescence was noted in epithelial endometriotic cells after 48h incubation with progesterone. CONCLUSIONS: The data on PpIX accumulation in epithelial cells in the presence of estradiol-17 beta or progesterone may provide indications as to the menstrual cycle phase(s) in which photodynamic therapy for endometriosis should be performed. It is concluded that hormonal condition of female body must be taken into account for diagnosis and treatment of endometriosis.

Expression of beta-human chorionic gonadotropin in ovarian cancer tissue.

The purpose of the present investigation was to determinate expression of human chorionic gonadotropin gene in ovarian cancer tissue. The study included 15 patients with epithelial ovarian carcinoma. The expression of mRNA hCGbeta was determinated by the RT PCR method and the distribution of the hormone in study tissue was analyzed immunohistochemicaly. In all 15 study specimens of the ovarian carcinoma tissue the active hCGbeta gene was found, whereas noncancerous tissue demonstrated lack of the hormone expression. Thus, the study clearly shows that the expression of hCGbeta is the feature of ovarian cancer tissue.

Human chorionic gonadotropin-beta in endometrium cancer tissue.

PURPOSE: Determination of the correlation between expression of human chorionic gonadotropin mRNA and serum free hCGb immunoreactivity in endometrial cancer tissue. METHODS: The study included 56 patients with endometrial carcinoma Stages IB-III. The expression of mRNA hCGbeta was determined by the RT PCR method in 18 cases of cancerous and precancerous tissues. The serum-free hCGbeta immunoreactivity was analyzed by sequential immunometric assay in all patients. RESULTS: In 15 study specimens of endometrial carcinoma tissue mRNA of hCGbeta was detected. Also in endometrial atypical hyperplasia, expression of hCGbeta was found. Noncancerous tissue demonstrated lack of the hCGbeta transcript. The serum immunoreactivity in the endometrial cancer group was detectable in 86% of cases. There were no significant differences between FIGO stages and grading. CONCLUSION: The results of the present study confirmed the presence of active genes of hCGbeta in endometrial cancer tissue, even in precancerous changes. The serum immunoreactivity of free hCGbeta is a less common feature and is not linked with tumor stage or grade in endometrial cancer patients.

Effect of 5-aminolevulinic acid (ALA) doses and oestrogen/progesterone on protoporphyrin IX (ppIX) accumulation in human endometrial epithelial cells.

Endometriosis represents one of the most frequent causes of restricted fecundity. Despite the progress in medicine, appropriate diagnosis and treatment pose significant problems. The aim of this study was to evaluate ALA-induced PpIX fluorescence of normal endometrial epithelial cells for the diagnosis of endometriosis. PpIX-fluorescence was measured after stimulation with estradiol-17 beta (E2) or with estradiol-17 beta (E2) and progesterone (P) and after incubation with ALA under a confocal microscope. The epithelial cells showed a significantly higher fluorescence of PpIX in the course of 24 and 48h incubation with hormones, than the cells without stimulation. After 72h, a significant decrease in cellular PpIX concentration was noted. The results suggested that E2 and P were required to convert ALA to PpIX in epithelial cells and increased PpIX concentration in a time-dependent fashion.

[Analysis of interleukin-6 serum concentration in trophoblastic tumors prognosis]. / Wartosc kliniczna analizy stezen interleukiny-6 u kobiet chorych na rozrosty i nowotwory trofoblastu.

OBJECTIVES: Our purpose was to estimate the prognosis based on Il-6 concentration in cases of trophoblastic tumors. MATERIALS AND METHODS: The study population was 65 women suffering from hydatiform mola or choriocarcinoma. We divided them into two groups: 30 patients who required only operative management and 35 patients who required operative procedures and additional chemotherapy. The observation period was 6 months. Blood samples were collected every 4 weeks. Concentration of Il-6 was measured in ELISA assay. RESULTS: The serum Il-6 concentration was significantly higher in cases of trophoblastic diseases than in the group of healthy women and higher in patients who required chemotherapy after operation (451.0 +/- 88.5 pg/ml), than in patients treated only surgically (257.1 +/- 77.1 pg/ml). CONCLUSIONS: Patients with hydatiform mola and choriocarcinoma reveal higher concentration of Il-6 than healthy women. It is associated with disease prognosis and allows to determine at the time of establishing a diagnosis, whether a patient can be treated only surgically or requires an additional chemotherapy.

[Chorion carcinoma: failure of treatment]. / Rak kosmówki--niepowodzenie leczenia.

Between 1986-1998 in University Oncology Gynecology Department in Poznan, Poland were treated 23 women with choriocarcinoma. Despite of intensive chemotherapy 3 women were dead. In the report we present the history of this choriocarcinomas making effect to answer why our therapy was ineffective.

[Gestational trophoblastic disease: the epidemiological analysis of 342 cases]. / Rozrosty i nowotwory trofoblastu--analiza epidemiologiczna 342 chorych.

Between 1987-1996 dates were collected to assess frequency and risk factors for gestational trophoblastic disease in a case-control study of 342 women with trophoblastic tumors and 342 pregnant women admitted for deliveries or spontaneous abortion to University Hospitals in Poznan, Poland. Were analyzed the age of women obstetric history, place of live and repeat appearance of hydatidiform mole. The risk of trophoblastic disease increased with increase in maternal age and above third pregnancy. The risk independent of living in town or in the country. The second and more incident of hydatidiform mole was associated with greater risk of malignant sequele. The study of the pregnancy of gestational trophoblastic disease was led in Great Poland in the support on the date from all pathologic centres in this region and public demographic office. The frequently of hydatidiform mole was between 1987-1996 2.32 per 100,000 women, and 0.76 for 1000 live birth (1 HM for 1315 live birth). The frequently of choriocarcinoma was 0.08 per 100,000 women (and 0.38 per 10,000 live birth (1 CHA per 26,315 live birth).

[Glycoforms of alpha-1-acid glycoprotein in trophoblastic tumors prognosis]. / Znaczenie rokownicze glikoform alpha 1-kwasnej glikoproteiny w rozrostach i nowotworach trofoblastu.

OBJECTIVES: Our purpose was to define the pattern of AGP glycolysation and concentration in cases of trophoblastic tumors. MATERIALS AND METHODS: The study population was 65 women suffering from hydatiform mola or choriocarcinoma. We divided them into two groups: 30 patients who required only operative management and 35 patients who required operative procedures and additional chemiotherapy. The observation period was 6 months. Blood samples were collected every 4 weeks. Concentration of AGP was measured in radial immunodyfusion. Glycolysation pattern was defined in Concanavalin A crossed affinoimmunoelectroforetic analysis. RESULTS: The serum AGP concentration was significantly higher in patients treated only surgically (1026.1 +/- 241.0 mg/l) than in patients who required chemotherapy after operation (740.3 +/- 103.0 mg/l). AGP microheterogeneity was represented by 2 to 4 glycoforms. CONCLUSIONS: Patients with hydatiform molar and choriocarcinoma reveal characteristic changes in concentration and microheterogeneity of AGP, which are associated with disease prognosis. They allow to determine at the time of establishing a diagnosis, whether a patient can be treated only surgically or with additional chemotherapy.

[Bowen's disease: one of the types of vulvar intraepithelial neoplasm (VIN) III (four case reports)]. / Choroba bowena--jedna z postaci VIN III (przedstawienie czterech sytuacji klinicznych).

Vulvar intraepithelial neoplasia has been considered a problem of postmenopausal years, but its frequency appears to be increasing among younger women. Bowen's disease is a form of VIN III carcinoma in situ, that may appear clinically as pigmented lesions. This pathology mostly concerns younger women, and is probably connected with human papilloma virus (HPV) infection. The incidence rate is very rare. During the last 15 years only four cases of Bowen's disease have been diagnosed and treated in Division of Gynecological Oncology in Poznan. In this paper we presented symptoms, diagnostic procedures, results of clinical and histopathologic evaluation as well as managements methods. Also the results of treatment and reports on follow-up visits have been discussed.

[Can the DNA-HPV study in vulvar squamous cell carcinoma have a diagnostic value?]. / Czy obecnosc DNA HPV w rakach sromu moze miec wartosc diagnostyczna?

The authors studied 27 vulvar cancer looking for relationship between presents of DNA HPV morphological structure and clinical follow-up. There is suggestion that HPV status may play a profitable role.

[Non-neoplastic epithelial disorders associated with vulvar carcinoma]. / Nablonkowe, nienowotworowe choroby wspólistniejace z rakiem sromu.

The purpose of this paper has been clinical problems associating squamous cell carcinoma of the vulva co-existing with non-neoplastic epithelial disorders. We analyzed 46 women with vulvar carcinoma. Non-neoplastic epithelial disorders we found in 17% cases. In conclusions we suggest the follow-up in patient with non-neoplastic epithelial disorders must be held the same like with vulvar carcinoma.

[Ectopic cervical pregnancy--conservative treatment with methotrexate]. / Ciaza ektopowa szyjkowa--leczona zachowawczo methotrexatem.

Ectopic pregnancy regardless its localization always remains difficult diagnostic and therapeutical problem. Special attention is paid to cervical pregnancy because of its rare incidence and poor prognosis (even in this day and age maternal death rate is 40-45%). In this paper there has been reported the case of 38 years old patient, admitted to Dept of Gynecological Oncology with suspicion of alive cervical pregnancy. Diagnostic procedures have been described as well as positive results of conservative treatment with Methotrexate (under careful monitoring--ultrasound, beta hCG levels). Also there has been presented detailed description of endocervix curettage after fetal death confirmed ultrasonographically. Current conservative therapy methods for cervical pregnancy have been discussed.

[Hysteroscopy in the diagnosis and treatment of residua after evacuation of hydatidiform mole]. / Histeroskopia w diagnostyce i leczeniu zmian wewnatrzmacicznych po usunieciu zasniadu groniastego.

The authors showed the case of 23 years old multipara treated in Department of Reproduction, Institute of Gynecology and Obstetrics, Academy of Medicine, Poznan because of hydatidiform mole using the exhaustion of uteri. After 8 weeks the hysteroscopy was done in order to explain the reasons of elevated serum beta hCG levels. During the hysteroscopy the polypoid lesion was recognized and removed using the electric resectoscope. After this endoscopic operation the normalization of serum beta hCG levels was achieved.