Uterus Tissues Research Articles

The porcine Gpr3 gene: molecular cloning, characterization and expression level in tissues and cumulus–oocyte complexes during in vitro maturation

G protein-coupled receptor 3 (Gpr3) is a member of G protein-coupled receptor rhodopsin family, which is present throughout the follicle within the ovary and functions as a critical factor for the maintenance of meiotic prophase arrest in oocytes by a Gs protein-mediated pathway. In the current paper, attempts were made to clone and characterize a gene encoding Gpr3 from pigs and investigate its expression pattern in tissues and the whole cumulus-oocyte complexes (COCs) in vitro maturation (IVM). Rapid amplification of cDNA ends and RT-PCR gave rise to the full sequence of Gpr3 gene with its length being 2101bp nucleotides, including an open reading frame of 993bp, encoding a 331 amino acid polypeptide with the molecular weight of 35.2kDa. Homology search and sequence multi-alignment demonstrated that the putative porcine Gpr3 protein sequence shared a high identity with other animal Gpr3 orthologs, including several highly conservative motifs and amino acids. Real-time PCR analysis showed that the Gpr3 gene was expressed in tissues of cerebrum, cerebellum, hypothalamus, pituitary, ovary, oviduct, uterus, heart, liver, spleen, lung, kidney, muscle, fat, testis, thymus and granulosa cell, oocyte and COCs at different expression levels. The expression levels of this gene in oocyte, uterus, liver, fat, pituitary and brain were higher than that in other tissues. Interestingly, the mRNA and protein levels of Gpr3 in the whole COCs were down-regulated, and its mRNA expression levels were significantly and negatively correlated with the degrees of cumulus expansion (r=-0.937, P<0.01) during IVM, suggesting its important roles in cumulus expansion and oocyte maturation.

Mystery of bilateral breast masses

Leiomyosarcoma (LMS) is an uncommon malignant tumour of smooth muscle origin. It arises in the gastro intestinal tract, retroperitoneum, urinary bladder, uterus and soft tissue. Peritoneal leiomyosarcomatosis (PL) is defined as a peritoneal dissemination of a primary sarcoma. We present a case of leiomyosarcomatosis with wide spread dissemination including involvement of both breasts.

Implantation Serine Proteinase 1 Exhibits Mixed Substrate Specificity that Silences Signaling via Proteinase-Activated Receptors

Implantation S1 family serine proteinases (ISPs) are tryptases involved in embryo hatching and uterine implantation in the mouse. The two different ISP proteins (ISP1 and ISP2) have been detected in both pre- and post-implantation embryo tissue. To date, native ISP obtained from uterus and blastocyst tissues has been isolated only as an active hetero-dimer that exhibits trypsin-like substrate specificity. We hypothesised that in isolation, ISP1 might have a unique substrate specificity that could relate to its role when expressed alone in individual tissues. Thus, we isolated recombinant ISP1 expressed in Pichia pastoris and evaluated its substrate specificity. Using several chromogenic substrates and serine proteinase inhibitors, we demonstrate that ISP1 exhibits trypsin-like substrate specificity, having a preference for lysine over arginine at the P1 position. Phage display peptide mimetics revealed an expanded but mixed substrate specificity of ISP1, including chymotryptic and elastase activity. Based upon targets observed using phage display, we hypothesised that ISP1 might signal to cells by cleaving and activating proteinase-activated receptors (PARs) and therefore assessed PARs 1, 2 and 4 as potential ISP1 targets. We observed that ISP1 silenced enzyme-triggered PAR signaling by receptor-disarming. This PAR-disarming action of ISP1 may be important for embryo development and implantation.

Age dependency of estrogen responsiveness in the uterus and adipose tissue of aromatase-knockout (ArKO) mice

Aging is often associated with weight gain caused by metabolic changes including an increase of body fat. In this study we assessed the impact of age on estrogen responsiveness in the uterus and adipose tissue (AT) in aromatase-knockout (ArKO) mice. ArKO mice at the age of three or twelve months respectively were treated s.c. with vehicle, E 2 (10 μg/kg BW/d) or genistein (15 mg/kg BW/d) for three days. In the ArKO mouse model we were able to demonstrate that estrogen treatment resulted in an age specific response pattern both on a physiological and molecular level. Assessment of basal gene expression levels revealed significant age dependent differences only for elevated Esr1 levels in the uterus and leptin levels in infrarenal fat as well as lower levels of Pparg in the gonadal fat tissue. Investigating age dependency of estrogen responsiveness we were able to show that the E 2 and genistein resulted in age related pattern of regulation of expression of Esr1 and Lep in infrarenal and gonadal AT as well as the uterine expression of Pgr, Ltf and Pparg. In conclusion, evidence is provided that aging has an impact on the effectiveness of estrogen regulated processes in uterus and AT of ArKO mice. It remains to be elucidated whether or not this is associated with weight gain caused by an increase in body fat mass.

Ex vivo water diffusion tensor properties of the fibroid uterus at 7 T and their relation to tissue morphology

To investigate the water diffusion tensor properties of ex vivo tissue in the fibroid uterus, including the influence of degeneration, and the relevance of the principal eigenvector orientation to the underlying tissue structure. Following hysterectomy, high-resolution structural T(2) -weighted and diffusion tensor magnetic resonance imaging (DT-MRI) were performed on nine uteri at 7 T. Mean diffusivity (MD), fractional anisotropy (FA), and principal eigenvector orientation were measured in myometrium and in myxoid and dense tissue in fibroids. Imaging data and measurements of water diffusion parameters were compared with histopathology findings. The nine uteri yielded 23 fibroids. MD was 50% higher in regions of myxoid degeneration compared to dense fibroid tissue (P = 0.001), while myometrium was intermediate in value (dense fibroid tissue, P = 0.15; myxoid degeneration, P = 0.23). FA was lower in dense fibroid tissue than in myometrium (P = 3 × 10(-5) ), but higher than in myxoid tissue (P = 0.003). Principal eigenvector orientation corresponded qualitatively with that of uterine smooth muscle fibers. The water diffusion tensor measured ex vivo in the fibroid uterus is a sensitive probe of tissue type, myxoid degeneration, and morphology.

Prognostic significance of elongator protein 3 expression in endometrioid adenocarcinoma

Elongator protein 3 (ELP3), the catalytic subunit of the elongator complex of RNA polymerase II, is involved in various functions, including transcriptional elongation, chromatin modification and cytoskeletal regulation. In this study, ELP3 expression was immunohistochemically examined in normal uterus tissue and uterine endometrioid adenocarcinoma tissue. ELP3 was abundantly expressed in both the proliferative and secretory phases of the endometrial cycle. However, ELP3 expression levels varied among cases of endometrioid adenocarcinoma. In patients with endometrioid adenocarcinoma, a low ELP3 expression was correlated to a high T-factor (p=0.036), tumor stage (p=0.001), lymph node metastasis (p<0.001), resistance to chemotherapy (p=0.045), recurrence (p=0.004) and poor prognosis (p=0.003). Univariate and multivariate analyses revealed that a low ELP3 expression was an independent factor for poor prognosis. In conclusion, this is the first study to examine the clinical implications of ELP3 expression in cancer.

Melatonin reduces LH, 17 beta-estradiol and induces differential regulation of sex steroid receptors in reproductive tissues during rat ovulation

BackgroundMelatonin is associated with direct or indirect actions upon female reproductive function. However, its effects on sex hormones and steroid receptors during ovulation are not clearly defined. This study aimed to verify whether exposure to long-term melatonin is able to cause reproductive hormonal disturbances as well as their role on sex steroid receptors in the rat ovary, oviduct and uterus during ovulation.MethodsTwenty-four adult Wistar rats, 60 days old (+/- 250 g) were randomly divided into two groups. Control group (Co): received 0.9% NaCl 0.3 mL + 95% ethanol 0.04 mL as vehicle; Melatonin-treated group (MEL): received vehicle + melatonin [100 μg/100 g BW/day] both intraperitoneally during 60 days. All animals were euthanized by decapitation during the morning estrus at 4 a.m.ResultsMelatonin significantly reduced the plasma levels of LH and 17 beta-estradiol, while urinary 6-sulfatoximelatonin (STM) was increased at the morning estrus. In addition, melatonin promoted differential regulation of the estrogen receptor (ER), progesterone receptor (PR), androgen receptor (AR) and melatonin receptor (MTR) along the reproductive tissues. In ovary, melatonin induced a down-regulation of ER-alpha and PRB levels. Conversely, it was observed that PRA and MT1R were up-regulated. In oviduct, AR and ER-alpha levels were down-regulated, in contrast to high expression of both PRA and PRB. Finally, the ER-beta and PRB levels were down-regulated in uterus tissue and only MT1R was up-regulated.ConclusionsWe suggest that melatonin partially suppress the hypothalamus-pituitary-ovarian axis, in addition, it induces differential regulation of sex steroid receptors in the ovary, oviduct and uterus during ovulation.

CDNA cloning, genomic structure and mRNA expression pattern of porcine type I interferons receptor 2 gene

Type I interferons (IFN) are important mediators of the host defence against viruses through binding to the cell surface receptors, among which the binding to type I IFN receptor 2 (IFNAR2) is the very first step initiating a complex signal transduction cascade. By using RT-PCR and 5' RACE approaches, we obtained porcine IFNAR2 cDNA, the nucleotide identity of its coding region is 57.53%, 67.45%, 74.07% and 74.63% to those of mouse, human, sheep and cattle, respectively; and the deduced protein of which shares 38.18%, 55.29%, 62.01% and 63.39% identity to those of mouse, human, sheep and cattle, respectively. The genomic structure of porcine IFNAR2 gene consists of nine exons and eight introns. Porcine IFNAR2 mRNA expression was detected in all tissues examined, being strong in the spleen, small intestine, cerebrum and uterus tissues and relatively weak in the stomach tissues. As compared with piglets, the expression of IFNAR2 mRNA was significantly higher in both liver and spleen of Laiwu adult pigs (P < 0.01); in Duroc pigs, however, significantly higher IFNAR2 mRNA expression was only found in adult liver (P < 0.05). In Duroc × Landrace × Yorkshire commercial pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV), the expression of IFNAR2 mRNA in lung tissue was significantly down-regulated as compared to uninfected ones (P < 0.05).

The Use of a Prostacyclin Analog, Iloprost, as an Adjunct to Uterus Preservation with Histidine-Tryptophan-Ketoglutarate Solution

Introduction Although assisted reproductive techniques have made most causes of both male and female infertility treatable, uterine factor infertility is not able to therapy. Therefore, transplantation of the uterus has been suggested as a future possible cure. Organ preservation solutions seek to reduce reperfusion injury. Since iloprost is an antioxidant with cytoprotective properties, we investigated its potential positive effects in histidine-tryptophan-ketoglutarate (HTK) solution after 4 or 24 h cold storage period of the rat uterus. Methods We divided 24 female Wistar-albino rats into four groups: Group 1 had the uterus tissue stored in HTK solution at 4 °C for 4h. Group 2, the tissue was stored in HTK solution combined with iloprost (10 −8 M) for 4h at 4 °C. The same procedures were repeated for 24 h for Groups 3 and 4 respectively. Tissue levels of malondialdehyde (MDA) and nitric oxide (NO), as indicators of oxidative stress were determined with histopathological evaluations. Results MDA and NO levels were compared between the group 1 vs 3; and 2 vs 4. No significant difference was observed between the groups. Cold storage for 24 h produced alterations in histological appearences that were mitigated by the addition of iloprost to HTK solution. Conclusion In conclusion, addition of iloprost to HTK solution reversed the histological alterations after 24h-cold storage of the rat uterus.

Abstract 1522: Spheroid formation causes a decrease in ovarian cancer cell growth rate that persists through dissociation and growth as a monolayer

Abstract Ovarian cancer has the highest mortality of all cancers of the female reproductive system. It accounts for three percent of all cancers in women and is the fifth leading cause of cancer-related death among women in the United States. Ovarian cancer cells can metastasize to the uterus, fallopian tubes, and pelvis tissues as well as to other organs. Even with optimal treatment there is a high probability that the cancer will recur, suggesting that current treatments are not successful in eliminating all of the ovarian cancer cells. This may be a result of a protective mechanism of the ovarian cancer cells. Spheroids are large collections of cancer cells that adhere tightly to each other rather than spreading out across an underlying basement membrane. They have been observed in ovarian cancer both in vivo and in ovarian cancer cell lines. Spheroids have been shown to play a role in the metastasis of ovarian cancer cells and have also been shown to be drug resistant. It is our hypothesis that spheroid formation results in epigenetic changes promoting the survival of ovarian cancer cells. Our data suggests that spheroid growth causes the growth rate of ovarian cancer cells to decrease and remain decreased following the transition from spheroids back to monolayer culture. Furthermore, serial passaging of ovarian cancer spheroids results in an increase in resistance to cytotoxic drugs. Since the decrease in growth rate appears to be persistent after spheroid culture it is likely the result of epigenetic modifications that occur during spheroid formation. These epigenetic changes, currently being studied, could become potential drug targets, as new treatments are developed to not only kill ovarian cancer cells but also prevent spheroid formation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1522. doi:10.1158/1538-7445.AM2011-1522

Orbital metastasis of retroperitoneal leiomyosarcoma

Leiomyosarcoma is a relatively rare form of cancer most commonly found in the uterus, stomach, small intestine, retroperitoneum, and vascular tissue. Metastasis is common and surgery with a wide margin of resection is generally considered the most effective management. Leiomyosarcomas, metastatic or primary, of the orbit have rarely been reported. Herein, we report the case of a 51-year-old female with metastatic orbital leiomyosarcoma. The diagnosis was based on her history of leipmyosarcoma, orbital ultrasonography, and computed tomography and confirmed by postoperative histochemistry. She was treated with surgical excision and has remained without recurrence 1 year after surgery.

Fertility determinants after conservative surgery for mucinous borderline tumours of the ovary (excluding peritoneal pseudomyxoma)

BACKGROUND The aim of this study was to define determinants of fertility in patients treated conservatively for mucinous borderline ovarian tumours (MBOTs), and to compare outcomes after salpingo-oophorectomy or cystectomy. METHODS This was a retrospective cohort study of fertility results in a series of patients treated conservatively for MBOTs and desiring pregnancy. Conservative surgery was defined as preservation of the uterus and ovarian tissue in one or both adnexa(e). Fertility results were compared with patients who had undergone a cystectomy or a (salpingo-)oophorectomy. Only patients with a minimum of 1 year of follow-up were included. Epidemiological, surgical, histological parameters and other prognostic factors for fertility results were investigated. RESULTS A group of 31 patients who had been treated conservatively between 1997 and 2004 and who desired pregnancy were investigated. Patients were treated by unilateral salpingo-oophorectomy (USO) (n = 19) or cystectomy (n = 12). The 5-year recurrence-free survival rate was higher in the USO group compared with the cystectomy group (94.7 versus 49.1%, P = 0.041). Among the 31 women, 12 had become pregnant. The 5-year probabilities of pregnancy were comparable between the cystectomy and salpingo-oophorectomy groups (41.8 and 45.9%, respectively, P= 0.66). None of the other factors studied (epidemiological, surgical and histological parameters) were associated with fertility results. CONCLUSIONS The use of salpingo-oophorectomy rather than cystectomy should be preferred during conservative surgery for patients with MBOTs because it decreases the risk of recurrence and does not impair fertility.

Decreased type III collagen expression in human uterine cervix of prolapse uteri

The precise mechanism of prolapse uteri is not fully understood. There is evidence to suggest that abnormalities of collagen, the main component of extracellular matrix, or its repair mechanism, may predispose women to prolapse. To investigate the characteristic structure of human uterine cervix of patients with prolapse uteri, various types of collagen expression in the uterine cervix tissues of the prolapse uteri were compared to those of normal uterine cervix. After informed consent, 36 specimens of uterine cervical tissues were obtained at the time of surgery from 16 postmenopausal women with prolapse uteri (stage III-IV by the Pelvic Organ Prolapse Quantification examination) and 20 postmenopausal women without prolapse uteri (control group). Collagens were extracted from the uterine cervix tissues by salt precipitation methods. The relative levels of various collagens were evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The uterine cervix was longer in the patients with prolapse uteri than those of postmenopausal controls without prolapse uteri. The ratios of type III to type I collagen in the uterine cervical tissues were significantly decreased in the prolapse uteri, as compared to those of the postmenopausal uterine cervix without prolapse. These results suggest that decreased type III collagen expression may play an important role in determing the physiology and structure of the uterine cervix tissues of prolapse uteri.

Molecular cloning, expression and variation analyses of the dopamine D2 receptor gene in pig breeds in China

The dopamine D2 receptor (DRD2) is a crucial mediator for normal physiological processes. We cloned the pig DRD2 gene, investigated its distribution in tissues and identified polymorphisms by RT-PCR, quantitative real-time PCR and direct sequencing. Two Yorkshire pigs from Guangdong Academy of Agricultural Sciences (Guangzhou, China) were selected to clone the gene and investigate its expression; 16 individuals from four pig breeds (Yorkshire, Landrace, small-ear spotted, and Xinchang) were used to scan the variations. The two transcripts (DRD2L and DRD2S), obtained through insertion or deletion of exon 5 and part of 3'UTR, were found to encode 444- and 415-amino acid proteins, respectively. The 574-bp indel in 3'UTR comprises five miRNA targeting sites, based on bioinformatics predictions. The pig DRD2 gene expresses predominantly in the pituitary gland, and then in oviducts and the hypothalamus. Both DRD2L and DRD2S mRNA were detected in cerebrum, cerebellum, hypothalamus, pituitary gland, back muscle, oviduct, uterus, and testis tissues; DRD2L was more abundant than DRD2S. The DRD2 gene is located on chromosome 9 and contains seven exons. Sixty-one different sequences were identified in this gene; among seven in the coding region, only one altered the encoded amino acid. These findings will help us understand the functions of the DRD2 gene in pigs.

Abstract P4-06-18: Asporin Is Associated with Invasive Growth and Bone Metastasis of Breast Cancer

Abstract Background: In an earlier publication we identified asporin as a novel cancer related protein (BMC Cancer 2007, 7:55), also reported among bone metastasis specific genes (Cancer Letters 2009, 276:212-220). Asporin is a member of the class I small leucine-rich repeat proteoglycan family and besides affinity to collagen, it binds calcium and initiates collagen mineralization. Asporin is abundantly expressed in tissues with high remodeling rate (e.g. peridontal ligament) whereas extremely high turnover of matrix proteins is also associated with invasive cancer. Material and methods: Formalin-fixed paraffin-embedded tissues [invasive ductal (101) and lobular (39) carcinomas classified according to hormone receptors and Her-2; 5 bone metastases from autopsies] were immunohistochemically stained with in-house polyclonal peptide antibody against asporin. The antibody was validated on uterus tissue as a positive control together with a commercial antibody (Abcam). Data were evaluated by non-parametric Mann-Whitney U test and Spearman correlation coefficient. Expression of asporin in both tissues and cell lines was also queried in databases Gene Expression Omnibus, ArrayExpress and Oncomine. Expression of asporin was further tested by RT-PCR in selected breast cancer cell lines. Migration and invasion of MDA-MB-231 cells were tested using real-time cell analysis Xcelligence system (Roche). Transwells were coated by solidifying collagen for 6 hours (with or without recombinant asporin). Starved cells were seeded on the collagen in transwells while FBS was used as a chemoattractant in bottom wells. Results: Levels of asporin were significantly higher in invasive lobularcarcinomas than ductal ones (P&amp;lt;0.001). Asporin was also highly expressed in all bone metastases. Asporin expression positively correlated with tumor grade within the triple negative subgroup (Rs 0.4, P&amp;lt;0.05) but not in luminal and Her-2 subtypes. Expression of asporin in invasive breast carcinomas was also confirmed in other public datasets (e.g. Karnoub breast and Hess breast in Oncomine database). Surprisingly, we found no breast cancer cell line overexpressing asporin, which underlines the importance of the tumor microenvironment in vivo. MDA-MB-231 cells migrated faster through collagen I with recombinant asporin. Conclusion: Asporin is expressed by both invasive breast carcinomas and bone metastases. Mechanistically, asporin may affect extracellular matrix structure and promote invasive growth of breast cancer cells. Further investigation of the role of asporin in metastasizing to bone is additionally justified given its affinity for calcium. Acknowledgements This work was supported by grants NS 9956-4 from the Czech Ministry of Health and MSM 6198959216 from the Czech Ministry of Education. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-06-18.

Effect of carbon disulfide on expression of matrix metalloproteinase-2 and metalloproteinase-9 in embryo and uterus of pregnant mice

To observe the effect of carbon disulfide (CS(2)) on the expression of matrix metalloproteinase (MMP)-2, MMP-9 in mouse embryo and uterus tissues and to explore the mechanism of embryo toxicity induced by CS(2). At the phases of follicular development and embryonic implantation which was subdivided into early-implantation phase and late-implantation phase, mice were intraperitoneally exposed to CS(2) (the dosage was 631.4 mg/kg, and the volume was 0.1ml/10 g body weight) for 2 consecutive days. All indicators were got at the ninth day in gestation, and the expression of MMP-2 and MMP-9 in embryo and uterus tissues was analyzed by gelatin zymography. The number of implanted embryos significantly decreased after exposure at late-implantation phase (16.000 ± 12.166) compared with those of the control (30.700 ± 5.599, P < 0.05). Expression of MMP-2 and MMP-9 in embryos declined obviously at the three reproductive phases (P < 0.01), and the levels of MMP-2 and MMP-9 expression in embryos at the phases of late-implantation phase (0.6837 ± 0.0929, 0.7309 ± 0.0822) and follicular development (0.6222 ± 0.0997, 0.7520 ± 0.1068) were much lower than those of the control (1.0000 ± 0.0710, 1.0000 ± 0.0413, P < 0.01). Expression of MMP-2 and MMP-9 in uterus significantly increased at the phase of late-implantation (1.3153 ± 0.3032, 5.0210 ± 4.0307) compared with those of the control (1.0000 ± 0.1771, 1.0000 ± 0.0996, P < 0.01). Embryo toxicity of CS(2) is more obvious at the phase of late-implantation. Exposure to CS(2) disturbs expression of MMP-2 and MMP-9 in embryo and uterus tissues, which might be one of the important factors contributed to embryo toxicity induced by CS(2).

Genomic structure and characterization of mRNA expression pattern of porcine interferon gamma receptor 1 gene

Interferon gamma receptor (IFNGR) plays an important role in the biological effects of IFN-γ. In this study, porcine IFNGR1 cDNA was cloned and two transcripts both having a coding region of 1413 bp were identified. Porcine IFNGR1 cDNA shares 62.95%, 63.73%, 72.90% and 81.10% identity in nucleotide sequence; and 45.64%, 46.69%, 58.04% and 72.55% homology in amino acid sequence to those of rat, mouse, human and cattle, respectively. The porcine IFNGR1 genomic structure consists of seven exons and six introns and is located on porcine chromosome 1. The mRNA expression of porcine IFNGR1 gene is detected in all tissues examined, with strong expression in spleen and liver tissues and weak expression in cerebrum, cerebellum and uterus tissues, respectively. A different developmental pattern in IFNGR1 mRNA expression between Laiwu and Duroc breeds was revealed by real-time quantitative RT-PCR: in Duroc pigs, a significantly higher expression was found in the tissues of heart (P<0.05), liver (P<0.01), kidney (P<0.01) and skeletal muscle (P<0.05) of adult pigs compared to piglets. In porcine reproductive and respiratory syndrome virus (PRRSV)-infected Dapulian pigs, compared to the uninfected ones, the expression level of IFNGR1 mRNA in spleen was significantly up-regulated (P<0.05), whereas its expression in the lymph node was significantly down-regulated (P<0.05); in PRRSV-infected Duroc × Yorkshire × Landrace commercial pigs, however, the differences both in spleen and lymph node tissues were not significant.

Comparative embryology and mammalian cloning

A hypothesis has been advanced that logically combines "contradictory" facts concerning the early mammalian development and shows a natural relationship between the embryos developing from a fertilized ovum and from cells of the inner cell mass of blastocyst. When studying the theoretical questions of cloning, it is necessary to take into consideration the peculiarities of prenatal mammalian ontogenesis, which make themselves evident upon comparison with other animals. The absence of yolk in the mammalian ovum defines sharp differences in the early development between mammals and other Amniota. The whole asynchronic cleavage results in the formation of the morula followed by the blastocyst, which hatches from zona pellucida and is implanted into the uterus tissue. This fact allows us to consider the blastocyst as a mammalian larva, which is fed thanks to maternal organism. It is known that, in the body of a larva (blastocyst), a new embryo develops from some somatic cells. This process is known as a polyembryony, which is typical for the development of some parasitic insects. The polyembryony in turn is a variant of somatic embryogenesis, which is a form of asexual reproduction. Thus, two different embryos, "conceptus" and "embryo proper", have different origin: the first forms by the sexual way and the second, by the asexual. The investigation of the mechanisms of somatic embryogenesis in mammals will help us to find conditions necessary for the full reprograming of donor somatic nuclei and provide the successful development of reconstructed embryos.

Conservative Approach in Gynecologic Malignancies

Abstract Objective: This review was undertaken with the purpose of highlighting the options of a conservative approach in the management of gynecologic cancers so that clinicians exhibit some restraint before deciding on extensive surgeries as the management option. Background: Ovarian function and fertility are the main concerns while treating young women with gynecologic cancers. A conservative approach in management addresses these issues and aims at preserving the uterus and/or ovarian tissue. Early cervical and endometrial cancers, germ cell ovarian tumors, borderline ovarian tumors, and even early epithelial ovarian tumors are the gynecologic cancers where there is scope for a conservative approach. It may be in the form of radical trachelectomy for cervical cancer, medical (hormonal) treatment for endometrial carcinoma, or uterus/ovary-preserving surgeries for ovarian malignancies. On other end of the spectrum, there are advanced malignancies that may require extensive surgeries, which may further ...

Novel monoclonal antibodies detect Smad-interacting protein 1 (SIP1) in the cytoplasm of human cells from multiple tumor tissue arrays

Smad-interacting protein 1 (SIP1, also known as ZEB2) represses the transcription of E-cadherin and mediates epithelial–mesenchymal transition in development and tumor metastasis. Due to the lack of human SIP1-specific antibodies, its expression in human tumor tissues has not been studied in detail by immunohistochemistry. Hence, we generated two anti-SIP1 monoclonal antibodies, clones 1C6 and 6E5, with IgG1 and IgG2a isotypes, respectively. The specificity of these antibodies was shown by Western blotting studies using siRNA mediated downregulation of SIP1 and ZEB1 in a human osteosarcoma cell line. In the same context, we also compared them with 5 commercially available SIP1 antibodies. Antibody specificity was further verified in an inducible cell line system by immunofluorescence. By using both antibodies, we evaluated the tissue expression of SIP1 in paraffin-embedded tissue microarrays consisting of 22 normal and 101 tumoral tissues of kidney, colon, stomach, lung, esophagus, uterus, rectum, breast and liver. Interestingly, SIP1 predominantly displayed a cytoplasmic expression, while the nuclear localization of SIP1 was observed in only 6 cases. Strong expression of SIP1 was found in distal tubules of kidney, glandular epithelial cells of stomach and hepatocytes, implicating a co-expression of SIP1 and E-cadherin. Squamous epithelium of the esophagus and surface epithelium of colon and rectum were stained with moderate to weak intensity. Normal uterus, breast and lung tissues remained completely negative. By comparison with their normal tissues, we observed SIP1 overexpression in cancers of the kidney, breast, lung and uterus. However, SIP1 expression was found to be downregulated in tumors from colon, rectum, esophagus, liver and stomach tissues. Finally we did nuclear/cytoplasmic fractionation in 3 carcinoma cell lines and detected SIP1 in both fractions, nucleus being the dominant one. To our best knowledge, this is the first comprehensive immunohistochemical study of the expression of SIP1 in a series of human cancers. Our finding that SIP1 is not exclusively localized to nucleus suggests that the subcellular localization of SIP1 is regulated in normal and tumor tissues. These novel monoclonal antibodies may help elucidate the role of SIP1 in tumor development.