子宫内膜息肉研究进展

子宫内膜息肉(endometrial polyps,EMP)由子宫内膜腺体和含有厚壁血管的纤维化内膜间质构成,是局部子宫内膜过度增生形成的有蒂或无蒂的赘生物。子宫内膜息肉是最常见的子宫内膜病变之一,临床表现为子宫不规则出血,或月经量增多、不孕、绝经后出血等,也可无明显临床症状。子宫内膜息肉多数属良性病变,但其可恶变性已经被证实。子宫内膜息肉的发病机制目前尚不明确,传统观点认为其与慢性子宫内膜炎症有关,属慢性炎症范畴,即为生物致炎因子及长期反复机械性刺激所致的反应性增生物;近年来随着分子生物学研究的深入,发现子宫内膜息肉的发生与可能与激素调控下增殖与凋亡失衡相关。近来随着宫腔镜检查技术的推广及激素补充治疗人数的增多,子宫内膜息肉的发病率及检出率逐渐增加,加之经刮宫或电切治疗后复发率非常高,因此,子宫内膜息肉越来越受到临床医生的重视,现将子宫内膜息肉的研究进展作一综述。     

干细胞相关因子在子宫内膜息肉中的表达

探讨ipo13、c-kit、CD146、bcl-2和bax在子宫内膜息肉(endometrial polyp,EP)和正常内膜组织中的表达差异及临床意义。收集本院2010年3-7月行宫腔镜手术取得的40例子宫内膜息肉(病例组)与40例正常内膜组织(对照组)。采用实时荧光定量PCR技术(RT-PCR)检测ipo13、c-kit、bcl-2和bax mRNA的表达;免疫组织化学S-P法检测ipo13、CD146、bcl-2和bax蛋白的表达。无论在月经周期的增生期或分泌期,EP中ipo13、c-kit、CD146和bax mRNA和蛋白的表达均低于同期正常子宫内膜组织,差异有统计学意义(P<0.05),bcl-2均比同期正常子宫内膜增加,差异有统计学意义(P<0.05)。子宫内膜干/祖细胞活性异常和子宫内膜凋亡减少与子宫内膜息肉发病有关,其中子宫内膜干/祖细胞活性异常使内膜凋亡减少可能是EP发病的主要因素。     

Spatial and temporal characterization of endometrial mesenchymal stem-like cells activity during the menstrual cycle

The human endometrium is a highly dynamic tissue with the ability to cyclically regenerate during the reproductive life. Endometrial mesenchymal stem-like cells (eMSCs) located throughout the endometrium have shown to functionally contribute to endometrial regeneration. In this study we examine whether the menstrual cycle stage and the location in the endometrial bilayer (superficial and deep portions of the endometrium) has an effect on stem cell activities of eMSCs (CD140b⁺CD146⁺ cells). Here we show the percentage and clonogenic ability of eMSCs were constant in the various stages of the menstrual cycle (menstrual, proliferative and secretory). However, eMSCs from the menstrual endometrium underwent significantly more rounds of self-renewal and enabled a greater total cell output than those from the secretory phase. Significantly more eMSCs were detected in the deeper portion of the endometrium compared to the superficial layer but their clonogenic and self-renewal activities remained similar. Our findings suggest that eMSCs are activated in the menstrual phase for the cyclical regeneration of the endometrium.     

Mesenchymal stem cells support expansion of in vitro irradiated CD34(+) cells in the presence of SCF, FLT3 ligand, TPO and IL3: potential application to autologous cell therapy in accidentally irradiated victims

Ex vivo expansion of residual autologous hematopoietic stem and progenitor cells collected from victims soon after accidental irradiation (autologous cell therapy) may represent an additional or alternative approach to cytokine therapy or allogeneic transplantation. Peripheral blood CD34+ cells could be a useful source of cells for this process provided that collection and ex vivo expansion of hematopoietic stem and progenitor cells could be optimized. Here we investigated whether mesenchymal stem cells could sustain culture of irradiated peripheral blood CD34+ cells. In vitro irradiated (4 Gy 60Co gamma rays) or nonirradiated mobilized peripheral blood CD34+ cells from baboons were cultured for 7 days in a serum-free medium supplemented with stem cell factor+thrombopoietin+interleukin 3+FLT3 ligand (50 ng/ml each) in the presence or absence of mesenchymal stem cells. In contrast to cultures without mesenchymal stem cells, irradiated CD34+ cells cultured with mesenchymal stem cells displayed cell amplification, i.e. CD34+ (4.9-fold), CD34++ (3.8-fold), CD34++/Thy-1+ (8.1-fold), CD41+ (12.4-fold) and MPO+ (50.6-fold), although at lower levels than in nonirradiated CD34+ cells. Fourteen times more clonogenic cells, especially BFU-E, were preserved when irradiated cells were cultured on mesenchymal stem cells. Moreover, we showed that the effect of mesenchymal stem cells is related mainly to the reduction of apoptosis and involves cell-cell contact rather than production of soluble factor(s). This experimental model suggests that mesenchymal stem cells could provide a crucial tool for autologous cell therapy applied to accidentally irradiated victims.     

Endometrial and cervical polyps in 22 baboons (Papio sp.), 5 cynomolgus macaques (Macaca fascicularis) and one marmoset (Callithrix jacchus)

Background  Endometrial and cervical polyps are masses of endometrium or cervical epithelium that bulge into the uterine or cervical lumen. The physiopathology and contributing factors of endometrial polyps development are still unknown.
Methods  Clinical and pathology records of 28 non-human primates with histologically confirmed endometrial and cervical polyps were reviewed. Twenty-one baboons with endometrial polyps were evaluated for age at diagnosis, body weight, menstrual cycle length, presence of endometriosis and adenomyosis and number of offspring, cesarean sections, and stillbirths.
Results  Endometrial polyps in baboons were associated with increased age, decreased menstrual cycle lengths, endometriosis, and decreased parity. No differences were found for weight, adenomyosis, or number of cesarean sections or stillbirths.
Conclusions  Baboons are a promising model for the study of endometrial polyps because of their similarity to humans in both the development of endometrial polyps and association of many of the same risk factors.     

Isolation of Stem-Like Cancer Cells in Primary Endometrial Cancer Using Cell Surface Markers CD133 and CXCR4

Endometrial cancer (EC) is the most common familiar gynecologic malignant tumor identified in the female reproductive system and has been increasing yearly. In this study, we will identify the surface markers and stem cell markers related with cancer stem cells (CSCs) of EC. Tissue samples were obtained from endometrial cancer patients during surgical procedures. Single cells were isolated from the tissues for culturing, transfection into nude mice, and histopathology analysis. RT-PCR demonstrated that the cultured cells strongly expressed stemness-related genes, such as c-Myc, Sox-2, Nanog, Oct 4A, ABCG2, BMI-1, CK-18, Nestin and β-actin. The expression of surface markers CD24, CD133, CD47, CD29, CD44, CXCR4, SSEA3 and SSEA4, CD24, and CD133 and chemokine markers such as CXCR4 were measured by flow cytometry. Then the double percentage of CD133+CXCR4+ cells constituted 7.2% and 9.3% in EC cells originated from two different patients, respectively. The CD133+CXCR4+ primary endometrial cancer cells grew faster, exhibited high expression of mRNA of stemness-related genes, produced more spheres, and had higher clonogenic ability than other subpopulations. They are also more resistant to anti-cancer drugs than other subpopulations. These findings indicate that CD133+CXCR4+ cells may possess some characteristics of CSCs in primary endometrial cancer. These cell surface markers may be useful for the development of drugs against CSC molecular targets or as a predictive marker for poor prognosis in primary endometrial cancer.     

Significance of cytologically normal endometrial cells in cervical smears from postmenopausal women

OBJECTIVE: To determine the significance of cytologically normal endometrial cells in cervicovaginal (CV) smears from postmenopausal women over age 55 years. STUDY DESIGN: From January 1995 to January 1998, 220 women had CV smears demonstrating cytologically normal endometrial cells. The menopausal status, hormone replacement therapy (HRT) and information related to subsequent CV smears and endometrial sampling within 12 months of the initial diagnosis was recorded. RESULTS: Eighty-one of the 220 cases (36.8%) had histologic sampling of the endometrium. Thirty-four of 81 (42%) showed no endometrial pathology. Endometrial pathology was identified in 28 of 81 (34%), of which 19 were endometrial polyps (23.4%), 4 were endometrial hyperplasia (4.9%), 4 were endometrial carcinoma (4.9%) and 1 was a leiomyoma (1.2%). Nineteen (23.4%) were insufficient for diagnosis. Ninety-one of 220 women were on HRT, and 129 were not. In the group without HRT, endometrial disease was identified in 22/51 (43%) cases as compared to 6/30 (20%) in the group with HRT (P < .001). Endometrial carcinoma was identified in three (5.8%) cases and one (3.3%) case without and with HRT, respectively. CONCLUSION: Although the finding of normal endometrial cells in Pap smears from postmenopausal women was without any clinical significance in the majority of women in this study, in a small number it was associated with endometrial hyperplasia and carcinoma. Women who were not on HRT had a higher incidence of endometrial pathology.     

Reconstruction of endometrium from human endometrial side population cell lines

Endometrial regeneration is mediated, at least in part, by the existence of a specialized somatic stem cell (SSC) population recently identified by several groups using the side population (SP) technique. We previously demonstrated that endometrial SP displays genotypic, phenotypic and the functional capability to develop human endometrium after subcutaneous injection in NOD-SCID mice. We have now established seven human endometrial SP (hESP) cell lines (ICE 1–7): four from the epithelial and three from the stromal fraction, respectively. SP cell lines were generated under hypoxic conditions based on their cloning efficiency ability, cultured for 12–15 passages (20 weeks) and cryopreserved. Cell lines displayed normal 46XX karyotype, intermediate telomerase activity pattern and expressed mRNAs encoding proteins that are considered characteristic of undifferentiated cells (Oct-4, GDF3, DNMT3B, Nanog, GABR3) and those of mesodermal origin (WT1, Cardiac Actin, Enolase, Globin, REN). Phenotype analysis corroborated their epithelial (CD9+) or stromal (vimentin+) cell origin and mesenchymal (CD90+, CD73+ and CD45−) attributes. Markers considered characteristic of ectoderm or endoderm were not detected. Cells did not express either estrogen receptor alpha (ERα) or progesterone receptor (PR). The hESP cell lines were able to differentiate in vitro into adipocytes and osteocytes, which confirmed their mesenchymal origin. Finally, we demonstrated their ability to generate human endometrium when transplanted beneath the renal capsule of NOD-SCID mice. These findings confirm that SP cells exhibit key features of human endometrial SSC and open up new possibilities for the understanding of gynecological disorders such as endometriosis or Asherman syndrome. Our cell lines can be a valuable model to investigate new targets for endometrium proliferation in endometriosis.     

Long-term label retaining cells localize to distinct regions within the female reproductive epithelium

The uterus is an extremely plastic organ that undergoes cyclical remodeling including endometrial regeneration during the menstrual cycle. Endometrial remodeling and regeneration also occur during pregnancy and following parturition, particularly in hemochorial implanting species. The mechanisms of endometrial regeneration are not well understood. Endometrial stem/progenitor cells are proposed to contribute to endometrial regeneration in both humans and mice. BrdU label retention has been used to identify potential stem/progenitor cells in mouse endometrium. However, methods are not available to isolate BrdU label-retaining cells (LRC) for functional analyses. Therefore, we employed a transgenic mouse model to identify H2B-GFP LRCs throughout the female reproductive tract with particular interest on the endometrium. We hypothesized that the female reproductive tract contains a population of long-term LRCs that persist even following pregnancy and endometrial regeneration. Endometrial cells were labeled (pulsed) either transplacentally/translactationally or peripubertally. When mice were pulsed transplacentally/translactationally, the label was not retained in the uterus. However, LRCs were concentrated to the distal oviduct and endocervical transition zone (TZ) following natural (i.e., pregnancy/parturition induced) and mechanically induced endometrial regeneration. LRCs in the distal oviduct and endocervical TZ expressed stem cell markers and did not express ERα or PGR, implying the undifferentiated phenotype of these cells. Oviduct and endocervical TZ LRCs did not proliferate during endometrial re-epithelialization, suggesting that they do not contribute to the endometrium in a stem/progenitor cell capacity. In contrast, when mice were pulsed peripubertally long-term LRCs were identified in the endometrial glandular compartment in mice as far out as 9 months post-pulse. These findings suggest that epithelial tissue of the female reproductive tract contains 3 distinct populations of epithelial cells that exhibit stem/progenitor cell qualities. Distinct stem/progenitor-like cells localize to the oviduct, endometrium, and cervix.     

Isolation of Oct4+, Nanog+ and SOX2− mesenchymal cells from peripheral blood of a diabetes mellitus patient

Diabetes mellitus is a chronic metabolic disorder that affects millions of people worldwide. The most common form is type 2 diabetes mellitus, which results in impaired beta cell function combined with insulin resistance in peripheral organs. One recently proposed treatment approach is the use of adult stem cells derived from bone marrow in autologous stem cell transplantation. Alternatively, peripheral blood can be obtained in a more non-invasive manner. In this study, we isolated and cultured mesenchymal cells (MCs) from the peripheral blood of a diabetes mellitus patient. The cultured cells were large and elongated and had an in vitro migratory capacity in the culture dish. They expressed embryonic stem cell pluripotency markers Nanog and Oct 4 as well as mesenchymal markers CD105 and CD13, and they lacked expression of hematopoietic marker CD45. These characteristics suggest that these cells have a mesenchymal phenotype similar to that obtained from bone marrow cells. The SOX2 gene was downregulated in both the peripheral blood cells and the isolated mesenchymal cell line, indicating a defective mechanism of SOX2 in diabetes mellitus. The overall results of study demonstrate that peripheral blood can be used as a source of MCs from diabetes mellitus patients for use in future regenerative stem cell therapy and that this particular model system may be useful to study the mechanism of diabetes mellitus involving downregulation of the SOX2 cascade.     

Characteristics of Human Amniotic Fluid Mesenchymal Stem Cells and Their Tropism to Human Ovarian Cancer

The mesenchymal stem cells (MSCs) derived from amniotic fluid (AF) have become an attractive stem cells source for cell-based therapy because they can be harvested at low cost and avoid ethical disputes. In human research, stem cells derived from AF gradually became a hot research direction for disease treatment, specifically for their plasticity, their reduced immunogenicity and their tumor tropism regardless of the tumor size, location and source. Our work aimed to obtain and characterize human amniotic fluid mesenchymal stem cells (AFMSCs) and detect their ovarian cancer tropsim in nude mice model. Ten milliliters of twenty independent amniotic fluid samples were collected from 16-20 week pregnant women who underwent amniocentesis for fetal genetic determination in routine prenatal diagnosis in the first affiliated hospital of Harbin medical university. We successfully isolated the AFMSCs from thirteen of twenty amniotic fluid samples. AFMSCs presented a fibroblastic-like morphology during the culture. Flow cytometry analyses showed that the cells were positive for specific stem cell markers CD73,CD90, CD105, CD166 and HLA-ABC (MHC class I), but negative for CD 45,CD40, CD34, CD14 and HLA-DR (MHC class II). RT-PCR results showed that the AFMSCs expressed stem cell marker OCT4. AFMSCs could differentiate into bone cells, fat cells and chondrocytes under certain conditions. AFMSCs had the high motility to migrate to ovarian cancer site but didn’t have the tumorigenicity. This study enhances the possibility of AFMSCs as drug carrier in human cell-based therapy. Meanwhile, the research emphasis in the future can also put in targeting therapy of ovarian cancer.     

Characterization of fibroblast-like cells from the rat olfactory bulb

The isolation and characterization of stem cells from an alternative tissue is a subject of intensive investigation. In the present study, we have focused on the characterization of fibroblastic cells in olfactory bulb tissue of the rat. To this end, 4-6 week old rats were killed and their olfactory bulb tissue was dissected out. Olfactory bulb derived fibroblast-like cells were recovered by adhesion to cell culture plastic. The plastic adherent cultivated cells were induced to differentiate along osteoblastic, adipogenic and chondrogenic lineages. Furthermore, the expression of some surface antigens was investigated. We obtained purified cells with spindle shaped morphology in primary culture, which differentiated into mesenchymal lineages. These cells expressed CD29 and CD90 (Thy1.1) surface antigens, but not CD31, CD34 and CD45. Our results indicate that fibroblast-like cells from the olfactory bulb are mesenchymal stem cells in nature. Taken together, our data suggest that olfactory bulb tissue may constitute a new source of mesenchymal stem cells and could be used for the treatment of injury.     

Decreased expression of survivin, estrogen and progesterone receptors in endometrial tissues after radiofrequency treatment of dysfunctional uterine bleeding

ABSTRACT: BACKGROUND: The purpose of the research is to study the histopathology and expression of survivin, estrogen and progesterone receptors (ER/PR) in the endometrium of patients with dysfunctional uterine bleeding (DUB) treated with adiofrequency endometrial ablation (REA). METHODS: A total of 98 DUB patients were enrolled in this case-control study. Among them, 66 underwent REA treatment and 32 optioned for hormone therapy as the control group. Immunohistochemical analysis for survivin, ER and PR expression was carried out on endometrial tissue samples collected before and 6 to 7 months after treatment for both groups. RESULTS: Both hormone and REA treatment ameliorated menstrual bleeding of DUB patients, with the latter showing a significantly higher effective rate. Endometrial surface tissue was replaced by fibrosis tissue in the REA treatment group. REA treatment also significantly reduced the expression of survivin, ER, and PR. Endometrial surface tissues collected from the hormonetreated control group neither showed any apparent morphological alteration nor in the expression of those receptors. CONCLUSIONS: REA treatment changed endometrial surface tissue type from gland rich to gland poor, and significantly decreased the expression of survivin, ER, and PR. This may be an important contributing mechanism for the long-term curative effect and prevention of DUB recurrence.     

Significance of histiocytes on otherwise-normal cervical smears from postmenopausal women. A retrospective study of 108 cases

OBJECTIVE: To evaluate the significance of histiocytes on normal cervical smears from postmenopausal women and correlate them with endometrial pathology. STUDY DESIGN: Histiocytes were classified into three types. The clinical history was obtained from cytologic and surgical reports. RESULTS: Among 108 cervical smears, 13 had large, foamy histiocytes (type A), 88 had histiocytes resembling superficial endometrial stromal cells (type B), and 7 had variably sized histiocytes alone or in association with inflammatory or multinucleated cells (type C). Endometrial pathology was identified in 13 patients (12.0%): 4/13 with type A histiocytes (2 endometrial adenocarcinomas, 2 endometrial polyps), 8/88 with type B histiocytes (8 endometrial polyps) and 1/7 with type C histiocytes (endometrial polyp). Among 70 patients with no clinical indications for endometrial sampling except for the presence of histiocytes, 4 demonstrated endometrial pathology (all endometrial polyps). In contrast, endometrial pathology was identified in 9/38 with clinical indications for endometrial sampling. Among the 13 patients with endometrial pathology, 9 had a significant clinical history (sensitivity of 69.2%), and 4 had histiocytes as the only indication for endometrial biopsy (sensitivity of 30.8%). CONCLUSION: A significant clinical history is more predictive of endometrial pathology and outweighs the significance of histiocytes as an indication for endometrial biopsy.     

Identification of label-retaining perivascular cells in a mouse model of endometrial decidualization, breakdown, and repair

The human endometrium is incredibly dynamic, undergoing monthly cycles of growth and regression during a woman's reproductive life. Endometrial repair at the cessation of menstruation is critical for reestablishment of a functional endometrium receptive for embryo implantation; however, little is understood about the mechanisms behind this rapid and highly efficient process. This study utilized a functional mouse model of endometrial breakdown and repair to assess changes in endometrial vasculature that accompany these dynamic processes. Given that adult endometrial stem/progenitor cells identified in human and mouse endometrium are likely contributors to the remarkable regenerative capacity of endometrium, we also assessed label-retaining cells (LRC) as candidate stromal stem/progenitor cells and examined their relationship with endometrial vasculature. Newborn mouse pups were pulse-labeled with bromodeoxyuridine (BrdU) and chased for 5 wk before decidualization, endometrial breakdown, and repair were induced by hormonal manipulation. Mean vessel density did not change significantly throughout breakdown and repair; however, significantly elevated endothelial cell proliferation was observed in decidual tissue. Stromal LRC were identified throughout breakdown and repair, with significantly fewer observed during endometrial repair than before decidualization. A significantly higher percentage of LRC were associated with vasculature during repair than before decidualization, and a proportion were undergoing proliferation, indicative of their functional capacity. This study is the first to examine the endometrial vasculature and candidate stromal stem/progenitor cells in a functional mouse model of endometrial breakdown and repair and provides functional evidence suggesting that perivascular LRC may contribute to endometrial stromal expansion during the extensive remodeling associated with this process.     

宫腔镜联合B超诊断异常子宫出血124例临床分析

目的：探讨宫腔镜联合B超在诊治异常子宫出血的临床价值。方法：对124例异常子宫出血的病例进行回顾性分析,所有患者作B超及宫腔镜检查,宫腔内切除物或刮出物均送病理检查。结果：124例患者经病理检查确诊为子宫内膜息肉84例（67．7％）,合并宫颈息肉10例;子宫内膜增生症16例（12．9％）,其中单纯性增生12例,复杂性增生4例;子宫内膜不典型增生1例（0．8％）;子宫粘膜下肌瘤12例（9．7％）;子宫内膜样腺癌6例（4．8％）;子宫内膜炎3例（2．4％）;胚物残留2例（1．6％）。结论：宫腔镜联合B超检查是诊断异常子宫出血最好的方法.